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1.
Nat Chem Biol ; 2024 May 21.
Artigo em Inglês | MEDLINE | ID: mdl-38773328

RESUMO

A timely inflammatory response is crucial for early viral defense, but uncontrolled inflammation harms the host. Retinoic acid-inducible gene I (RIG-I) has a pivotal role in detecting RNA viruses, yet the regulatory mechanisms governing its sensitivity remain elusive. Here we identify PTENα, an N-terminally extended form of PTEN, as an RNA-binding protein with a preference for the CAUC(G/U)UCAU motif. Using both in vivo and in vitro viral infection assays, we demonstrated that PTENα restricted the host innate immune response, relying on its RNA-binding capacity and phosphatase activity. Mechanistically, PTENα directly bound to viral RNA and enzymatically converted its 5'-triphosphate to 5'-monophosphate, thereby reducing RIG-I sensitivity. Physiologically, brain-intrinsic PTENα exerted protective effects against viral inflammation, while peripheral PTENα restricted host antiviral immunity and, to some extent, promoted viral replication. Collectively, our findings underscore the significance of PTENα in modulating viral RNA- and RIG-I-mediated immune recognition, offering potential therapeutic implications for infectious diseases.

2.
Int J Biol Macromol ; 270(Pt 1): 132432, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38761609

RESUMO

The African swine fever virus (ASFV) continues to pose significant economic and pandemic risks. Consequently, discovering new, efficient vaccines is crucial. Messenger RNA (mRNA) vaccines have emerged as promising candidates, providing minimal risk of insertional mutagenesis, high safety profiles, effectiveness, rapid scalability in production, and cost-effectiveness. In this study, we have developed an ASF p30 mRNA vaccine candidate (mRNA/Man-LNP) employing mannose-modified lipid nanoparticles (LNPs). The mRNA/Man-LNP exhibited effective antigen presentation and facilitated dendritic cells (DCs) maturation. Notably, it elicited strong IgG titers and activated CD4+ and CD8+ T-cells in immunized mice, all while adhering to stringent biosafety standards. This investigation demonstrates that mRNA/Man-LNP can trigger both humoral and cellular immune responses, suggesting its potential as a potent and promising vaccine candidate for controlling African swine fever (ASF).


Assuntos
Vírus da Febre Suína Africana , Febre Suína Africana , Manose , Nanopartículas , Vacinas Virais , Animais , Nanopartículas/química , Vírus da Febre Suína Africana/imunologia , Vírus da Febre Suína Africana/genética , Febre Suína Africana/prevenção & controle , Febre Suína Africana/imunologia , Camundongos , Vacinas Virais/imunologia , Suínos , Manose/química , Células Dendríticas/imunologia , Lipídeos/química , Desenvolvimento de Vacinas , RNA Mensageiro/genética , RNA Mensageiro/imunologia , Vacinas de mRNA , Feminino , Anticorpos Antivirais/imunologia , Anticorpos Antivirais/sangue , Lipossomos
3.
Metabolomics ; 20(3): 59, 2024 May 21.
Artigo em Inglês | MEDLINE | ID: mdl-38773019

RESUMO

INTRODUCTION: Thyroid cancer incidence rate has increased substantially worldwide in recent years. Fine needle aspiration biopsy (FNAB) is currently the golden standard of thyroid cancer diagnosis, which however, is invasive and costly. In contrast, breath analysis is a non-invasive, safe and simple sampling method combined with a promising metabolomics approach, which is suitable for early cancer diagnosis in high volume population. OBJECTIVES: This study aims to achieve a more comprehensive and definitive exhaled breath metabolism profile in papillary thyroid cancer patients (PTCs). METHODS: We studied both end-tidal and mixed expiratory breath, solid-phase microextraction gas chromatography coupled with high resolution mass spectrometry (SPME-GC-HRMS) was used to analyze the breath samples. Multivariate combined univariate analysis was applied to identify potential breath biomarkers. RESULTS: The biomarkers identified in end-tidal and mixed expiratory breath mainly included alkanes, olefins, enols, enones, esters, aromatic compounds, and fluorine and chlorine containing organic compounds. The area under the curve (AUC) values of combined biomarkers were 0.974 (sensitivity: 96.1%, specificity: 90.2%) and 0.909 (sensitivity: 98.0%, specificity: 74.5%), respectively, for the end-tidal and mixed expiratory breath, indicating of reliability of the sampling and analysis method CONCLUSION: This work not only successfully established a standard metabolomic approach for early diagnosis of PTC, but also revealed the necessity of using both the two breath types for comprehensive analysis of the biomarkers.


Assuntos
Biomarcadores Tumorais , Testes Respiratórios , Cromatografia Gasosa-Espectrometria de Massas , Metabolômica , Microextração em Fase Sólida , Câncer Papilífero da Tireoide , Neoplasias da Glândula Tireoide , Humanos , Metabolômica/métodos , Câncer Papilífero da Tireoide/diagnóstico , Câncer Papilífero da Tireoide/metabolismo , Testes Respiratórios/métodos , Cromatografia Gasosa-Espectrometria de Massas/métodos , Microextração em Fase Sólida/métodos , Feminino , Masculino , Pessoa de Meia-Idade , Biomarcadores Tumorais/análise , Biomarcadores Tumorais/metabolismo , Adulto , Neoplasias da Glândula Tireoide/diagnóstico , Neoplasias da Glândula Tireoide/metabolismo , Detecção Precoce de Câncer/métodos , Idoso
4.
Nat Commun ; 15(1): 2906, 2024 Apr 04.
Artigo em Inglês | MEDLINE | ID: mdl-38575578

RESUMO

Mechano-sensitive hair-like sensilla (MSHS) have an ingenious and compact three-dimensional structure and have evolved widely in living organisms to perceive multidirectional mechanical signals. Nearly all MSHS are iontronic or electronic, including their biomimetic counterparts. Here, an all-optical mechano-sensor mimicking MSHS is prototyped and integrated based on a thin-walled glass microbubble as a flexible whispering-gallery-mode resonator. The minimalist integrated device has a good directionality of 32.31 dB in the radial plane of the micro-hair and can detect multidirectional displacements and forces as small as 70 nm and 0.9 µN, respectively. The device can also detect displacements and forces in the axial direction of the micro-hair as small as 2.29 nm and 3.65 µN, respectively, and perceive different vibrations. This mechano-sensor works well as a real-time, directional mechano-sensory whisker in a quadruped cat-type robot, showing its potential for innovative mechano-transduction, artificial perception, and robotics applications.


Assuntos
Robótica , Sensilas , Animais , Cabelo , Fenômenos Mecânicos , Eletrônica
5.
Dalton Trans ; 53(16): 6965-6973, 2024 Apr 23.
Artigo em Inglês | MEDLINE | ID: mdl-38546786

RESUMO

Many diseases are due to bacterial infections, which are treated by penicillin. Existing methods for penicillin detection have relatively high requirements for sample storage and processing, personnel professionalism, and instruments. Herein, water-soluble N-C quantum dots (QDs) from wheat straw were synthesized in a green way by using an efficient and simple method. The N-C QDs were modified with an imprinted layer by a gel-sol method. Penicillin selectively quenched the fluorescence emission of N-C QDs@MIP, and a linear relationship was obtained in the concentration range of 1.0 × 10-6-15.2 × 10-6 mol L-1. The reliability of the sensor in real sample analysis was satisfactory with results in the range of 93.6%-100%, and the sensor showed good reproducibility and long-term stability. The study provides a simple strategy to fabricate N-C QDs@MIP with a highly selective recognition ability and opens an avenue to develop highly efficient sensing probes for the detection of antibiotics in biological applications.

6.
J Nucl Med ; 65(5): 728-734, 2024 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-38514084

RESUMO

Immune checkpoint blockade (ICB) has achieved groundbreaking results in clinical cancer therapy; however, only a subset of patients experience durable benefits. The aim of this study was to explore strategies for predicting tumor responses to optimize the intervention approach using ICB therapy. Methods: We used a bilateral mouse model for proteomics analysis to identify new imaging biomarkers for tumor responses to ICB therapy. A PET radiotracer was synthesized by radiolabeling the identified biomarker-targeting antibody with 124I. The radiotracer was then tested for PET prediction of tumor responses to ICB therapy. Results: We identified galectin-1 (Gal-1), a member of the carbohydrate-binding lectin family, as a potential negative biomarker for ICB efficacy. We established that Gal-1 inhibition promotes a sensitive immune phenotype within the tumor microenvironment (TME) for ICB therapy. To assess the pre-ICB treatment status of the TME, a Gal-1-targeted PET radiotracer, 124I-αGal-1, was developed. PET imaging with 124I-αGal-1 showed the pretreatment immunosuppressive status of the TME before the initiation of therapy, thus enabling the prediction of ICB resistance in advance. Moreover, the use of hydrogel scaffolds loaded with a Gal-1 inhibitor, thiodigalactoside, demonstrated that a single dose of thiodigalactoside-hydrogel significantly potentiated ICB and adoptive cell transfer immunotherapies by remodeling the immunosuppressive TME. Conclusion: Our study underscores the potential of Gal-1-targeted PET imaging as a valuable strategy for early-stage monitoring of tumor responses to ICB therapy. Additionally, Gal-1 inhibition effectively counteracts the immunosuppressive TME, resulting in enhanced immunotherapy efficacy.


Assuntos
Galectina 1 , Imunoterapia , Tomografia por Emissão de Pósitrons , Microambiente Tumoral , Galectina 1/metabolismo , Animais , Camundongos , Linhagem Celular Tumoral , Inibidores de Checkpoint Imunológico/uso terapêutico , Inibidores de Checkpoint Imunológico/farmacologia , Feminino , Resultado do Tratamento , Radioisótopos do Iodo , Humanos
7.
EMBO J ; 43(6): 1089-1109, 2024 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-38360992

RESUMO

Cullin-RING E3 ubiquitin ligase (CRL) family members play critical roles in numerous biological processes and diseases including cancer and Alzheimer's disease. Oligomerization of CRLs has been reported to be crucial for the regulation of their activities. However, the structural basis for its regulation and mechanism of its oligomerization are not fully known. Here, we present cryo-EM structures of oligomeric CRL2FEM1B in its unneddylated state, neddylated state in complex with BEX2 as well as neddylated state in complex with FNIP1/FLCN. These structures reveal that asymmetric dimerization of N8-CRL2FEM1B is critical for the ubiquitylation of BEX2 while FNIP1/FLCN is ubiquitylated by monomeric CRL2FEM1B. Our data present an example of the asymmetric homo-dimerization of CRL. Taken together, this study sheds light on the ubiquitylation strategy of oligomeric CRL2FEM1B according to substrates with different scales.


Assuntos
Ubiquitina-Proteína Ligases , Humanos , Proteínas Culina/metabolismo , Neoplasias/metabolismo , Proteínas do Tecido Nervoso , Ubiquitina/metabolismo , Ubiquitina-Proteína Ligases/metabolismo , Ubiquitinação
8.
ACS Nano ; 18(8): 6477-6486, 2024 Feb 27.
Artigo em Inglês | MEDLINE | ID: mdl-38350867

RESUMO

Enhancing light-matter interaction is a key requisite in the realm of optical sensors. Bound states in the continuum (BICs), possessing high quality factors (Q factors), have shown great advantages in sensing applications. Recent theories elucidate the ability of BICs with hybrid metal-dielectric architectures to achieve high Q factors and high sensitivities. However, the experimental validation of the sensing performance in such hybrid systems remains equivocal. In this study, we propose two symmetry-protected quasi-BIC modes in a metal-dielectric metasurface. Our results demonstrate that, under the normal incidence of light, the quasi-BIC mode dominated by dielectric can achieve a high Q factor of 412 and a sensing performance with a high bulk sensitivity of 492.7 nm/RIU (refractive index unit) and a figure of merit (FOM) of 266.3 RIU-1, while the quasi-BIC mode dominated by metal exhibits a stronger surface affinity in the biotin-streptavidin bioassay. These findings offer a promising approach for implementing metasurface-based sensors, representing a paradigm for high-sensitivity biosensing platforms.

9.
J Genet Genomics ; 51(5): 531-542, 2024 May.
Artigo em Inglês | MEDLINE | ID: mdl-38184105

RESUMO

Phosphatase and tensin homolog (PTEN) is a multifunctional gene involved in a variety of physiological and pathological processes. Circular RNAs (circRNAs) are generated from back-splicing events during mRNA processing and participate in cell biological processes through binding to RNAs or proteins. However, PTEN-related circRNAs are largely unknown. Here, we report that circPTEN- mitochondria (MT) (hsa_circ_0002934) is a circular RNA encoded by exons 3, 4, and 5 of PTEN and is a critical regulator of mitochondrial energy metabolism. CircPTEN-MT is localized to mitochondria and physically associated with leucine-rich pentatricopeptide repeat-containing protein (LRPPRC), which regulates posttranscriptional gene expression in mitochondria. Knocking down circPTEN-MT reduces the interaction of LRPPRC and steroid receptor RNA activator (SRA) stem-loop interacting RNA binding protein (SLIRP) and inhibits the polyadenylation of mitochondrial mRNA, which decreases the mRNA level of the mitochondrial complex I subunit and reduces mitochondrial membrane potential and adenosine triphosphate production. Our data demonstrate that circPTEN-MT is an important regulator of cellular energy metabolism. This study expands our understanding of the role of PTEN, which produces both linear and circular RNAs with different and independent functions.


Assuntos
Metabolismo Energético , Mitocôndrias , PTEN Fosfo-Hidrolase , RNA Circular , Proteínas de Ligação a RNA , PTEN Fosfo-Hidrolase/genética , PTEN Fosfo-Hidrolase/metabolismo , RNA Circular/genética , RNA Circular/metabolismo , Humanos , Mitocôndrias/genética , Mitocôndrias/metabolismo , Metabolismo Energético/genética , Proteínas de Ligação a RNA/genética , Proteínas de Ligação a RNA/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Potencial da Membrana Mitocondrial/genética , Complexo I de Transporte de Elétrons/genética , Complexo I de Transporte de Elétrons/metabolismo , Trifosfato de Adenosina/metabolismo , Células HEK293 , Proteínas de Neoplasias
10.
J Exp Bot ; 75(1): 258-273, 2024 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-37721809

RESUMO

Intercropping improves resource utilization. Under wide-narrow-row maize (Zea mays) intercropping, maize plants are subjected to weak unilateral illumination and exhibit high photosynthetic performance. However, the mechanism regulating photosynthesis under unilateral weak light remains unknown. We investigated the relationship between photosynthesis and sugar metabolism in maize under unilateral weak light. Our results showed that the net photosynthetic rate (Pn) of unshaded leaves increased as the level of shade on the other side increased. On the contrary, the concentration of sucrose and starch and the number of starch granules in the unshaded leaves decreased with increased shading due to the transfer of abundant C into the grains. However, sink loss with ear removal reduced the Pn of unshaded leaves. Intense unilateral shade (40% to 20% normal light), but not mild unilateral shade (60% normal light), reduced grain yield (37.6% to 54.4%, respectively). We further found that in unshaded leaves, Agpsl, Bmy, and Mexl-like expression significantly influenced sucrose and starch metabolism, while Sweet13a and Sut1 expression was crucial for sugar export. In shaded leaves, expression of Sps1, Agpsl, and Sweet13c was crucial for sugar metabolism and export. This study confirmed that unshaded leaves transported photosynthates to the ear, leading to a decrease in sugar concentration. The improvement of photosynthetic performance was associated with altered sugar transport. We propose a narrow-row spacing of 40 cm, which provides appropriate unilateral shade and limits yield reduction.


Assuntos
Fotossíntese , Zea mays , Fotossíntese/fisiologia , Zea mays/fisiologia , Folhas de Planta/fisiologia , Amido , Sacarose
11.
Artigo em Inglês | MEDLINE | ID: mdl-38082811

RESUMO

For focal epilepsy patients, correctly identifying the seizure onset zone (SOZ) is essential for surgical treatment. In automated realistic SOZ identification, it is necessary to identify the SOZ of an unknown patient using another patient's electroencephalogram (EEG). However, in such cases, the influence of individual differences in EEG becomes a bottleneck. In this paper, we propose the method with domain adaptation and source patient selection to address the issue of individual differences in EEG and improve performance. The proposed method was evaluated on intracranial EEG data from 11 patients with epilepsy caused by focal cortical dysplasia. The results showed that the proposed method significantly improved SOZ identification performance compared to existing methods without domain adaptation and source patient selection. In addition, it was suggested that data from residual-seizure patients may have adversely affected estimation performance. Visualization of the prediction on MRI images showed that the proposed method might detect SOZs missed by epileptologists.


Assuntos
Encéfalo , Epilepsias Parciais , Humanos , Eletrocorticografia , Eletroencefalografia/métodos , Convulsões/diagnóstico
12.
Cell Rep ; 42(11): 113388, 2023 11 28.
Artigo em Inglês | MEDLINE | ID: mdl-37934668

RESUMO

Cancer evades host immune surveillance by virtue of poor immunogenicity. Here, we report an immune suppressor, designated as PTIR1, that acts as a promotor of tumor immune resistance. PTIR1 is selectively induced in human cancers via alternative splicing of DDX58 (RIG-I), and its induction is closely related to poor outcome in patients with cancer. Through blocking the recruitment of leukocytes, PTIR1 facilitates cancer immune escape and tumor-intrinsic resistance to immunotherapeutic treatments. Unlike RIG-I, PTIR1 is capable of binding to the C terminus of UCHL5 and activates its ubiquitinating function, which in turn inhibits immunoproteasome activity and limits neoantigen processing and presentation, consequently blocking T cell recognition and attack against cancer. Moreover, we find that the adenosine deaminase ADAR1 induces A-to-I RNA editing on DDX58 transcript, thus triggering PTIR1 production. Collectively, our data uncover the immunosuppressive role of PTIR1 in tumorigenesis and propose that ADAR1-PTIR1-UCHL5 signaling is a potential cancer immunotherapeutic target.


Assuntos
Comunicação Celular , Transdução de Sinais , Humanos , Carcinogênese/genética , Isoformas de Proteínas/genética , Isoformas de Proteínas/metabolismo , Adenosina Desaminase/genética , Adenosina Desaminase/metabolismo , Proteína DEAD-box 58/metabolismo , Receptores Imunológicos , Ubiquitina Tiolesterase/genética , Ubiquitina Tiolesterase/metabolismo
13.
J Virol ; 97(12): e0157423, 2023 Dec 21.
Artigo em Inglês | MEDLINE | ID: mdl-38014947

RESUMO

IMPORTANCE: Marek's disease virus (MDV) is a highly infectious and oncogenic virus that can induce severe T cell lymphomas in chickens. MDV encodes more than 100 genes, most of which have unknown functions. This work indicated that the LORF9 gene is necessary for MDV early cytolytic replication in B lymphocytes. In addition, we have found that the LORF9 deletion mutant has a comparative immunological protective effect with CVI988/Rispens vaccine strain against very virulent MDV challenge. This is a significant discovery that LORF9 can be exploited as a possible target for the development of an MDV gene deletion vaccine.


Assuntos
Herpesvirus Galináceo 2 , Vacinas contra Doença de Marek , Doença de Marek , Doenças das Aves Domésticas , Animais , Linfócitos B , Galinhas , Deleção de Genes , Herpesvirus Galináceo 2/genética , Doença de Marek/prevenção & controle , Doença de Marek/genética , Vacinas contra Doença de Marek/genética , Replicação Viral
14.
Heliyon ; 9(11): e21847, 2023 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-38034812

RESUMO

Lead (Pb) is a heavy metal that can have harmful effects on the environment, which has severe cytotoxicity in many animal tissues. N-acetylcysteine (NAC) has antioxidant activity, reducing lead-induced oxidative stress and apoptosis, but its role in chicken cells is unknown. The current study explored the antagonistic effect of NAC on lead-induced apoptosis and oxidative stress in chicken embryo fibroblast (CEF). In this study, CEF was used as a model to measure the cytotoxic effects of lead nitrate at different concentrations, demonstrating a dose-dependent effect on CEF activity. Employing inverted microscopy, the investigation of morphological alterations in CEF cells was conducted. Fluorescence staining methodology enabled the assessment of reactive oxygen species (ROS) levels within CEF cells. Moreover, an enzyme-linked immunosorbent assay was utilized to detect the presence of oxidative damage indicators encompassing superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), catalase (CAT) activity, malondialdehyde (MDA) content, and total antioxidant capacity (T-AOC) within CEF cells. Furthermore, the determination of the apoptosis rate of CEF cells was accomplished through the utilization of the Hoechst 33258 staining method in combination with the Annexin V-FITC dual staining method. By using RT-qPCR for detection, lead treatment increased expression of pro-apoptotic genes, caspase-3, and caspase-9, and reduced expression of anti-apoptotic genes, Bcl-2, and BI-1. Reduced antioxidant capacity was shown by increased ROS and MDA levels in CEF cells after lead treatment. The results showed that NAC inhibited the expression of caspase-3 and caspase-9 in lead-treated CEF cells, while NAC had a certain inhibitory effect on the relative expression of Bcl-2 and BI-1 mRNA in lead-induced CEF cells. NAC significantly reduced lead-induced oxidative damage and apoptosis. Overall, our results demonstrate a novel protective effect of NAC against lead-induced injury in chicken cells, providing a theoretical basis for future investigations of drugs that are effective in preventing lead poisoning in animals.

15.
Biosensors (Basel) ; 13(10)2023 Sep 28.
Artigo em Inglês | MEDLINE | ID: mdl-37887104

RESUMO

A novel multi-optofluidic microbubble resonator (OMBR) sensitivity equalization method is presented that equalizes the sensing signal from different OMBRs. The method relies on the fact that the ratio of the wavelength shifts to the bulk refractive index sensitivity (BRIS) does not depend on the physical dimensions of the OMBR. The proof of concept is experimentally validated and the sensing signals from individual OMBRs can be directly compared. Furthermore, a wide dynamic range of sensing with favorable consistency and repeatability is achieved by piecing together signals from 20 OMBRs for HIV-1 p24 antigen detection from 50 fg/mL to 100 ng/mL (2.1 fM to 4.2 nM), indicating significant potential for practical applications, such as in drug screening and disease diagnosis.


Assuntos
Técnicas Biossensoriais , Microbolhas , Técnicas Biossensoriais/métodos , Refratometria
16.
Cell Death Dis ; 14(10): 680, 2023 10 13.
Artigo em Inglês | MEDLINE | ID: mdl-37833251

RESUMO

Nephrolithiasis is highly prevalent and associated with the increased risk of kidney cancer. The tumor suppressor von Hippel-Lindau (VHL) is critical for renal cancer development, however, its role in kidney stone disease has not been fully elucidated until now. Here we reported VHL expression was upregulated in renal epithelial cells upon exposure to crystal. Utilizing Vhl+/mu mouse model, depletion of VHL exacerbated kidney inflammatory injury during nephrolithiasis. Conversely, overexpression of VHL limited crystal-induced lipid peroxidation and ferroptosis in a BICD2-depdendent manner. Mechanistically, VHL interacted with the cargo adaptor BICD2 and promoted itsd K48-linked poly-ubiquitination, consequently resulting in the proteasomal degradation of BICD2. Through promoting STAT1 nuclear translocation, BICD2 facilitated IFNγ signaling transduction and enhanced IFNγ-mediated suppression of cystine/glutamate antiporter system Xc-, eventually increasing cell sensitivity to ferroptosis. Moreover, we found that the BRAF inhibitor impaired the association of VHL with BICD2 through triggering BICD2 phosphorylation, ultimately causing severe ferroptosis and nephrotoxicity. Collectively, our results uncover the important role of VHL/BICD2/STAT1 axis in crystal kidney injury and provide a potential therapeutic target for treatment and prevention of renal inflammation and drug-induced nephrotoxicity.


Assuntos
Carcinoma de Células Renais , Neoplasias Renais , Nefrolitíase , Animais , Camundongos , Proteína Supressora de Tumor Von Hippel-Lindau/genética , Proteína Supressora de Tumor Von Hippel-Lindau/metabolismo , Rim/patologia , Neoplasias Renais/metabolismo , Carcinoma de Células Renais/metabolismo , Nefrolitíase/metabolismo
17.
Int J Biol Macromol ; 253(Pt 7): 127311, 2023 Dec 31.
Artigo em Inglês | MEDLINE | ID: mdl-37865977

RESUMO

African Swine Fever (ASF) is an acute and highly lethal disease in pigs caused by African Swine Fever Virus (ASFV). Viral proteins have been commonly used as antigenic targets for the development of ASF diagnostic methods. However, the prokaryotic expression of viral proteins has deficiencies such as instability, insolubility, and high cost in eukaryotic situations. This study screened and verified ASFV-encoded p72, p54, and p30 protein antigenic epitopes. Subsequently, a novel antigenic epitope-associated recombinant protein was designed based on an ideal structural protein and expressed in Escherichia coli (E. coli). Western blot analysis indicated that the recombinant protein could specifically react with the monoclonal antibody (mAb) of p72 and polyclonal antibodies of p54 and p30, respectively. Next, an ASF indirect ELISA (iELISA) method was established based on the recombinant protein, which has no specific reaction with sera of other important pig viral diseases. Meanwhile, it shows a sensitivity to detecting dilutions of ASF-positive reference serum up to 1:6400. The clinical sample detection results showed a high coincidence rate of 98 % with a commercial competition ELISA kit. In conclusion, we established a novel specific, and sensitive ASF serologic detection method that opens new avenues for ASF serodiagnostic method development.


Assuntos
Vírus da Febre Suína Africana , Febre Suína Africana , Suínos , Animais , Febre Suína Africana/diagnóstico , Epitopos , Escherichia coli/genética , Proteínas Recombinantes/genética , Proteínas Virais , Ensaio de Imunoadsorção Enzimática/métodos , Anticorpos Antivirais
18.
Neuron ; 111(22): 3634-3649.e7, 2023 11 15.
Artigo em Inglês | MEDLINE | ID: mdl-37683629

RESUMO

Blood-brain barrier (BBB) function deteriorates during aging, contributing to cognitive impairment and neurodegeneration. It is unclear what drives BBB leakage in aging and how it can be prevented. Using single-nucleus transcriptomics, we identified decreased connexin 43 (CX43) expression in cadherin-5+ (Cdh5+) cerebral vascular cells in naturally aging mice and confirmed it in human brain samples. Global or Cdh5+ cell-specific CX43 deletion in mice exacerbated BBB dysfunction during aging. The CX43-dependent effect was not due to its canonical gap junction function but was associated with reduced NAD+ levels and mitochondrial dysfunction through NAD+-dependent sirtuin 3 (SIRT3). CX43 interacts with and negatively regulates poly(ADP-ribose) polymerase 1 (PARP1). Pharmacologic inhibition of PARP1 by olaparib or nicotinamide mononucleotide (NMN) supplementation rescued NAD+ levels and alleviated aging-associated BBB leakage. These findings establish the endothelial CX43-PARP1-NAD+ pathway's role in vascular aging and identify a potential therapeutic strategy to combat aging-associated BBB leakage with neuroprotective implications.


Assuntos
Conexina 43 , NAD , Animais , Humanos , Camundongos , Envelhecimento/fisiologia , Barreira Hematoencefálica/metabolismo , Encéfalo/metabolismo , Conexina 43/genética , Conexina 43/metabolismo , NAD/metabolismo , Poli(ADP-Ribose) Polimerase-1/metabolismo
19.
Biosens Bioelectron ; 238: 115562, 2023 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-37586262

RESUMO

Norfloxacin (NOR) residues in water pose a serious threat to human health via the food chain, necessitating the development of a rapid on-site antibiotic detection technique. In this work, we utilize electrostatic spinning technology that combines polyacrylonitrile (PAN) fibers and adenosine triphosphate (ATP)-rare earth metal Tb3+ complexes (ATP/Tb) to construct a new ternary film-based sensor for sensitive, quick, and convenient field testing of NOR in water. The operating mechanism is that the ternary system produces gradually enhanced bright green fluorescence at increasing concentrations of NOR. The unique fluorescence property of the ternary systems is attributed to the use of ATP, rather than the commonly used adenosine monophosphate (AMP), to coordinate with Tb3+, which avoided the possible fluorescence quenching from competitive water binding. Benefiting from the PAN nanofiber's superior stability, acid, and alkali resistance, and flexibility as support, the ternary system exhibited a good linear response to NOR in a wide dynamic range of 0.04-30 µM at the detection limit of 16 nM. Additionally, the combination of a smartphone color recognition app allows for quick on-scene NOR detection. This film sensing strategy is instructive for the development of smart and portable sensing platforms for real-time detection of analytes such as antibiotics, pesticide residues, and hazardous materials in water bodies.


Assuntos
Técnicas Biossensoriais , Nanofibras , Humanos , Norfloxacino , Espectrometria de Fluorescência/métodos , Antibacterianos , Trifosfato de Adenosina , Água , Corantes Fluorescentes/química , Limite de Detecção , Smartphone
20.
Cogn Neurodyn ; 17(3): 703-713, 2023 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-37265654

RESUMO

Epilepsy is a chronic disorder caused by excessive electrical discharges. Currently, clinical experts identify the seizure onset zone (SOZ) channel through visual judgment based on long-time intracranial electroencephalogram (iEEG), which is a very time-consuming, difficult and experience-based task. Therefore, there is a need for high-accuracy diagnostic aids to reduce the workload of clinical experts. In this article, we propose a method in which, the iEEG is split into the 20-s segment and for each patient, we ask clinical experts to label a part of the data, which is used to train a model and classify the remaining iEEG data. In recent years, machine learning methods have been successfully applied to solve some medical problems. Filtering, entropy and short-time Fourier transform (STFT) are used for extracting features. We compare them to wavelet transform (WT), empirical mode decomposition (EMD) and other traditional methods with the aim of obtaining the best possible discriminating features. Finally, we look for their medical interpretation, which is important for clinical experts. We achieve high-performance results for SOZ and non-SOZ data classification by using the labeled iEEG data and support vector machine (SVM), fully connected neural network (FCNN) and convolutional neural network (CNN) as classification models. In addition, we introduce the positive unlabeled (PU) learning to further reduce the workload of clinical experts. By using PU learning, we can learn a binary classifier with a small amount of labeled data and a large amount of unlabeled data. This can greatly reduce the amount and difficulty of annotation work by clinical experts. All together, we show that using 105 minutes of labeled data we achieve a classification result of 91.46% on average for multiple patients.

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