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Esterases are crucial for aryloxyphenoxypropionate herbicide (AOPP) biodegradation. However, the underlying molecular mechanisms of AOPP biodegradation by esterases are poorly understood. In the current work, Corynebacterium sp. Z-1 was isolated and found to degrade multiple AOPPs, including quizalofop-p-ethyl (QPE), haloxyfop-p-methyl (HPM), fenoxaprop-p-ethyl (FPE), cyhalofop-butyl (CYB), and clodinafop-propargyl (CFP). A novel esterase, QfeH, which catalyzes the cleavage of ester bonds in AOPPs to form AOPP acids, was identified from strain Z-1. The catalytic activities of QfeH toward AOPPs decreased in the following order: CFP > FPE > CYB > QPE > HPM. Molecular docking, computational analyses, and site-directed mutagenesis indicated the catalytic mechanisms of QfeH-mediated degradation of different AOPPs. Notably, the key residue S159 is essential for the activity of QfeH. Moreover, V222Y, T227M, T227A, A271R, and M275K mutants, exhibiting 2.9-5.0 times greater activity than QfeH, were constructed. This study facilitates the mechanistic understanding of AOPPs bioremediation by esterases.
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Biodegradación Ambiental , Corynebacterium , Esterasas , Herbicidas , Herbicidas/metabolismo , Herbicidas/química , Esterasas/metabolismo , Esterasas/genética , Esterasas/química , Corynebacterium/metabolismo , Corynebacterium/genética , Corynebacterium/enzimología , Proteínas Bacterianas/metabolismo , Proteínas Bacterianas/genética , Proteínas Bacterianas/química , Simulación del Acoplamiento Molecular , Propionatos/metabolismoRESUMEN
Methanol is not only a promising liquid hydrogen carrier but also an important feedstock chemical for chemical synthesis. Catalyst design is vital for enabling the reactions to occur under ambient conditions. This study reports a new class of van der Waals heterojunction photocatalyst, which is synthesized by hot-injection method, whereby carbon dots (CDs) are grown in situ on ZnSe nanoplatelets (NPLs), i.e., metal chalcogenide quantum wells. The resultant organic-inorganic hybrid nanoparticles, CD-NPLs, are able to perform methanol dehydrogenation through CH splitting. The heterostructure has enabled light-induced charge transfer from the CDs into the NPLs occurring on a sub-nanosecond timescale, with charges remaining separated across the CD-NPLs heterostructure for longer than 500 ns. This resulted in significantly heightened H2 production rate of 107 µmole·g-1·h-1 and enhanced photocurrent density up to 34 µA cm-2 at 1 V bias potential. EPR and NMR analyses confirmed the occurrence of α-CH splitting and CC coupling. The novel CD-based organic-inorganic semiconductor heterojunction is poised to enable the discovery of a host of new nano-hybrid photocatalysts with full tunability in the band structure, charge transfer, and divergent surface chemistry for guiding photoredox pathways and accelerating reaction rates.
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Constructing I single-atom (ISA) doped CoP electrocatalyst for HER is extremely challenging and has not been reported to date. Herein, an ISA doping-phosphatization strategy is proposed to prepare a novel I single-atom doped P-rich CoPn nanocluster@CoP electrocatalyst (ISA-CoPn/CoP) with enhanced HER performance first. ISA-CoPn/CoP shows a low overpotential of only 44 and 81 mV in 0.5 m H2SO4 solution, to drive a current density of 10 and 100 mA cm-2. ISA and P-rich CoPn nanocluster show unique synergies, which can optimize the H adsorption energy and accelerate the kinetics of HER in the CoP system. The intermediate IâH bond vibration peak is directly observed through in situ Raman testing, demonstrating that ISA doping helps accelerate the HER process. Additionally, the ΔGH of ISA-CoPn/CoP is only 0.05 eV by density functional theory (DFT) calculation, which is conducive to H2 evolution.
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Few-shot class-incremental learning (FSCIL) aims to continually learn novel data with limited samples. One of the major challenges is the catastrophic forgetting problem of old knowledge while training the model on new data. To alleviate this problem, recent state-of-the-art methods adopt a well-trained static network with fixed parameters at incremental learning stages to maintain old knowledge. These methods suffer from the poor adaptation of the old model with new knowledge. In this work, a dynamic clustering and recovering network (DyCR) is proposed to tackle the adaptation problem and effectively mitigate the forgetting phenomena on FSCIL tasks. Unlike static FSCIL methods, the proposed DyCR network is dynamic and trainable during the incremental learning stages, which makes the network capable of learning new features and better adapting to novel data. To address the forgetting problem and improve the model performance, a novel orthogonal decomposition mechanism is developed to split the feature embeddings into context and category information. The context part is preserved and utilized to recover old class features in future incremental learning stages, which can mitigate the forgetting problem with a much smaller size of data than saving the raw exemplars. The category part is used to optimize the feature embedding space by moving different classes of samples far apart and squeezing the sample distances within the same classes during the training stage. Experiments show that the DyCR network outperforms existing methods on four benchmark datasets. The code is available at: https://github.com/zichengpan/DyCR.
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BACKGROUND: The Talos stent-graft has extended length to improve aortic remodeling, and distal porous design to decrease the rate of spinal cord ischemia (SCI). This study retrospectively analyzed its mid-term outcomes for uncomplicated type B aortic dissection in a multicenter study. METHODS: The primary safety end point was 30-day major adverse events, including all-cause mortality, dissection-related mortality, conversion to open surgery, and device-related adverse events. The primary efficacy end point was treatment success at 12 months postoperation, defined as no technical failure or secondary dissection-related reintervention. The survival status of the patients was visualized using the Kaplan-Meier curve. Aortic growth was assessed at 4 levels, and SCI was evaluated at 12 months. RESULTS: 113 patients participated with a mean age of 54.4 (11.1) years and 71.7% (81/113) were male. The 30-day mortality was 0.9% (1/113), no conversions to open surgery or device-related adverse events were recorded. The 12-month treatment success rate was 99.1% (112/113), with no dissection-related reinterventions. There was no spinal cord or visceral ischemia at 12 months. At a median of 34 months follow-up, 9 further deaths were recorded and the 3-year survival rate was 91.7%. The percentage of aortic growth was 1.8% (2/111) at the tracheal bifurcation, 3.6% (4/111) below the left atrium, 6.0% (5/83) above the celiac artery, and 12.1% (9/74) below the lower renal artery. The total thrombosis rate of the false lumen at the stented segment was 80.5% (91/113). CONCLUSIONS: The results showed satisfactory results of Talos stent-graft in terms of safety and efficacy. More data are needed to confirm the long-term performance.
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Disección Aórtica , Implantación de Prótesis Vascular , Prótesis Vascular , Procedimientos Endovasculares , Diseño de Prótesis , Stents , Humanos , Masculino , Persona de Mediana Edad , Femenino , Disección Aórtica/cirugía , Disección Aórtica/diagnóstico por imagen , Disección Aórtica/mortalidad , Estudios Retrospectivos , Implantación de Prótesis Vascular/instrumentación , Implantación de Prótesis Vascular/efectos adversos , Implantación de Prótesis Vascular/mortalidad , Resultado del Tratamiento , Factores de Tiempo , Procedimientos Endovasculares/instrumentación , Procedimientos Endovasculares/efectos adversos , Procedimientos Endovasculares/mortalidad , Adulto , Anciano , Factores de Riesgo , Porosidad , Aneurisma de la Aorta/cirugía , Aneurisma de la Aorta/diagnóstico por imagen , Aneurisma de la Aorta/mortalidad , Complicaciones Posoperatorias/etiología , JapónRESUMEN
OBJECTIVES: The purpose of this study was to investigate the prognostic significance of radiomics in conjunction with hematological parameters in relation to the overall survival (OS) of individuals diagnosed with esophageal squamous cell carcinoma (ESCC) following definitive chemoradiotherapy (dCRT). METHODS: In this retrospective analysis, a total of 122 patients with locally advanced ESCC were included. These patients were randomly assigned to either the training cohort (n = 85) or the validation cohort (n = 37). In the training group, the least absolute shrinkage and selection operator (LASSO) regression was utilized to choose the best radiomic features for calculating the Rad-score. To develop a nomogram model, both univariate and multivariate analyses were conducted to identify the clinical factors and hematologic parameters that could predict the OS. The performance of the predictive model was evaluated using the C-index, while the accuracy was assessed through the calibration curve. RESULTS: The Rad-score was calculated by selecting 10 radiomic features through LASSO regression. OS was predicted independently by neutrophil-to-monocyte ratio (NMR) and Rad-score according to the results of multivariate analysis. Patients who had a Rad-score > 0.47 and an NMR > 9.76 were at a significant risk of mortality. A nomogram was constructed using the findings from the multivariate analysis. In the training cohort, the nomogram had a C-index of 0.619, while in the validation cohort, it was 0.573. The model's accuracy was demonstrated by the calibration curve, which was excellent. CONCLUSION: A prognostic model utilizing radiomics and hematologic parameters was developed, enabling the prediction of OS in patients with ESCC following dCRT. CRITICAL RELEVANCE STATEMENT: Patients with esophageal cancer who underwent definitive chemoradiotherapy may benefit from including CT radiomics in the nomogram model. KEY POINTS: ⢠Predicting the prognosis of ESCC patients before treatment is particularly important. ⢠Patients with a Rad-score > 0.47 and neutrophil-to-monocyte ratio > 9.76 had a high risk of mortality. ⢠CT-based radiomics nomogram model could be used to predict the survival of patients.
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Current semi-supervised video object segmentation (VOS) methods often employ the entire features of one frame to predict object masks and update memory. This introduces significant redundant computations. To reduce redundancy, we introduce a Region Aware Video Object Segmentation (RAVOS) approach, which predicts regions of interest (ROIs) for efficient object segmentation and memory storage. RAVOS includes a fast object motion tracker to predict object ROIs in the next frame. For efficient segmentation, object features are extracted based on the ROIs, and an object decoder is designed for object-level segmentation. For efficient memory storage, we propose motion path memory to filter out redundant context by memorizing the features within the motion path of objects. In addition to RAVOS, we also propose a large-scale occluded VOS dataset, dubbed OVOS, to benchmark the performance of VOS models under occlusions. Evaluation on DAVIS and YouTube-VOS benchmarks and our new OVOS dataset show that our method achieves state-of-the-art performance with significantly faster inference time, e.g., 86.1 J & F at 42 FPS on DAVIS and 84.4 J & F at 23 FPS on YouTube-VOS. Project page: ravos.netlify.app.
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Chloramphenicol (CAP) is an antibiotic that commonly pollutes the environment, and microorganisms primarily drive its degradation and transformation. Although several pathways for CAP degradation have been documented in different bacteria, multiple metabolic pathways in the same strain and their potential biological significance have not been revealed. In this study, Sphingobium WTD-1, which was isolated from activated sludge, can completely degrade 100 mg/L CAP within 60 h as the sole energy source. UPLC-HRMS and HPLC analyses showed that three different pathways, including acetylation, hydroxyl oxidation, and oxidation (C1-C2 bond cleavage), are responsible for the metabolism of CAP. Importantly, acetylation and C3 hydroxyl oxidation reduced the cytotoxicity of the substrate to strain WTD-1, and the C1-C2 bond fracture of CAP generated the metabolite p-nitrobenzoic acid (PNBA) to provide energy for its growth. This indicated that the synergistic action of three metabolic pathways caused WTD-1 to be adaptable and able to degrade high concentrations of CAP in the environment. This study deepens our understanding of the microbial degradation pathway of CAP and highlights the biological significance of the synergistic metabolism of antibiotic pollutants by multiple pathways in the same strain.
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Cloranfenicol , Sphingomonadaceae , Cloranfenicol/metabolismo , Biodegradación Ambiental , Antibacterianos/metabolismo , Redes y Vías Metabólicas , Sphingomonadaceae/metabolismoRESUMEN
Metabolic changes have been linked to the development of inflammatory bowel disease (IBD), which includes colitis. Allulose, an endogenous bioactive monosaccharide, is vital to the synthesis of numerous compounds and metabolic processes within living organisms. Nevertheless, the precise biochemical mechanism by which allulose inhibits colitis remains unknown. Allulose is an essential and intrinsic protector of the intestinal mucosal barrier, as it maintains the integrity of tight junctions in the intestines, according to the current research. It is also important to know that there is a link between the severity of inflammatory bowel disease (IBD) and colorectal cancer (CRC), chemically-induced colitis in rodents, and lower levels of allulose in the blood. Mice with colitis, either caused by dextran sodium sulphate (DSS) or naturally occurring colitis in IL-10-/- mice, had less damage to their intestinal mucosa after being given allulose. Giving allulose to a colitis model starts a chain of reactions because it stops cathepsin B from ejecting and helps lysosomes stick together. This system effectively stops the activity of myosin light chain kinase (MLCK) when intestinal epithelial damage happens. This stops the breakdown of tight junction integrity and the start of mitochondrial dysfunction. To summarise, the study's findings have presented data that supports the advantageous impact of allulose in reducing the advancement of colitis. Its ability to stop the disruption of the intestinal barrier enables this. Therefore, allulose has potential as a medicinal supplement for treating colitis.
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Colitis , Enteritis , Fructosa , Enfermedades Inflamatorias del Intestino , Enfermedades Mitocondriales , Humanos , Ratones , Animales , Catepsina B/metabolismo , Células CACO-2 , Enfermedades Inflamatorias del Intestino/metabolismo , Colitis/inducido químicamente , Colitis/tratamiento farmacológico , Colitis/metabolismo , Mucosa Intestinal , Uniones Estrechas , Enfermedades Mitocondriales/metabolismo , Sulfato de Dextran/farmacología , Ratones Endogámicos C57BL , Modelos Animales de EnfermedadRESUMEN
Stage pIIIA/N2 non-small cell lung cancer (NSCLC) is primarily treated by complete surgical resection combined with neoadjuvant/adjuvant therapies. However, up to 40% of patients experience tumor recurrence. Here, we studied 119 stage pIIIA/N2 NSCLC patients who received complete surgery plus adjuvant chemotherapy (CT) or chemoradiotherapy (CRT). The paired tumor and resection margin samples were analyzed using next-generation sequencing (NGS). Although all patients were classified as negative resection margins by histologic methods, NGS revealed that 47.1% of them had molecularly positive surgical margins. Patients who tested positive for NGS-detected residual tumors had significantly shorter disease-free survival (DFS) (P = 0.002). Additionally, metastatic lymph node ratio, erb-b2 receptor tyrosine kinase 2 (ERBB2) mutations, and SWI/SNF-related, matrix-associated, actin-dependent regulator of chromatin, subfamily a, member 4 (SMARCA4) mutations were also independently associated with DFS. We used these four features to construct a COX model that could effectively estimate recurrence risk and prognosis. Notably, mutational profiling through broad-panel NGS could more sensitively detect residual tumors than the conventional histologic methods. Adjuvant CT and adjuvant CRT exhibited no significant difference in eliminating locoregional recurrence risk for stage pIIIA/N2 NSCLC patients with molecularly positive surgical margins.
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Carcinoma de Pulmón de Células no Pequeñas , Neoplasias Pulmonares , Recurrencia Local de Neoplasia , Humanos , Carcinoma de Pulmón de Células no Pequeñas/patología , Carcinoma de Pulmón de Células no Pequeñas/genética , Carcinoma de Pulmón de Células no Pequeñas/cirugía , Carcinoma de Pulmón de Células no Pequeñas/terapia , Carcinoma de Pulmón de Células no Pequeñas/metabolismo , Femenino , Persona de Mediana Edad , Masculino , Neoplasias Pulmonares/patología , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/cirugía , Pronóstico , Anciano , Recurrencia Local de Neoplasia/genética , Recurrencia Local de Neoplasia/patología , Márgenes de Escisión , Supervivencia sin Enfermedad , Estadificación de Neoplasias , Mutación , Adulto , Secuenciación de Nucleótidos de Alto RendimientoRESUMEN
Learning discriminative representation with limited training samples is emerging as an important yet challenging visual categorization task. While prior work has shown that incorporating self-supervised learning can improve performance, we found that the direct use of canonical metric in a Lie group is theoretically incorrect. In this article, we prove that a valid optimization measurement should be a canonical metric on Lie algebra. Based on the theoretical finding, this article introduces a novel self-supervised Lie algebra network (SLA-Net) representation learning framework. Via minimizing canonical metric distance between target and predicted Lie algebra representation within a computationally convenient vector space, SLA-Net avoids computing nontrivial geodesic (locally length-minimizing curve) metric on a manifold (curved space). By simultaneously optimizing a single set of parameters shared by self-supervised learning and supervised classification, the proposed SLA-Net gains improved generalization capability. Comprehensive evaluation results on eight public datasets show the effectiveness of SLA-Net for visual categorization with limited samples.
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PURPOSE: The residual cancer burden index (RCB) was proposed as a response evaluation criterion in breast cancer patients treated with Neoadjuvant Chemotherapy (NAC). This study evaluated the relevance of RCB with replase-free survival (RFS). METHODS: The clinical data of 254 breast cancer patients who received NAC between 2016 and 2020 were retrospectively collected. The relationship between clinicopathologic factors and RFS was evaluated using Cox proportional hazards regression models. RFS estimates were determined by Kaplan-Meier(K-M) analysis and compared using the log-rank test. Multivariate logistic regression analysis was used to evaluate the risk factors associated with RCB. Receiver operating characteristic (ROC) curves showed the potential of the RCB and MP grading systems as biomarkers for RFS. RESULTS: At a median follow-up of 52 months, 59 patients(23.23%) developed relapse. Multivariate Cox regression showed that older age (P = 0.022), high Pathologic T stage after NAC (P = 0.023) and a high RCB score(P = 0.003) were risk factors for relapse. The outcomes of the multivariate logistic analysis indicated that RCB 0 (pathologic complete response [pCR]) was associated with HER2-positive patients (P = 0.002) and triple-negative breast cancer (TNBC) patients (P = 0.013). In addition, the RCB and MP scoring systems served as prognostic markers for patients who received NAC, and their area under curves (AUCs) were 0.691 and 0.342, respectively. CONCLUSION: These data suggest that RCB can be equally applied to predict RFS in Chinese patients with NAC. The application of RCB may help guide the selection of treatment strategies.
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Neoplasias de la Mama , Neoplasias de la Mama Triple Negativas , Humanos , Femenino , Neoplasias de la Mama/patología , Pronóstico , Terapia Neoadyuvante , Neoplasia Residual/patología , Estudios Retrospectivos , Recurrencia Local de Neoplasia/tratamiento farmacológico , Neoplasias de la Mama Triple Negativas/patología , Recurrencia , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéuticoRESUMEN
The choroid plexus (ChP) of the brain plays a central role in orchestrating the recruitment of peripheral leukocytes into the central nervous system (CNS) through the blood-cerebrospinal fluid (BCSF) barrier in pathological conditions, thus offering a unique niche to diagnose CNS disorders. We explored whether magnetic resonance imaging of the ChP could be optimized for mild traumatic brain injury (mTBI). mTBI induces subtle, yet influential, changes in the brain and is currently severely underdiagnosed. We hypothesized that mTBI induces sufficient alterations in the ChP to cause infiltration of circulating leukocytes through the BCSF barrier and developed macrophage-adhering gadolinium [Gd(III)]-loaded anisotropic micropatches (GLAMs), specifically designed to image infiltrating immune cells. GLAMs are hydrogel-based discoidal microparticles that adhere to macrophages without phagocytosis. We present a fabrication process to prepare GLAMs at scale and demonstrate their loading with Gd(III) at high relaxivities, a key indicator of their effectiveness in enhancing image contrast and clarity in medical imaging. In vitro experiments with primary murine and porcine macrophages demonstrated that GLAMs adhere to macrophages also under shear stress and did not affect macrophage viability or functions. Studies in a porcine mTBI model confirmed that intravenously administered macrophage-adhering GLAMs provide a differential signal in the ChP and lateral ventricles at Gd(III) doses 500- to 1000-fold lower than those used in the current clinical standard Gadavist. Under the same mTBI conditions, Gadavist did not offer a differential signal at clinically used doses. Our results suggest that macrophage-adhering GLAMs could facilitate mTBI diagnosis.
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Conmoción Encefálica , Lesiones Traumáticas del Encéfalo , Animales , Ratones , Porcinos , Gadolinio , Lesiones Traumáticas del Encéfalo/diagnóstico por imagen , Encéfalo/patología , Imagen por Resonancia Magnética/métodos , Conmoción Encefálica/patología , Macrófagos/patologíaRESUMEN
The covalent attachment of polyethylene glycol (PEG) to therapeutic agents, termed PEGylation, is a well-established and clinically proven drug delivery approach to improve the pharmacokinetics and pharmacodynamics of drugs. Specifically, PEGylation can improve the parent drug's solubility, extend its circulation time, and reduce its immunogenicity, with minimal undesirable properties. PEGylation technology has been applied to various therapeutic modalities including small molecules, aptamers, peptides, and proteins, leading to over 30 PEGylated drugs currently used in the clinic and many investigational PEGylated agents under clinical trials. Here, we summarize the diverse types of PEGylation strategies, the key advantages of PEGylated therapeutics over their parent drugs, and the broad applications and impacts of PEGylation in clinical settings. A particular focus has been given to the size, topology, and functionalities of PEG molecules utilized in clinically used PEGylated drugs, as well as those under clinical trials. An additional section has been dedicated to analyzing some representative PEGylated drugs that were discontinued at different stages of clinical studies. Finally, we critically discuss the current challenges faced in the development and clinical translation of PEGylated agents.
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SNOMED CT is a comprehensive medical ontology used in health care sectors across the world covering a wide range of concepts that support diversity at the point of healthcare. However, not all these concepts are needed for every use case; it is better to concentrate on those parts that apply to the particular application while preserving the meaning of relevant concepts. This paper considers the application of a novel subontology extraction method to create a new resource, called the IPS terminology, which functions as a standalone ontology with the same features as SNOMED CT, but is designed for cross-border patient care. The IPS terminology has been released for free use under an open license, with the intention of promoting interoperability of health information worldwide.
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Sector de Atención de Salud , Instituciones de Salud , Humanos , Intención , Systematized Nomenclature of MedicineAsunto(s)
Neoplasias Pulmonares , Carcinoma Pulmonar de Células Pequeñas , Humanos , Carcinoma Pulmonar de Células Pequeñas/tratamiento farmacológico , Carcinoma Pulmonar de Células Pequeñas/genética , Neoplasias Pulmonares/tratamiento farmacológico , Neoplasias Pulmonares/genética , Multiómica , Radiómica , GenómicaRESUMEN
Automatic detection of retinal vasculature in optical coherence tomography angiography (OCTA) images faces several challenges such as the closely located capillaries, vessel discontinuity and high noise level. This paper introduces a new distinctive phase interdependency model to address these problems for delineating centerline patterns of the vascular network. We capture the inherent property of vascular centerlines by obtaining the inter-scale dependency information that exists between neighboring symmetrical wavelets in complex Poisson domain. In particular, the proposed phase interdependency model identifies vascular centerlines as the distinctive features that have high magnitudes over adjacent symmetrical coefficients whereas the coefficients caused by background noises are decayed rapidly along adjacent wavelet scales. The potential relationships between the neighboring Poisson coefficients are established based on the coherency of distinctive symmetrical wavelets. The proposed phase model is assessed on the OCTA-500 database (300 OCTA images + 200 OCT images), ROSE-1-SVC dataset (9 OCTA images), ROSE-1 (SVC+ DVC) dataset (9 OCTA images), and ROSE-2 dataset (22 OCTA images). The experiments on the clinically relevant OCTA images validate the effectiveness of the proposed method in achieving high-quality results. Our method produces average FScore of 0.822, 0.782, and 0.779 on ROSE-1-SVC, ROSE-1 (SVC+ DVC), and ROSE-2 datasets, respectively, and the FScore of 0.910 and 0.862 on OCTA_6mm and OCT_3mm datasets (OCTA-500 database), respectively, demonstrating its superior performance over the state-of-the-art benchmark methods.
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Angiografía , Vasos Retinianos , Vasos Retinianos/diagnóstico por imagen , Capilares , Tomografía de Coherencia Óptica/métodos , Angiografía con Fluoresceína/métodosRESUMEN
Chloramphenicols (CAPs) are ubiquitous emerging pollutants that threaten ecological environments and human health. Microbial and enzyme-based biodegradation strategies offer a cost-effective environmentally friendly approach for CAPs removal from contaminated sites. Here, CpmO, a novel multifunctional oxidase for CAP degradation was identified from the CAP-degrading strain Sphingobium sp. WTD-1. This enzyme was found to be responsible for both the oxidation of the C3-hydroxyl and oxidative cleavage of the C1-C2 bond of CAP, and the oxidative cleavage pathway of CAP was dominant. The catalytic efficiency of CpmO for CAP was 41.6 times that for thiamphenicol (TAP) under the optimal conditions (40 °C, pH 6.0). CpmO was identified as a member of the glucose-methanol-choline oxidoreductase family. Molecular docking and site-directed mutagenesis analysis indicated that CAP was connected to the key amino acid residues E231/E395, K277, and I273/A276 in CpmO through hydrogen bonding, nonclassical hydrogen bonding, and π-π stacking forces, respectively. The catalytic activities of the A276W, K277P, and E231S mutants were found to be 1.1 times, 6.4 times, and 13.2 times higher than that of the wild type, respectively. These findings provide genetic resources and theoretical guidance for future application in biotechnological and metabolic engineering efforts for the remediation of CAPs-contaminated environments.
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Cloranfenicol , Oxidorreductasas , Humanos , Sitios de Unión/genética , Simulación del Acoplamiento Molecular , Mutagénesis Sitio-DirigidaRESUMEN
Elevated evidence has reported the important role of oxidative stress injury and inflammatory response in the progression of colitis. Tumor Suppressor TSBF1, TRIM59, is a ubiquitin E3 ligase and mediates immune response. However, the underlying molecular function of TRIM59 on regulation of colitis is still not understood. In the current study, we identify the TRIM59 as a critical and novel endogenous suppressor of kelch-like ECH-associated protein 1 (KEAP1), and we also determine that TRIM59 is a KEAP1-interacting partner protein that catalyses its ubiquitination and degradation in intestinal epithelial cells (IEC). Moreover, IEC-specific loss of the Trim59 disrupts colon metabolic homeostasis, accompanied by intestinal oxidative stress injury, elevated endogenous reactive oxygen species (ROS) production and pro-inflammatory cytokines release, significantly promotes acute or chronic colitis progression. Conversely, transgenic mice with Trim59 overexpression by adeno-associated virus (AAV)-induced Trim59 gene therapeutics mitigates colitis in acute or chronic colitis rodent models and in vitro experiments. Mechanistically, in response to onset of colitis, TRIM59 directly interacts with KEAP1 and promotes ubiquitin-proteasome degradation, thus results in NRF2 activation and its downstream cascade anti-oxidative stress-related pathway activation, which facilitates anti-oxidant defense and reduces tissue damage. All the findings elucidated the potential role of TRIM59 in colitis progression by mediating KEAP1 deactivation and degradation, and could be considered as a therapeutic target for the treatment of such disease.
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Colitis , Factor 2 Relacionado con NF-E2 , Animales , Ratones , Enfermedad Crónica , Proteína 1 Asociada A ECH Tipo Kelch/genética , Proteína 1 Asociada A ECH Tipo Kelch/metabolismo , Ratones Transgénicos , Factor 2 Relacionado con NF-E2/genética , Factor 2 Relacionado con NF-E2/metabolismo , Estrés Oxidativo , Transducción de Señal , Ubiquitina/metabolismo , Ubiquitina/farmacologíaRESUMEN
Uncontrolled bleeding is a life-threatening emergency that requires immediate intervention. Currently available on-site bleeding interventions largely rely on the use of tourniquets, pressure dressing, and other topical hemostatic agents, which can only treat bleeding injuries that are known, accessible, and potentially compressible. Synthetic hemostats that are stable at room temperature, easy to carry, field-usable, and able to stop internal bleeding at multiple or unknown sources, are still lacking. We recently developed a hemostatic agent via polymer peptide interfusion (HAPPI), which can selectively bind to activated platelets and injury sites after intravascular administration. Here we report that HAPPI is highly effective in treating multiple lethal traumatic bleeding conditions in normal as well as hemophilia models via either systemic administration or topical application. In a rat liver traumatic model, intravenous injection of HAPPI resulted in a significant decrease in blood loss and a four-fold reduction in mortality rate within 2 h after injury. When applied topically on liver punch biopsy wounds in heparinized rats, HAPPI achieved a 73% of reduction in blood loss and a five-fold increase in survival rate. HAPPI also exhibited hemostatic efficacy in hemophilia A mice by reducing blood loss. Further, HAPPI worked synergistically with rFVIIa to induce immediate hemostasis and 95% reduction in total blood loss compared to the saline-treated group in hemophelia mice models. These results demonstrate that HAPPI is a promising field-usable hemostatic agent for a broad range of different hemorrhagic conditions.