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1.
Arch Microbiol ; 206(7): 292, 2024 Jun 07.
Artículo en Inglés | MEDLINE | ID: mdl-38849633

RESUMEN

In recent years, the evolution of antibiotic resistance has led to the inefficacy of several antibiotics, and the reverse of resistance was a novel method to solve this problem. We previously demonstrated that matrine (Mat) and berberine hydrochloride (Ber) had a synergistic effect against multidrug-resistant Escherichia coli (MDREC). This study aimed to demonstrate the effect of Mat combined with Ber in reversing the resistance of MDREC. The MDREC was sequenced passaged in the presence of Mat, Ber, and a combination of Mat and Ber, which did not affect its growth. The reverse rate was up to 39.67% after MDREC exposed to Mat + Ber for 15 days. The strain that reversed resistance was named drug resistance reversed E. coli (DRREC) and its resistance to ampicillin, streptomycin, gentamicin, and tetracycline was reversed. The MIC of Gentamicin Sulfate (GS) against DRREC decreased 128-fold to 0.63 µg/mL, and it was stable within 20 generations. Furthermore, the susceptible phenotype of DRREC remained stable within 20 generations, as well. The LD50 of DRREC for chickens was 8.69 × 109 CFU/mL. qRT-PCR assays revealed that the transcript levels of antibiotic-resistant genes and virulence genes in the DRREC strain were significantly lower than that in the MDREC strain (P < 0.05). In addition, GS decreased the death, decreased the bacterial loading in organs, alleviated the injury of the spleen and liver, and decreased the cytokine levels in the chickens infected by the DRREC strain. In contrast, the therapeutic effect of GS in chickens infected with MDREC was not as evident. These findings suggest that the combination of Mat and Ber has potential for reversing resistance to MDREC.


Asunto(s)
Alcaloides , Antibacterianos , Berberina , Pollos , Farmacorresistencia Bacteriana Múltiple , Infecciones por Escherichia coli , Escherichia coli , Gentamicinas , Matrinas , Pruebas de Sensibilidad Microbiana , Enfermedades de las Aves de Corral , Quinolizinas , Animales , Gentamicinas/farmacología , Escherichia coli/efectos de los fármacos , Escherichia coli/genética , Berberina/farmacología , Antibacterianos/farmacología , Quinolizinas/farmacología , Infecciones por Escherichia coli/veterinaria , Infecciones por Escherichia coli/tratamiento farmacológico , Infecciones por Escherichia coli/microbiología , Alcaloides/farmacología , Enfermedades de las Aves de Corral/microbiología , Enfermedades de las Aves de Corral/tratamiento farmacológico , Virulencia/efectos de los fármacos , Sinergismo Farmacológico
2.
Sensors (Basel) ; 24(6)2024 Mar 16.
Artículo en Inglés | MEDLINE | ID: mdl-38544170

RESUMEN

In recent years, the rapid prevalence of high-definition video in Internet of Things (IoT) systems has been directly facilitated by advances in imaging sensor technology. To adapt to limited uplink bandwidth, most media platforms opt to compress videos to bitrate streams for transmission. However, this compression often leads to significant texture loss and artifacts, which severely degrade the Quality of Experience (QoE). We propose a latent feature diffusion model (LFDM) for compressed video quality enhancement, which comprises a compact edge latent feature prior network (ELPN) and a conditional noise prediction network (CNPN). Specifically, we first pre-train ELPNet to construct a latent feature space that captures rich detail information for representing sharpness latent variables. Second, we incorporate these latent variables into the prediction network to iteratively guide the generation direction, thus resolving the problem that the direct application of diffusion models to temporal prediction disrupts inter-frame dependencies, thereby completing the modeling of temporal correlations. Lastly, we innovatively develop a Grouped Domain Fusion module that effectively addresses the challenges of diffusion distortion caused by naive cross-domain information fusion. Comparative experiments on the MFQEv2 benchmark validate our algorithm's superior performance in terms of both objective and subjective metrics. By integrating with codecs and image sensors, our method can provide higher video quality.

3.
Funct Integr Genomics ; 23(2): 99, 2023 Mar 23.
Artículo en Inglés | MEDLINE | ID: mdl-36959488

RESUMEN

Duck hepatitis A virus 1 (DHAV-1) is one of the main contagious pathogens that causes rapid death of ducklings. To illuminate the potential of DHAV-1-infected underlying mechanisms, we analyzed the mRNA and microRNA (miRNA) expression profiles of duck embryonic hepatocytes (DEHs) in response to DHAV-1. We found 3410 differentially expressed genes (DEGs) and 142 differentially expressed miRNAs (DEMs) at 36 h after DHAV-1 infection. Additionally, DEGs and the target genes of miRNA expression were analyzed and enriched utilizing GO and KEGG, which may be crucial for immune responses, viral resistance, and mitophagy. For instance, the dysregulation of DDX58, DHX58, IRF7, IFIH1, STING1, TRAF3, CALCOCO2, OPTN, PINK1, and MFN2 in DHAV-1-infected DEHs was verified by RT-qPCR. Then, the association analysis of mRNAs and miRNAs was constructed utilizing the protein-protein interaction (PPI) networks, and the expressions of main miRNAs were confirmed, including miR-132c-3p, miR-6542-3p, and novel-mir163. These findings reveal a synthetic characterization of the mRNA and miRNA in DHAV-1-infected DEHs and advance the understanding of molecular mechanism in DHAV-1 infection, which may provide a hint for the interactions of virus and host.


Asunto(s)
Virus de la Hepatitis del Pato , MicroARNs , Animales , Virus de la Hepatitis del Pato/genética , MicroARNs/genética , RNA-Seq , Patos/genética , ARN Mensajero/genética
4.
Plant Biotechnol J ; 21(5): 961-978, 2023 05.
Artículo en Inglés | MEDLINE | ID: mdl-36632704

RESUMEN

Despite the established significance of WRKY proteins and phenylpropanoid metabolism in plant immunity, how WRKY proteins modulate aspects of the phenylpropanoid pathway remains undetermined. To understand better the role of WRKY proteins in plant defence, we identified a cotton (Gossypium hirsutum) protein, GhWRKY41, that is, universally and rapidly induced in three disease-resistant cotton cultivars following inoculation with the plant pathogenic fungus, Verticillium dahliae. We show that overexpression of GhWRKY41 in transgenic cotton and Arabidopsis enhances resistance to V. dahliae, while knock-down increases cotton more susceptibility to the fungus. GhWRKY41 physically interacts with itself and directly activates its own transcription. A genome-wide chromatin immunoprecipitation and high-throughput sequencing (ChIP-seq), in combination with RNA sequencing (RNA-seq) analyses, revealed that 43.1% of GhWRKY41-binding genes were up-regulated in cotton upon inoculation with V. dahliae, including several phenylpropanoid metabolism master switches, receptor kinases, and disease resistance-related proteins. We also show that GhWRKY41 homodimer directly activates the expression of GhC4H and Gh4CL, thereby modulating the accumulation of lignin and flavonoids. This finding expands our understanding of WRKY-WRKY protein interactions and provides important insights into the regulation of the phenylpropanoid pathway in plant immune responses by a WRKY protein.


Asunto(s)
Ascomicetos , Verticillium , Gossypium/metabolismo , Retroalimentación , Proteínas de Plantas/metabolismo , Resistencia a la Enfermedad/genética , Enfermedades de las Plantas/microbiología , Regulación de la Expresión Génica de las Plantas
5.
Cancer Sci ; 113(1): 308-318, 2022 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-34710947

RESUMEN

Fusion of RET with different partner genes has been detected in papillary thyroid, lung, colorectal, pancreatic, and breast cancer. Approval of selpercatinib for treatment of lung and thyroid cancer with RET gene mutations or fusions calls for studies to explore RET fusion partners and their eligibility for RET-based targeted therapy. In this study, RET fusion patterns in a large group of Chinese cancer patients covering several cancer types were identified using next-generation sequencing. A total of 44 fusion patterns were identified in the study cohort with KIF5B, CCDC6, and ERC1 being the most common RET fusion partners. Notably, 17 novel fusions were first reported in this study. Prevalence of functional RET fusions was 1.05% in lung cancer, 6.03% in thyroid cancer, 0.39% in colorectal cancer, and less than 0.1% in gastric cancer and hepatocellular carcinoma. Analysis showed a preference for fusion partners in different tumor types, with KIF5B being the common type in lung cancer, CCDC6 in thyroid cancer, and NCOA4 in colorectal cancer. Co-occurrence of EGFR mutations and RET fusions with rare partner genes (rather than KIF5B) in lung cancer patients was correlated with epidermal growth factor receptor-tyrosine kinase inhibitor resistance and could predict response to targeted therapies. Findings from this study provide a guide to clinicians in determining tumors with specific fusion patterns as candidates for RET targeted therapies.


Asunto(s)
Pueblo Asiatico/genética , Perfilación de la Expresión Génica/métodos , Neoplasias/genética , Proteínas de Fusión Oncogénica/genética , Proteínas Proto-Oncogénicas c-ret/genética , Análisis de Secuencia de ADN/métodos , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , China , Femenino , Regulación Neoplásica de la Expresión Génica , Secuenciación de Nucleótidos de Alto Rendimiento , Humanos , Masculino , Persona de Mediana Edad , Prevalencia , Estudios Retrospectivos , Análisis de Secuencia de ARN , Adulto Joven
6.
Plant Biotechnol J ; 20(4): 691-710, 2022 04.
Artículo en Inglés | MEDLINE | ID: mdl-34800075

RESUMEN

Sea Island cotton (Gossypium barbadense) is the source of the world's finest fibre quality cotton, yet relatively little is understood about genetic variations among diverse germplasms, genes underlying important traits and the effects of pedigree selection. Here, we resequenced 336 G. barbadense accessions and identified 16 million SNPs. Phylogenetic and population structure analyses revealed two major gene pools and a third admixed subgroup derived from geographical dissemination and interbreeding. We conducted a genome-wide association study (GWAS) of 15 traits including fibre quality, yield, disease resistance, maturity and plant architecture. The highest number of associated loci was for fibre quality, followed by disease resistance and yield. Using gene expression analyses and VIGS transgenic experiments, we confirmed the roles of five candidate genes regulating four key traits, that is disease resistance, fibre length, fibre strength and lint percentage. Geographical and temporal considerations demonstrated selection for the superior fibre quality (fibre length and fibre strength), and high lint percentage in improving G. barbadense in China. Pedigree selection breeding increased Fusarium wilt disease resistance and separately improved fibre quality and yield. Our work provides a foundation for understanding genomic variation and selective breeding of Sea Island cotton.


Asunto(s)
Fusarium , Gossypium , Mapeo Cromosómico , Fibra de Algodón , Resistencia a la Enfermedad/genética , Genoma de Planta/genética , Estudio de Asociación del Genoma Completo , Gossypium/genética , Fenotipo , Filogenia , Fitomejoramiento , Sitios de Carácter Cuantitativo
7.
Int J Mol Sci ; 23(21)2022 Nov 03.
Artículo en Inglés | MEDLINE | ID: mdl-36362251

RESUMEN

Pollen grains, the male gametophytes for reproduction in higher plants, are vulnerable to various stresses that lead to loss of viability and eventually crop yield. A conventional method for assessing pollen viability is manual counting after staining, which is laborious and hinders high-throughput screening. We developed an automatic detection tool (PollenDetect) to distinguish viable and nonviable pollen based on the YOLOv5 neural network, which is adjusted to adapt to the small target detection task. Compared with manual work, PollenDetect significantly reduced detection time (from approximately 3 min to 1 s for each image). Meanwhile, PollenDetect can maintain high detection accuracy. When PollenDetect was tested on cotton pollen viability, 99% accuracy was achieved. Furthermore, the results obtained using PollenDetect show that high temperature weakened cotton pollen viability, which is highly similar to the pollen viability results obtained using 2,3,5-triphenyltetrazolium formazan quantification. PollenDetect is an open-source software that can be further trained to count different types of pollen for research purposes. Thus, PollenDetect is a rapid and accurate system for recognizing pollen viability status, and is important for screening stress-resistant crop varieties for the identification of pollen viability and stress resistance genes during genetic breeding research.


Asunto(s)
Aprendizaje Profundo , Fitomejoramiento , Polen , Programas Informáticos , Calor
8.
Int J Mol Sci ; 23(6)2022 Mar 21.
Artículo en Inglés | MEDLINE | ID: mdl-35328797

RESUMEN

Anther indehiscence and pollen sterility caused by high temperature (HT) stress have become a major problem that decreases the yield of cotton. Pollen- and anther-specific genes play a critical role in the process of male reproduction and the response to HT stress. In order to identify pollen-specific genes that respond to HT stress, a comparative transcriptome profiling analysis was performed in the pollen and anthers of Gossypium hirsutum HT-sensitive Line H05 against other tissue types under normal temperature (NT) conditions, and the analysis of a differentially expressed gene was conducted in the pollen of H05 under NT and HT conditions. In total, we identified 1111 pollen-specific genes (PSGs), 1066 anther-specific genes (ASGs), and 833 pollen differentially expressed genes (DEGs). Moreover, we found that the late stage of anther included more anther- and pollen-specific genes (APSGs). Stress-related cis-regulatory elements (CREs) and hormone-responsive CREs are enriched in the promoters of APSGs, suggesting that APSGs may respond to HT stress. However, 833 pollen DEGs had only 10 common genes with 1111 PSGs, indicating that PSGs are mainly involved in the processes of pollen development and do not respond to HT stress. Promoters of these 10 common genes are enriched for stress-related CREs and MeJA-responsive CREs, suggesting that these 10 common genes are involved in the process of pollen development while responding to HT stress. This study provides a pathway for rapidly identifying cotton pollen-specific genes that respond to HT stress.


Asunto(s)
Regulación de la Expresión Génica de las Plantas , Gossypium , Flores/metabolismo , Perfilación de la Expresión Génica , Gossypium/metabolismo , Polen/genética , Temperatura , Transcriptoma
9.
Molecules ; 26(14)2021 Jul 15.
Artículo en Inglés | MEDLINE | ID: mdl-34299563

RESUMEN

A new terthiophene-based imidazole luminophore 5,5'-(1H-thieno[3,4-d]imidazole-4,6-diyl)bis(thiophene-2-carboxylic acid) (TIBTCH2, 5) was synthesized in one step from previously reported 4,6-di(thiophen-2-yl)-1H-thieno[3,4-d]imidazole (DTTI, 4), and their photophysical properties were studied and compared accordingly. Under solvothermal conditions, reacting 5 with Mn(OAc)2 yielded a new three-dimensional metal-organic framework (MOF, 6) which was structurally defined by single-crystal X-ray diffraction. In 6, all Mn(II) ions octahedrally bind to carboxylate-O atoms to form a linear Mn3 secondary building unit (SBU) that contains three distinct coordination modes. Importantly, 6 exhibits dual functional properties of ligand-based emission and metal-based magnetic behaviors.

10.
Protein Expr Purif ; 175: 105688, 2020 11.
Artículo en Inglés | MEDLINE | ID: mdl-32681953

RESUMEN

BST2 is a single-pass type II transmembrane (TM) protein, which has a cytoplasmic domain, a transmembrane domain, and an extracellular domain, each domain is important for biologic function of BST2. BST2 is a host restriction factor that can effectively inhibit retrovirus release. Rhesus monkeys are considered as relevant natural animal models for studying AIDS in humans. In order to recognize rhesus BST2 (RhBST2) protein and detect its function accurately, we prepared a polyclonal antibody (pAb) especially for RhBST2. Meanwhile, we constructed RhBST2 proteins with the addition of an HA-tag at the N-terminus (RhBST2-NHA) or inside of the ectodomain (RhBST2-IHA) to compare the recognition ability of rabbit anti-RhBST2 pAb and anti-HA mAb. The results showed that the anti-HA mAb and rabbit anti-RhBST2 pAb had the same ability to identify RhBST2. RhBST2 demonstrated antiviral activity and the ability to activate NF-κB. Moreover, the N-glycosylation states, cell surface level and intracellular localization of RhBST2 were detected. However, HA tags relatively changed part of the biological function of RhBST2. These results show that the RhBST2 polyclonal antibody is more suitable for analyzing the properties and functions of RhBST2, and the natural domain of RhBST2 is very important for its function.


Asunto(s)
Anticuerpos/inmunología , Antígenos CD , Antivirales , Síndrome de Inmunodeficiencia Adquirida , Animales , Antígenos CD/biosíntesis , Antígenos CD/química , Antígenos CD/inmunología , Antivirales/química , Antivirales/inmunología , Antivirales/metabolismo , Proteínas Ligadas a GPI/biosíntesis , Proteínas Ligadas a GPI/química , Proteínas Ligadas a GPI/inmunología , Células HEK293 , VIH-1/inmunología , Humanos , Macaca mulatta , Dominios Proteicos , Conejos
11.
BMC Evol Biol ; 19(1): 134, 2019 06 26.
Artículo en Inglés | MEDLINE | ID: mdl-31242851

RESUMEN

BACKGROUND: Current patterns of population genetic variation may have been shaped by long-term evolutionary history and contemporary demographic processes. Understanding the underlying mechanisms that yield those patterns is crucial for informed conservation of endangered species. The critically endangered white-headed langur, Trachypithecus leucocephalus, is endemic to a narrow range in southwest China. This species shows very low genetic diversity in its 2 main relict populations, Fusui and Chongzuo. Whether this has been caused by a short evolutionary history or recent population declines is unknown. Therefore, we investigated the contributions of historical and recent population demographic changes to population genetic diversity by using 15 nuclear microsatellite markers and mitochondrial DNA (mtDNA) control region sequences. RESULTS: Using genetic data from 214 individuals we found a total of 9 mtDNA haplotypes in the Fusui population but only 1 haplotype in the Chongzuo population, and we found an overall low genetic diversity (haplotype and nucleotide diversities: h = 0.486 ± 0.036; π = 0.0028 ± 0.0003). The demographic history inferred from mtDNA and microsatellite markers revealed no evidence for historical population size fluctuations or recent population bottlenecks. Simulations of possible population divergence histories inferred by DIYABC analysis supported a recent divergence of the Chongzuo population from the Fusui population and no population bottlenecks. CONCLUSIONS: Despite severe population declines caused by anthropogenic activities in the last century, the low genetic diversity of the extant white-headed langur populations is most likely primarily due to the species' shallow evolutionary history and to a recent, local population founder event.


Asunto(s)
Colobinae/genética , Especies en Peligro de Extinción , Variación Genética , Animales , China , ADN Mitocondrial/genética , Haplotipos , Repeticiones de Microsatélite/genética , Densidad de Población
12.
Anal Chem ; 90(1): 708-715, 2018 01 02.
Artículo en Inglés | MEDLINE | ID: mdl-29202232

RESUMEN

In this study, a whole-course nucleic-acid-constructed biosensor that combines the all-in-one concepts of the universal blocking linker recombinase polymerase amplification (UBLRPA) and a peptide nucleic acid (PNA)-based lateral flow device (PLFD) has been developed for the ultrasensitive detection of food pathogens. Using the preamplification UBLRPA principle, a universal linker and C3 space blocker were utilized to produce the universal linker single-duplex DNA products. The developed amplification system was employed to convert duplex products to a single strand. In the signal recognition strategy, a special PNA probe was successfully employed in the portable PLFD. The UBLRPA products were identified visually using the PLFD through dual hybridization (a PNA probe on the gold nanoparticle (Au-NP) was combined with a universal linker on the end of the products; a PNA capture probe was used on the test line and a universal linker on the other end of the products). The accumulation of Au-NPs produced a characteristic red band, enabling the visual detection of a food pathogen without further testing. To demonstrate the value of the all-in-one biosensor, Salmonella enterica subsp. enterica serovar typhimurium was used as a model organism. The biosensor showed high selectivity and extraordinary repeatability using S. typhimurium, and the limit of detection was 4 CFU mL-1. Furthermore, when milk samples artificially contaminated with S. typhimurium were analyzed, the analysis was completed within 30 min without complicated instrumentation. The results exhibited good precision and recovery. This portable all-in-one biosensor demonstrates great promise for the screening of pathogens in food and environmental samples.


Asunto(s)
Técnicas Biosensibles/métodos , Microbiología de Alimentos/métodos , Ácidos Nucleicos de Péptidos/genética , Salmonella typhimurium/aislamiento & purificación , Animales , ADN/genética , Oro/química , Límite de Detección , Nanopartículas del Metal/química , Leche/microbiología , Técnicas de Amplificación de Ácido Nucleico/métodos , Hibridación de Ácido Nucleico
13.
BMC Plant Biol ; 18(1): 286, 2018 Nov 20.
Artículo en Inglés | MEDLINE | ID: mdl-30458710

RESUMEN

BACKGROUND: Plant architecture and the vegetative-reproductive transition have major impacts on the agronomic success of crop plants, but genetic mechanisms underlying these traits in cotton (Gossypium spp.) have not been identified. RESULTS: We identify four natural mutations in GoCEN-Dt associated with cluster fruiting (cl) and early maturity. The situ hybridization shows that GhCEN is preferentially expressed in cotton shoot apical meristems (SAM) of the main stem and axillary buds. Constitutive GhCEN-Dt overexpression suppresses the transition of the cotton vegetative apex to a reproductive shoot. Silencing GoCEN leads to early flowering and determinate growth, and in tetraploids causes the main stem to terminate in a floral bud, a novel phenotype that exemplifies co-adaptation of polyploid subgenomes and suggests new research and/or crop improvement approaches. Natural cl variations are enriched in cottons adapted to high latitudes with short frost-free periods, indicating that mutants of GoCEN have been strongly selected for early maturity. CONCLUSION: We show that the cotton gene GoCEN-Dt, a homolog of Antirrhinum CENTRORADIALIS, is responsible for determinate growth habit and cluster fruiting. Insight into the genetic control of branch and flower differentiation offers new approaches to develop early maturing cultivars of cotton and other crops with plant architecture appropriate for mechanical harvesting.


Asunto(s)
Genes de Plantas , Variación Genética , Gossypium/genética , Flores/genética , Frutas/crecimiento & desarrollo , Expresión Génica , Gossypium/crecimiento & desarrollo , Mutación , Fitomejoramiento
14.
Am J Primatol ; 79(5)2017 05.
Artículo en Inglés | MEDLINE | ID: mdl-28100010

RESUMEN

Dispersal is a critically important life history trait of social organisms that has a major impact on the population genetic structure and social relationships within groups. Primates exhibit highly diversified dispersal and philopatry patterns, but knowledge of these patterns is difficult to obtain and usually limited to observations of a small number of focal social groups or individuals. Here, we investigated the dispersal pattern of a critically endangered colobine monkey, the white-headed langur (Trachypithecus leucocephalus), using molecular approaches, and sex-specific population genetic structure analyses at fine geographical scales. We non-invasively collected 403 fecal samples from 41 social groups across 90% of the langur's range in Fusui (FS) and Chongzuo (CZ) in southwestern Guangxi Province, China. We identified 214 unique individuals from the samples by genotyping 15 polymorphic autosomal microsatellite loci, a sex-specific marker, and sequencing the mitochondrial DNA (mtDNA) hypervariable region I (HVRI). We found higher intragroup than intergroup genetic relatedness in males and females in both populations. A significant positive correlation between genetic distance and geographical distance, that is a pattern of isolation-by-distance, was detected in females from the FS population, but not in males. Spatial autocorrelation analyses revealed high within-group relatedness in both sexes and populations, as well as an additional positive correlation at the 0.5-km distance class in females from the FS population. Furthermore, we inferred first-generation migrants using genetic assignment tests. Our results suggest that male T. leucocephalus disperse at random distances within habitat areas, whereas dispersal of females may mainly occur among adjacent groups near their home site. Our study provides the first genetic evidence for sex-biased dispersal in T. leucocephalus, which has important management and conservation implications for the species.


Asunto(s)
Distribución Animal , Colobinae/fisiología , Genotipo , Repeticiones de Microsatélite , Conducta Social , Animales , China , Colobinae/genética , ADN Mitocondrial/genética , Especies en Peligro de Extinción , Femenino , Masculino , Análisis de Secuencia de ADN , Factores Sexuales
15.
Biochem Biophys Res Commun ; 461(4): 624-9, 2015 Jun 12.
Artículo en Inglés | MEDLINE | ID: mdl-25912140

RESUMEN

Rbx1 and Rbx2 are essential components of Cullin-RING E3 Ligases. Vif is generally believed to preferentially recruit the Cul5-Rbx2 module to induce proteasomal degradation of antiretroviral enzyme APOBEC3G, although some investigators have found that the Cul5-Rbx1 module is recruited. Here, to investigate the function of the two Rbx proteins in the Vif-Cul5 complex, we analyzed the performance of Cul5-Rbx1/Cul5-Rbx2 module in the activity of Vif E3 ligase and evaluated the interactions between Rbx1/Rbx2 and Cul5. We found that either Rbx1 or Rbx2 could promote ubiquitination of APOBE3G (A3G) in vitro. We also found that both Rbx1 and Rbx2 could bind Cul5 in cells and Rbx2 could dose-dependently inhibit the interaction of Rbx1 with Cul5. Furthermore, only the decrease of endogenous Rbx2 but not Rbx1 could impair the Vif-induced A3G degradation in cells. These findings indicate that Rbx1 and Rbx2 can both activate Cul5-Vif E3 ligase in vitro, but they may undergo a more delicate selection mechanism in vivo.


Asunto(s)
Proteínas Portadoras/química , Proteínas Portadoras/metabolismo , VIH-1/enzimología , Ubiquitina-Proteína Ligasas/química , Ubiquitina-Proteína Ligasas/metabolismo , Células HEK293 , Humanos , Complejos Multienzimáticos/química , Complejos Multienzimáticos/metabolismo , Ubiquitinación/fisiología , Productos del Gen vif del Virus de la Inmunodeficiencia Humana/química , Productos del Gen vif del Virus de la Inmunodeficiencia Humana/metabolismo
16.
Retrovirology ; 11: 77, 2014 Sep 12.
Artículo en Inglés | MEDLINE | ID: mdl-25213124

RESUMEN

BACKGROUND: All lentiviruses except equine infectious anemia virus (EIVA) antagonize antiviral family APOBEC3 (A3) proteins of the host through viral Vif proteins. The mechanism by which Vif of human, simian or feline immunodeficiency viruses (HIV/SIV/FIV) suppresses the corresponding host A3s has been studied extensively. RESULTS: Here, we determined that bovine immunodeficiency virus (BIV) and maedi-visna virus (MVV) Vif proteins utilize the Cullin (Cul)-ElonginB (EloB)-ElonginC (EloC) complex (BIV Vif recruits Cul2, while MVV Vif recruits Cul5) to degrade Bos taurus (bt)A3Z2-Z3 and Ovis aries (oa)A3Z2-Z3, respectively, via a proteasome-dependent but a CBF-ß-independent pathway. Mutation of the BC box in BIV and MVV Vif, C-terminal hydrophilic replacement of btEloC and oaEloC and dominant-negative mutants of btCul2 and oaCul5 could disrupt the activity of BIV and MVV Vif, respectively. While the membrane-permeable zinc chelator TPEN could block BIV Vif-mediated degradation of btA3Z2-Z3, it had minimal effects on oaA3Z2-Z3 degradation induced by MVV Vif, indicating that Zn is important for the activity of BIV Vif but not MVV Vif. Furthermore, we identified a previously unreported zinc binding loop [C-x1-C-x1-H-x19-C] in the BIV Vif upstream BC box which is critical for its degradation activity. CONCLUSIONS: A novel zinc binding loop was identified in the BIV Vif protein that is important for the E3 ubiquination activity, suggesting that the degradation of btA3Z2-Z3 by BIV and that of oaA3Z2-Z3 by MVV Vif may need host factors other than CBF-ß.


Asunto(s)
Proteínas Cullin/fisiología , Citosina Desaminasa/metabolismo , Productos del Gen vif/fisiología , Interacciones Huésped-Patógeno , Virus de la Inmunodeficiencia Bovina/fisiología , Factores de Transcripción/fisiología , Ubiquitina-Proteína Ligasas/fisiología , Virus Visna-Maedi/fisiología , Desaminasas APOBEC , Animales , Citidina Desaminasa , Elonguina , Productos del Gen vif/química , Células HEK293 , Humanos , Complejo de la Endopetidasa Proteasomal/fisiología
17.
Mol Vis ; 20: 1048-56, 2014.
Artículo en Inglés | MEDLINE | ID: mdl-25053875

RESUMEN

PURPOSE: MiRNAs are small noncoding RNAs that have been implicated in tumor development. They regulate target gene expression either by mRNA degradation or by translation repression. Activation of ß-catenin has been linked to pterygium progression. Here, we hypothesize that ß-catenin-associated miRNA, miRNA-221, and downstream p27Kip1 gene expression are correlated with the pathogenesis of pterygium. METHODS: We collected 120 pterygial and 120 normal conjunctival samples for this study. Immunohistochemistry and real-time reverse transcription (RT)-PCR were performed to determine ß-catenin protein localization, miR-221, and p27Kip1 gene expression. Pterygium cell line (PECs) cell models were used to confirm the effect of ß-catenin, miR-221, and p27Kip1 gene in the proliferation of pterygium cells. RESULTS: Seventy-two (60.0%) pterygial specimens showed high miR-221 expression levels, which was significantly higher than the control groups (13 of 120, 10.8%, p<0.0001). MiR-221 expression was significantly higher in ß-catenin-nuclear/cytoplasmic-positive groups than in ß-catenin membrane-positive and negative groups (p=0.001). We also found that p27Kip1 gene expression in pterygium was negatively correlated with miR-221 expression (p=0.002). In the clinical association, miR-221 expression was significantly higher in the fleshy and intermediate groups than in the atrophic group (p=0.007). The association of miR-221, p27Kip1 and proliferation of pterygium were also confirmed in the PECs model. CONCLUSIONS: Our study demonstrated that activation of ß-catenin in pterygium may interact with miR-221, resulting in p27Kip1 gene downregulation that influences pterygium pathogenesis.


Asunto(s)
Inhibidor p27 de las Quinasas Dependientes de la Ciclina/genética , Regulación de la Expresión Génica , MicroARNs/genética , Pterigion/etiología , Pterigion/genética , Anciano , Anciano de 80 o más Años , Línea Celular , Proliferación Celular , Conjuntiva/metabolismo , Conjuntiva/patología , Inhibidor p27 de las Quinasas Dependientes de la Ciclina/metabolismo , Femenino , Humanos , Masculino , MicroARNs/metabolismo , Persona de Mediana Edad , Modelos Biológicos , Transporte de Proteínas , Pterigion/patología , ARN Mensajero/genética , ARN Mensajero/metabolismo , Reacción en Cadena en Tiempo Real de la Polimerasa , beta Catenina/metabolismo
18.
Poult Sci ; 103(7): 103839, 2024 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-38810565

RESUMEN

Duck hepatitis A virus 1 (DHAV-1) is the primary cause of duck viral hepatitis, leading to sudden mortality in ducklings and significant economic losses in the duck industry. However, little is known about how DHAV-1 affects duckling liver at the molecular level. We conducted an analysis comparing the expression patterns of mRNAs and miRNAs in DHAV-1-infected duckling livers to understand the underlying mechanisms and dynamic changes. We identified 6,818 differentially expressed mRNAs (DEGs) and 144 differentially expressed microRNAs (DEMs) during DHAV-1 infection. Functional enrichment analysis of DEGs and miRNA target genes using gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) revealed their potential involvement in innate antiviral immunity, mitophagy, and pyroptosis. We constructed coexpression networks of mRNA-miRNA interactions and confirmed key DEMs (novel-mir333, novel-mir288, novel-mir197, and novel-mir71) using RT-qPCR. Further investigation demonstrated that DHAV-1 activates the RLRs signaling pathway, disrupts mitophagy, and induces pyroptosis. In conclusion, DHAV-1-induced antiviral immunity is closely linked to mitophagy, suggesting it could be a promising therapeutic target.


Asunto(s)
Patos , Virus de la Hepatitis del Pato , Hepatitis Viral Animal , MicroARNs , Mitofagia , Enfermedades de las Aves de Corral , ARN Mensajero , Transducción de Señal , Animales , Patos/genética , MicroARNs/genética , MicroARNs/metabolismo , Enfermedades de las Aves de Corral/virología , Enfermedades de las Aves de Corral/genética , Enfermedades de las Aves de Corral/inmunología , Virus de la Hepatitis del Pato/fisiología , Hepatitis Viral Animal/virología , Hepatitis Viral Animal/genética , Hepatitis Viral Animal/inmunología , ARN Mensajero/genética , ARN Mensajero/metabolismo , Infecciones por Picornaviridae/veterinaria , Infecciones por Picornaviridae/virología , Infecciones por Picornaviridae/inmunología , Infecciones por Picornaviridae/genética , Transcriptoma , Inmunidad Innata/genética
19.
Sci Total Environ ; 922: 171409, 2024 Apr 20.
Artículo en Inglés | MEDLINE | ID: mdl-38432367

RESUMEN

Arsenic (As) is a widespread metalloid with well-known toxicity. To date, numerous studies have focused on individual level toxicity (e.g., growth and reproduction) of As to typical invertebrate springtails in soils, however, the molecular level toxicity and mechanism was poorly understood. Here, an integrated transcriptomics and metabolomics approach was used to reveal responses of Folsomia candida exposed to As(V) of 10 and 60 mg kg-1 at which the individual level endpoints were influenced. Transcriptomics identified 5349 and 4020 differentially expressed genes (DEGs) in low and high concentration groups, respectively, and the most DEGs were down-regulated. Enrichment analysis showed that low and high concentrations of As(V) significantly inhibited chromatin/chromosome-related biological processes (chromatin/chromosome organization, nucleosome assembly and organization, etc.) in springtails. At high concentration treatment, structural constituent of cuticle, chitin metabolic process and peptidase activity (serine-type peptidase activity, endopeptidase activity, etc.) were inhibited or disturbed. Moreover, the apoptosis pathway was significantly induced. Metabolomics analysis identified 271 differential changed metabolites (DCMs) in springtails exposed to high concentration of As. Steroid hormone biosynthesis was the most significantly affected pathway. Several DCMs that related to chitin metabolism could further support above transcriptomic results. These findings further extended the knowledge of As toxic mechanisms to soil fauna and offer important information for the environmental risk assessment.


Asunto(s)
Arsénico , Artrópodos , Contaminantes del Suelo , Animales , Arsénico/metabolismo , Contaminantes del Suelo/metabolismo , Perfilación de la Expresión Génica , Metabolómica , Cromatina/metabolismo , Péptido Hidrolasas/metabolismo , Quitina/metabolismo , Suelo/química
20.
Res Vet Sci ; 170: 105178, 2024 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-38402660

RESUMEN

In pet clinics, the number of cases using trauma drugs accounts for >10% of the total number of cases, and most wounds are healing by second intention. The prolongation of wound healing time causes inconvenience and burden to pets and pet owners. Therefore, how to reduce wound healing time and achieve maximum recovery of tissue function and aesthetics is one of the focuses of veterinary clinical practice. Wound suppuration caused by Staphylococcus aureus and Pseudomonas aeruginosa is the main cause of delaying wound healing. Clinically, available antimicrobial treatments are almost exhausted due to the production of large numbers of resistant bacteria. At present, there are no bacteria resistant to traditional Chinese medicine (TCM), which makes TCM have the potential to become an effective drug for the treatment of bacterial infections, so the use of TCM in the treatment of traumatic infections has broad prospects. Based on the characteristics of infection syndrome, three different prescriptions were formulated in our laboratory, and the most effective prescription and dosage form was screened and named Lianrong Healing Cream (LRHC). The results showed that LRHC regulated the expression of fibroblast growth factor-2 (FGF-2), epidermal growth factor-1 (EGF-1), transforming growth factor-ß (TGF-ß) and vascular endothelial growth factor-1 (VEGF-1) genes in wound tissues and fibroblasts, thereby accelerating wound healing and repairing wound appearance and function. The results of this study may be help to develop TCM formulation for traumatic infections.


Asunto(s)
Medicina Tradicional China , Cicatrización de Heridas , Animales , Factor A de Crecimiento Endotelial Vascular/metabolismo , Factor de Crecimiento Epidérmico/farmacología
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