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Identifying tumor-induced leukocyte subsets and their derived circulating factors has been instrumental in understanding cancer as a systemic disease. Nevertheless, how primary tumor-induced non-leukocyte populations in distal organs contribute to systemic spread remains poorly defined. Here, we report one population of tumor-inducible, erythroblast-like cells (Ter-cells) deriving from megakaryocyte-erythroid progenitor cells with a unique Ter-119+CD45-CD71+ phenotype. Ter-cells are enriched in the enlarged spleen of hosts bearing advanced tumors and facilitate tumor progression by secreting neurotrophic factor artemin into the blood. Transforming growth factor ß (TGF-ß) and Smad3 activation are important in Ter-cell generation. In vivo blockade of Ter-cell-derived artemin inhibits hepatocellular carcinoma (HCC) growth, and artemin deficiency abolishes Ter-cells' tumor-promoting ability. We confirm the presence of splenic artemin-positive Ter-cells in human HCC patients and show that significantly elevated serum artemin correlates with poor prognosis. We propose that Ter-cells and the secreted artemin play important roles in cancer progression with prognostic and therapeutic implications.
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Progresión de la Enfermedad , Eritroblastos/citología , Proteínas del Tejido Nervioso/sangre , Bazo/citología , Factor de Crecimiento Transformador beta/metabolismo , Animales , Apoptosis , Carcinoma Hepatocelular/metabolismo , Movimiento Celular , Proliferación Celular , Transición Epitelial-Mesenquimal , Regulación Neoplásica de la Expresión Génica , Receptores del Factor Neurotrófico Derivado de la Línea Celular Glial/metabolismo , Células Hep G2 , Humanos , Antígenos Comunes de Leucocito/metabolismo , Leucocitos/citología , Neoplasias Hepáticas/metabolismo , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Invasividad Neoplásica/genética , Transducción de SeñalRESUMEN
DEAD-box (DDX) helicases are vital for the recognition of RNA and metabolism and are critical for the initiation of antiviral innate immunity. Modification of RNA is involved in many biological processes; however, its role in antiviral innate immunity has remained unclear. Here we found that nuclear DDX member DDX46 inhibited the production of type I interferons after viral infection. DDX46 bound Mavs, Traf3 and Traf6 transcripts (which encode signaling molecules involved in antiviral responses) via their conserved CCGGUU element. After viral infection, DDX46 recruited ALKBH5, an 'eraser' of the RNA modification N6-methyladenosine (m6A), via DDX46's DEAD helicase domain to demethylate those m6A-modified antiviral transcripts. It consequently enforced their retention in the nucleus and therefore prevented their translation and inhibited interferon production. DDX46 also suppressed antiviral innate immunity in vivo. Thus, DDX46 inhibits antiviral innate responses by entrapping selected antiviral transcripts in the nucleus by erasing their m6A modification, a modification normally required for export from the nucleus and translation.
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Adenina/análogos & derivados , ARN Helicasas DEAD-box/genética , ARN Helicasas DEAD-box/metabolismo , Inmunidad Innata/genética , Transcripción Genética , Adenina/metabolismo , Desmetilasa de ARN, Homólogo 5 de AlkB/genética , Desmetilasa de ARN, Homólogo 5 de AlkB/metabolismo , Animales , Sitios de Unión , Línea Celular , Núcleo Celular/metabolismo , Expresión Génica , Técnicas de Silenciamiento del Gen , Humanos , Interferón Tipo I/biosíntesis , Macrófagos/inmunología , Macrófagos/metabolismo , Ratones , Ratones Noqueados , Motivos de Nucleótidos , Unión Proteica , Transporte de ARN , ARN Mensajero/genética , ARN Mensajero/metabolismo , Estomatitis Vesicular/genética , Estomatitis Vesicular/inmunología , Estomatitis Vesicular/metabolismo , Vesiculovirus/fisiología , Replicación ViralRESUMEN
This corrects the article DOI: 10.1038/ni.3830.
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Three-dimensional (3D) organic-inorganic lead halide perovskites have emerged in the past few years as a promising material for low-cost, high-efficiency optoelectronic devices. Spurred by this recent interest, several subclasses of halide perovskites such as two-dimensional (2D) halide perovskites have begun to play a significant role in advancing the fundamental understanding of the structural, chemical, and physical properties of halide perovskites, which are technologically relevant. While the chemistry of these 2D materials is similar to that of the 3D halide perovskites, their layered structure with a hybrid organic-inorganic interface induces new emergent properties that can significantly or sometimes subtly be important. Synergistic properties can be realized in systems that combine different materials exhibiting different dimensionalities by exploiting their intrinsic compatibility. In many cases, the weaknesses of each material can be alleviated in heteroarchitectures. For example, 3D-2D halide perovskites can demonstrate novel behavior that neither material would be capable of separately. This review describes how the structural differences between 3D halide perovskites and 2D halide perovskites give rise to their disparate materials properties, discusses strategies for realizing mixed-dimensional systems of various architectures through solution-processing techniques, and presents a comprehensive outlook for the use of 3D-2D systems in solar cells. Finally, we investigate applications of 3D-2D systems beyond photovoltaics and offer our perspective on mixed-dimensional perovskite systems as semiconductor materials with unrivaled tunability, efficiency, and technologically relevant durability.
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BACKGROUND AND AIMS: Hepatocarcinogenesis goes through HCC progenitor cells (HcPCs) to fully established HCC, and the mechanisms driving the development of HcPCs are still largely unknown. APPROACH AND RESULTS: Proteomic analysis in nonaggregated hepatocytes and aggregates containing HcPCs from a diethylnitrosamine-induced HCC mouse model was screened using a quantitative mass spectrometry-based approach to elucidate the dysregulated proteins in HcPCs. The heterotrimeric G stimulating protein α subunit (GαS) protein level was significantly increased in liver cancer progenitor HcPCs, which promotes their response to oncogenic and proinflammatory cytokine IL-6 and drives premalignant HcPCs to fully established HCC. Mechanistically, GαS was located at the membrane inside of hepatocytes and acetylated at K28 by acetyltransferase lysine acetyltransferase 7 (KAT7) under IL-6 in HcPCs, causing the acyl protein thioesterase 1-mediated depalmitoylation of GαS and its cytoplasmic translocation, which were determined by GαS K28A mimicking deacetylation or K28Q mimicking acetylation mutant mice and hepatic Kat7 knockout mouse. Then, cytoplasmic acetylated GαS associated with signal transducer and activator of transcription 3 (STAT3) to impede its interaction with suppressor of cytokine signaling 3, thus promoting in a feedforward manner STAT3 phosphorylation and the response to IL-6 in HcPCs. Clinically, GαS, especially K28-acetylated GαS, was determined to be increased in human hepatic premalignant dysplastic nodules and positively correlated with the enhanced STAT3 phosphorylation, which were in accordance with the data obtained in mouse models. CONCLUSIONS: Malignant progression of HcPCs requires increased K28-acetylated and cytoplasm-translocated GαS, causing enhanced response to IL-6 and driving premalignant HcPCs to fully established HCC, which provides mechanistic insight and a potential target for preventing hepatocarcinogenesis.
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Carcinoma Hepatocelular , Neoplasias Hepáticas , Lisina Acetiltransferasas , Humanos , Ratones , Animales , Neoplasias Hepáticas/patología , Carcinoma Hepatocelular/patología , Interleucina-6/metabolismo , Proteómica , Citoplasma/metabolismo , Proteínas de Unión al GTP/metabolismo , Lisina Acetiltransferasas/metabolismo , Factor de Transcripción STAT3/metabolismo , Histona Acetiltransferasas/metabolismoRESUMEN
The SARS-CoV-2 Omicron variant is characterized by its high transmissibility, which has caused a worldwide epidemiological event. Yet, it turns ominous once the disease progression degenerates into severe pneumonia and sepsis, presenting a horrendous lethality. To elucidate the alveolar immune or inflammatory landscapes of Omicron critical-ill patients, we performed single-cell RNA-sequencing (scRNA-seq) of bronchoalveolar lavage fluid (BALF) from the patients with critical pneumonia caused by Omicron infection, and analyzed the correlation between the clinical severity scores and different immune cell subpopulations. In the BALF of Omicron critical patients, the alveolar violent myeloid inflammatory environment was determined. ISG15+ neutrophils and CXCL10+ macrophages, both expressed the interferon-stimulated genes (ISGs), were negatively correlated with clinical pulmonary infection score, while septic CST7+ neutrophils and inflammatory VCAN+ macrophages were positively correlated with sequential organ failure assessment. The percentages of ISG15+ neutrophils were associated with more protective alveolar epithelial cells, and may reshape CD4+ T cells to the exhaustive phenotype, thus preventing immune injuries. The CXCL10+ macrophages may promote plasmablast/plasma cell survival and activation as well as the production of specific antibodies. As compared to the previous BALF scRNA-seq data from SARS-CoV-2 wild-type/Alpha critical patients, the subsets of neutrophils and macrophages with pro-inflammatory and immunoregulatory features presented obvious distinctions, suggesting an immune disparity in Omicron variants. Overall, this study provides a BALF single-cell atlas of Omicron critical patients, and suggests that alveolar interferon-responsive neutrophils and macrophages may extricate SARS-CoV-2 Omicron critical patients from the nasty fate of sepsis.
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Líquido del Lavado Bronquioalveolar , COVID-19 , Macrófagos , Neutrófilos , SARS-CoV-2 , Sepsis , Humanos , COVID-19/inmunología , COVID-19/virología , Neutrófilos/inmunología , Sepsis/inmunología , Sepsis/virología , SARS-CoV-2/inmunología , Masculino , Macrófagos/inmunología , Macrófagos/virología , Femenino , Persona de Mediana Edad , Líquido del Lavado Bronquioalveolar/virología , Líquido del Lavado Bronquioalveolar/inmunología , Líquido del Lavado Bronquioalveolar/citología , Anciano , Citocinas/inmunología , Interferones , Enfermedad Crítica , AdultoRESUMEN
Freespace optical (FSO) communication in an outdoor setting is complicated by atmospheric turbulence (AT). A time-varying (TV) multiplexed orbital angular momentum (OAM) propagation model to consider AT under transverse-wind conditions is formulated for the first time, and optimized dynamic correction periods for various TV AT situations are found to improve the transmission efficiency. The TV nature of AT has until now been neglected from modeling of OAM propagation models, but it is shown to be important. First, according to the Taylor frozen-turbulence hypothesis, a series of AT phase screens influenced by transverse wind are introduced into the conventional angular-spectrum propagation analysis method to model both the temporal and spatial propagation characteristics of multiplexed OAM beams. Our model shows that while in weak TV AT, the power standard deviation of lower-order modes is usually smaller than that of higher-order modes, the phenomena in strong TV AT are qualitatively different. Moreover, after analyzing the effective time of each OAM phase correction, optimized dynamic correction periods for a dynamic feedback communication link are obtained. An optimized result shows that, under the moderate TV AT, both a system BER within the forward-error-correction limit and a low iterative computation volume with 6% of the real-time correction could be achieved with a correction period of 0.18 s. The research emphasizes the significance of establishing a TV propagation model for exploring the effect of TV AT on multiplexed OAM beams and proposing an optimized phase-correction mechanism to mitigate performance degradation caused by TV AT, ultimately enhancing overall transmission efficiency.
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Titanium-oxo cluster (TOC)-based metal-organic frameworks (MOFs) have received considerable attention in recent years due to their ability to expand the application of TOCs to fields that require highly stable frameworks. Herein, a new cyclic TOC formulated as [Ti6O6(OiPr)8(TTFTC)(phen)2]2 (1, where TTFTC = tetrathiafulvalene tetracarboxylate and phen = phenanthroline) was crystallographically characterized. TOC 1 takes a rectangular ring structure with two phen-modified Ti6 clusters as the width and two TTFTC ligands as the length. An intracluster ligand-to-ligand (TTF-to-phen) charge transfer in 1 was found for TOCs for the first time. Compound 1 undergoes topotactic conversion to generate stable TOC-MOF P1, in which the rectangular framework in 1 formed by a TOC core and ligands is retained, as verified by comprehensive characterization. P1 shows an efficient and rapid selective adsorption capacity for cationic dyes. The experimental adsorption capacity (qex) of P1 reaches a value of up to 789.2 mg/g at 298 K for the crystal violet dye, which is the highest among those of various adsorbents. The calculated models are first used to reveal the structure-property relationship of the cyclic host to different guest dyes. The results further confirmed the host MOF structure of P1.
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The oleaginous yeast Yarrowia lipolytica represents a potential microbial cell factory for the recombinant production of various valuable products. Currently, the commonly used selection markers for transformation in Y. lipolytica are limited, and successive genetic manipulations are often restricted by the number of available selection markers. In our study, we developed a dominant marker, dsdA, which encodes a D-serine deaminase for genetic manipulation in Y. lipolytica. In Y. lipolytica, this marker confers the ability to use D-serine as a nitrogen source. In addition, the selection conditions of several infrequently used dominant markers including bleoR (zeocin resistance), kanMX (G418 resistance), and guaB (mycophenolic acid resistance) were also analyzed. Our results demonstrated that these selection markers can be used for the genetic manipulation of Y. lipolytica and their selection conditions were different for various strains. Ultimately, the selection markers tested here will be useful to expand the genetic toolbox of Y. lipolytica. KEY POINTS: ⢠The dsdA from Escherichia coli was developed as a dominant marker. ⢠The applicability of several resistance markers in Y. lipolytica was determined. ⢠We introduced the Cre/mutant lox system for marker recycling.
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Yarrowia , Yarrowia/genética , Marcadores Genéticos/genéticaRESUMEN
Saccharomyces cerevisiae is an important model eukaryotic microorganism and widely applied in fundamental research and the production of various chemicals. Its ability to efficiently and precisely control the expression of multiple genes is valuable for metabolic engineering. The clustered regularly interspaced short palindromic repeats (CRISPR)-mediated regulation enables complex gene expression programming; however, the regulation efficiency is often limited by the efficiency of pertinent regulators. Here, we developed CRISPR-mediated protein-tagging signal amplification system for simultaneous multiplexed gene activation and repression in S. cerevisiae. By introducing protein scaffolds (SPY and SunTag systems) to recruit multiple copies of regulators to different nuclease-deficient CRISPR proteins and design optimization, our system amplified gene regulation efficiency significantly. The gene activation and repression efficiencies reached as high as 34.9-fold and 95%, respectively, being 3.8- and 8.6-fold higher than those observed on the direct fusion of regulators with nuclease-deficient CRISPR proteins, respectively. We then applied the orthogonal bifunctional CRISPR-mediated transcriptional regulation system to regulate the expression of genes associated with 3-hydroxypropanoic acid production to deduce that CRISPR-associated regulator recruiting systems represent a robust method for simultaneously regulating multiple genes and rewiring metabolic pathways.
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Sistemas CRISPR-Cas , Ingeniería Metabólica , Saccharomyces cerevisiae , Activación Transcripcional , Repeticiones Palindrómicas Cortas Agrupadas y Regularmente Espaciadas , Endonucleasas/metabolismo , Edición Génica/métodos , Ácido Láctico/análogos & derivados , Ácido Láctico/biosíntesis , Ingeniería Metabólica/métodos , Saccharomyces cerevisiae/genéticaRESUMEN
DNA G4-structures from human c-MYC promoter and telomere are considered as important drug targets; however, the developing of small-molecule-based fluorescent binding ligands that are highly selective in targeting these G4-structures over other types of nucleic acids is challenging. We herein report a new approach of designing small molecules based on a non-selective thiazole orange scaffold to provide two-directional and multi-site interactions with flanking residues and loops of the G4-motif for better selectivity. The ligands are designed to establish multi-site interactions in the G4-binding pocket. This structural feature may render the molecules higher selectivity toward c-MYC G4s than other structures. The ligand-G4 interaction studied with 1H NMR may suggest a stacking interaction with the terminal G-tetrad. Moreover, the intracellular co-localization study with BG4 and cellular competition experiments with BRACO-19 may suggest that the binding targets of the ligands in cells are most probably G4-structures. Furthermore, the ligands that either preferentially bind to c-MYC promoter or telomeric G4s are able to downregulate markedly the c-MYC and hTERT gene expression in MCF-7 cells, and induce senescence and DNA damage to cancer cells. The in vivo antitumor activity of the ligands in MCF-7 tumor-bearing mice is also demonstrated.
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Antineoplásicos/química , Neoplasias de la Mama , G-Cuádruplex , Animales , Neoplasias de la Mama/tratamiento farmacológico , Neoplasias de la Mama/genética , Diseño de Fármacos , Femenino , Genes myc , Humanos , Ligandos , Células MCF-7 , Ratones , Regiones Promotoras Genéticas , TelómeroRESUMEN
AIM: Guanylate-binding protein 5 (GBP5) is an interferon (IFN)-inducible GTPase that plays a crucial role in the cell-autonomous immune response against microbial infections. In this study, we investigated the immunoregulatory role of GBP5 in the pathogenesis of dental pulpitis. METHODOLOGY: Gene-set enrichment analysis (GSEA) was utilized to evaluate the IFN-γ signalling pathway, and the differential expression of GBP mRNA in normal versus inflamed dental pulp tissues was screened, based on Gene Expression Omnibus (GEO) datasets associated with pulpitis. Both normal pulp tissues and inflamed pulp tissues were used for experiments. The expression of IFNs and GBPs was determined by qRT-PCR. Immunoblotting and double immunofluorescence were performed to examine the cellular localization of GBP5 in dental pulp tissues. For the functional studies, IFN-γ priming or lentivirus vector-delivered shRNA was used to, respectively, overexpress or knock down endogenous GBP5 expression in human dental pulp stem cells (HDPSCs). Subsequently, LPS was used to stimulate HDPSCs (overexpressing or with knocked-down GBP5) to establish an in vitro model of inflammation. qRT-PCR and ELISA were employed to examine the expression of proinflammatory cytokines (IL-6, IL-8 and IL-1ß) and cyclooxygenase 2 (COX2). Every experiment has three times of biological replicates and three technical replicates, respectively. Statistical analysis was performed using the Student's t-test and one-way ANOVA, and a p-value of <.05 was considered statistically significant. RESULTS: GSEA analysis based on the GEO dataset revealed a significant activation of the IFN-γ signalling pathway in the human pulpitis group. Among the human GBPs evaluated, GBP5 was selectively upregulated in inflamed dental pulp tissues and predominantly expressed in dental pulp cells. In vitro experiments demonstrated that IFN-γ robustly induced the expression of GBP5 in HDPSCs. Knockdown of GBP5 expression in HDPSCs significantly amplified the LPS-induced upregulation of inflammatory mediators (IL-6, IL-8, IL-1ß and COX2) both with and without IFN-γ priming. CONCLUSION: Our findings demonstrated that GBP5 partook in the pathogenesis of dental pulpitis. The involvement of GBP5 in pulpitis appeared to coordinate the regulation of inflammatory cytokines. Knockdown of GBP5 contributed to the exacerbation of LPS-mediated inflammation.
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Pulpitis , Humanos , Ciclooxigenasa 2/metabolismo , Citocinas/metabolismo , Pulpa Dental , Inflamación/metabolismo , Interleucina-6/metabolismo , Interleucina-8/metabolismo , Lipopolisacáridos/farmacología , Lipopolisacáridos/metabolismo , Pulpitis/metabolismoRESUMEN
AIM: Dental pulp is richly innervated by nerve fibres, which are mainly involved in the sensation of pain. Aside from pain sensation, little is known regarding the role of dental innervation in reparative dentine formation. We herein generated a mouse model of experimental dentine injury to examine nerve sprouting within the odontoblast and subodontoblastic layers and investigated the potential effects of this innervation in reparative dentinogenesis. METHODOLOGY: Mouse tooth cavity model (bur preparation + etching) was established, and then nerve sprouting, angiogenesis and reparative dentinogenesis were determined by histological and immunofluorescent staining at 1, 3, 7, 14 and 28 days postoperatively. We also established the mouse-denervated molar models to determine the role of sensory and sympathetic nerves in reparative dentinogenesis, respectively. Finally, we applied calcitonin gene-related peptide (CGRP) receptor antagonist to analyse the changes in angiogenesis and reparative dentinogenesis. RESULTS: Sequential histological results from dentine-exposed teeth revealed a significant increase in innervation directly beneath the injured area on the first day after dentine exposure, followed by vascularisation and reparative dentine production at 3 and 7 days, respectively. Intriguingly, abundant type H vessels (CD31+ Endomucin+ ) were present in the innervated area, and their formation precedes the onset of reparative dentine formation. Additionally, we found that sensory denervation led to blunted angiogenesis and impaired dentinogenesis, while sympathetic denervation did not affect dentinogenesis. Moreover, a marked increase in the density of CGRP+ nerve fibres was seen on day 3, which was reduced but remained elevated over the baseline level on day 14, whereas the density of substance P-positive nerve fibres did not change significantly. CGRP receptor antagonist-treated mice showed similar results as those with sensory denervation, including impairments in type H angiogenesis, which confirms the importance of CGRP in the formation of type H vessels. CONCLUSIONS: Dental pulp sensory nerves act as an essential upstream mediator to promote angiogenesis, including the formation of type H vessels, and reparative dentinogenesis. CGRP signalling governs the nerve-vessel-reparative dentine network, which is mostly produced by newly dense sensory nerve fibres within the dental pulp.
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Péptido Relacionado con Gen de Calcitonina , Dentina Secundaria , Ratones , Animales , Pulpa Dental/inervación , Angiogénesis , DolorRESUMEN
The in situ self-catalyzed growth of manganese-embedded 3D flakes-coated carbon rods (GFC) as an efficient oxygen-reduction reaction (ORR) catalyst of Zinc-air batteries is described for the first time. By optimizing the amount of Mn in the precursor, a series of 3D graphene-like flakes-coated carbon rods were synthesized. GFC with a doping amount of Mn of 10 % (GFC-10) exhibits excellent ORR performance with an onset potential of 0.94â V (vs. reversible hydrogen electrode). The Zinc-air battery is constructed with GFC-10 as the cathode catalyst, and it exhibits a peak power density of 128.9â mW cm-2 and a cycling stability of 75â h at a current density of 10â mA cm-2 , which are superior to the commercial 20â wt% Pt/C-based Zinc-air battery. Interestingly, the introduction of Mn facilitates the self-catalyzed growth of carbon rods, and the change of Mn amount can effectively regulate the morphology of materials. The improved ORR performance of the catalyst is ascribed to the synergistic effect of unique hierarchical porous structure, high-charge transport capacity, abundant carbon defects/edges and Mn-Nx sites. This research provides a new avenue to fabricating highly active Mn-based electrocatalysts for renewable energy systems.
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Grafito , Manganeso , Humanos , Carbono , Zinc , Hipoxia , OxígenoRESUMEN
Twisted moiré photonic crystal is an optical analog of twisted graphene or twisted transition metal dichalcogenide bilayers. In this paper, we report the fabrication of twisted moiré photonic crystals and randomized moiré photonic crystals and their use in enhanced extraction of light in light-emitting diodes (LEDs). Fractional diffraction orders from randomized moiré photonic crystals are more uniform than those from moiré photonic crystals. Extraction efficiencies of 76.5%, 77.8% and 79.5% into glass substrate are predicted in simulations of LED patterned with twisted moiré photonic crystals, defect-containing photonic crystals and random moiré photonic crystals, respectively, at 584 nm. Extraction efficiencies of optically pumped LEDs with 2D perovskite (BA)2(MA)n-1PbnI3n+1ofn= 3 and (5-(2'-pyridyl)-tetrazolato)(3-CF3-5-(2'-pyridyl)pyrazolato) platinum(II) (PtD) have been measured.
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AIM: Dentinal tubules serve as disease-causing channels for infiltration and penetration of bacteria and their by-products; which are regarded as the major driver of pathogenesis in pulpal inflammation and infection. In this study, we aimed to evaluate the transdentinal potential of nanoscale cetylpyridinium chloride/cholesterol (CPC/Chol) sterosomes, which are a recently developed type of cationic non-phospholipid liposomal nanocarrier; as well as their intrinsic and universal antibacterial activity. METHODOLOGY: Cetylpyridinium chloride/cholesterol sterosomes were formulated, with a hydrodynamic diameter of 134 ± 4 nm, a low polydisperse index of 0.161 ± 0.007, and a positive zeta potential of 41 ± 3 mV at pH 7.4. Transdentinal diffusion ability of sterosomes was evaluated using human dentine blocks in vitro, and Wistar rat molar teeth in vivo. The intrinsic antibacterial activities of CPC/Chol sterosomes against Enterococcus faecalis, Streptococcus mutans, Fusobacterium nucleatum, and Porphyromonas gingivalis were further examined. RESULTS: Cetylpyridinium chloride/cholesterol sterosomes successfully penetrated through the dentinal tubules, and diffused into the pulp, which could be internalized by dental pulp cells with a high efficiency. In addition, they exhibited substantial levels of intrinsic antibacterial activity against these Gram-positive and Gram-negative endodontic bacteria and their biofilms. CONCLUSIONS: Given its high penetration and diffusion ability through the dentine and pulp, great potential for multi-drug delivery, and distinct intrinsic antibacterial activity; sterosome-based nanocarriers might serve as a promising therapeutic strategy aimed at targeting various specific pathways associated with pulpal diseases. This will help determine and characterize the most appropriate prophylactic and therapeutic targets for early intervention in our future dentistry practice.
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Cetilpiridinio , Liposomas , Animales , Ratas , Humanos , Cetilpiridinio/farmacología , Ratas Wistar , Colesterol , Antibacterianos/farmacologíaRESUMEN
The chemical components of Huanglian Decoction were identified by ultra-performance liquid chromatography-quadrupole-time-of-flight-tandem mass spectrometry(UPLC-Q-TOF-MS/MS) technology. The gradient elution was conducted in Agilent ZORBAX Extend-C_(18) column(2.1 mm×100 mm, 1.8 µm) with the mobile phase of 0.1% formic acid aqueous solution(A)-acetonitrile(B) at a flow rate of 0.3 mL·min~(-1) and the column temperature of 35 â. The MS adopted the positive and negative ion mode of electrospray ionization(ESI), and the MS data were collected under the scanning range of m/z 100-1 500. Through high-resolution MS data analysis, combined with literature comparison and confirmation of reference substances, this paper identified 134 chemical components in Huanglian Decoction, including 12 alkaloids, 23 flavonoids, 22 terpenes and saponins, 12 phenols, 7 coumarins, 12 amino acids, 23 organic acids, and 23 other compounds, and the medicinal sources of the compounds were ascribed. Based on the previous studies, 7 components were selected as the index components. Combined with the network pharmacology research and analysis me-thods, the protein and protein interaction(PPI) network information of the intersection targets was obtained through the STRING 11.0 database, and 20 core targets of efficacy were screened out. In this study, UPLC-Q-TOF-MS/MS technology was successfully used to comprehensively analyze and identify the chemical components of Huanglian Decoction, and the core targets of its efficacy were discussed in combination with network pharmacology, which laid the foundation for clarifying the material basis and quality control of Huanglian Decoction.
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Medicamentos Herbarios Chinos , Espectrometría de Masas en Tándem , Farmacología en Red , Cromatografía Líquida de Alta Presión , Medicamentos Herbarios Chinos/química , TecnologíaRESUMEN
ß-Carotene is a provitamin A precursor and an important antioxidant that is used widely in the aquaculture, food, cosmetic, and pharmaceutical industries. Oleaginous Yarrowia lipolytica has been demonstrated as a competitive producer microorganism for the production of hydrophobic ß-carotene through rational engineering strategies. However, the limited understanding of the complexity of the metabolic network between carotenoid biosynthesis and other cellular processes has hampered further advancement. Genome-scale mutagenesis and high-throughput screening of mutagenesis libraries have been extensively employed in gene mining or in the identification of key targets associated with particular phenotypes. In this study, we developed a fluorescence-activated cell-sorting approach for the effective high-throughput screening of high-ß-carotene-producing strains. Using this approach, millions of mutants were screened rapidly, and new gene targets involved in lipid metabolism, sterol metabolism, signal transduction, and stress response were identified. The disruption of the genes affecting fatty acid oxidation, lipid composition, and sterol transcriptional regulation (4CL-8, GCS, and YIsterTF) increased ß-carotene significantly. By engineering these targets in a high-ß-carotene production, a strain that produced 9.4 g/L ß-carotene was constructed. Here, we used a flow cytometry approach to improve screening efficiency and eliminate the interference of intermediate metabolites. The targets obtained in this study can be used in studies focusing on metabolic engineering in the future for improving carotenoid production. IMPORTANCE ß-Carotene is a high-value-added product that is widely used in the aquaculture, food, cosmetic, and pharmaceutical industries. In our previous study, Yarrowia lipolytica has been engineered extensively to produce ß-carotene. To further improve its production, high-throughput screening and the identification of new beneficial gene targets are required. Herein, we developed a fluorescence-activated cell-sorting approach for the effective high-throughput screening of high-ß-carotene-producing strains. Using this approach, millions of mutants were screened rapidly, and new gene targets involved in lipid metabolism, sterol metabolism, signal transduction, and stress response were identified. The disruption of the genes affecting fatty acid oxidation, lipid composition, and sterol transcriptional regulation (4CL-8, GCS, and YIsterTF) increased ß-carotene significantly. By engineering these targets in a high-ß-carotene production, a strain that produced 9.4 g/L ß-carotene was constructed.
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Yarrowia , Antioxidantes/metabolismo , Ácidos Grasos/metabolismo , Citometría de Flujo , Ensayos Analíticos de Alto Rendimiento , Lípidos , Provitaminas/metabolismo , Esteroles/metabolismo , Yarrowia/genética , Yarrowia/metabolismo , beta Caroteno/metabolismoRESUMEN
Ultrahigh-order optical orbital angular momentum (OAM) states of the identification over ±270 orders are implemented by annular phase grating (APG) and Gaussian beams with different wavelengths. Particularly, the far-field diffraction intensity patterns feature the spiral stripes instead of Hermitian-Gaussian (HG)-like fringes. It's worth noting that the spiral stripes present uniform distribution, thus the order of OAM states can be intuitively acquired. More specifically, the OAM states can be confirmed from the total amount and rotating direction of the spiral stripes. Compared with traditional methods, the propose scheme contributes to the perfect-distributed and sharper spiral stripes. Moreover, it also makes an easier observation of the patterns in the CCD camera with limited imaging targets. In our experimental setup, the optical filter is removed and the APG parameters are not strictly required. Therefore, the propose optical transmission system is equipped with the advantages of efficiency, robustness and low cost, which paves a promising way for the communication capacity enhancement.