RESUMO
The advancement of spatial transcriptomics (ST) technology contributes to a more profound comprehension of the spatial properties of gene expression within tissues. However, due to challenges of high dimensionality, pronounced noise and dynamic limitations in ST data, the integration of gene expression and spatial information to accurately identify spatial domains remains challenging. This paper proposes a SpaNCMG algorithm for the purpose of achieving precise spatial domain description and localization based on a neighborhood-complementary mixed-view graph convolutional network. The algorithm enables better adaptation to ST data at different resolutions by integrating the local information from KNN and the global structure from r-radius into a complementary neighborhood graph. It also introduces an attention mechanism to achieve adaptive fusion of different reconstructed expressions, and utilizes KPCA method for dimensionality reduction. The application of SpaNCMG on five datasets from four sequencing platforms demonstrates superior performance to eight existing advanced methods. Specifically, the algorithm achieved highest ARI accuracies of 0.63 and 0.52 on the datasets of the human dorsolateral prefrontal cortex and mouse somatosensory cortex, respectively. It accurately identified the spatial locations of marker genes in the mouse olfactory bulb tissue and inferred the biological functions of different regions. When handling larger datasets such as mouse embryos, the SpaNCMG not only identified the main tissue structures but also explored unlabeled domains. Overall, the good generalization ability and scalability of SpaNCMG make it an outstanding tool for understanding tissue structure and disease mechanisms. Our codes are available at https://github.com/ZhihaoSi/SpaNCMG.
Assuntos
Algoritmos , Transcriptoma , Humanos , Animais , Camundongos , Perfilação da Expressão Gênica/métodos , Redes Neurais de Computação , Biologia Computacional/métodos , Córtex Pré-Frontal/metabolismoRESUMO
Verticillium wilt caused by the soil-borne fungus Verticillium dahliae Kleb., is a destructive plant disease that instigates severe losses in many crops. Improving plant resistance to Verticillium wilt has been a challenge in most crops. In this study, a V. dahliae secreted protein VdSP8 was identified and shown to activate hyper-sensitive response (HR) and systemic acquired resistance (SAR) to Pseudomonas syringae pv. tomato DC3000 (Pst DC3000) and Botrytis cinerea in tobacco plants. We identified a ß-glucosidase named GhBGLU46 as a cotton plant target of VdSP8. VdSP8 interacts with GhBGLU46 both in vivo and in vitro and promotes the ß-glucosidase activity of GhBGLU46. Silencing of GhBGLU46 reduced the expression of genes involved in lignin biosynthesis, such as GhCCR4, GhCCoAOMT2, GhCAD3 and GhCAD6, thus decreasing lignin deposition and increasing Verticillium wilt susceptibility. We have shown that GhBGLU46 is indispensable for the function of VdSP8 in plant resistance. These results suggest that plants have also evolved a strategy to exploit the invading effector protein VdSP8 to enhance plant resistance.
RESUMO
Verticillium dahliae is a soilborne phytopathogenic fungus causing Verticillium wilt on hundreds of plant species. Several sequenced genomes of V. dahliae are available, but functional characterization of most genes has just begun. Based on our previous comparison of the transcriptome from the wild-type and ΔVdCf2 strains, a significant upregulation of the gene cassette, Vd276-280, in the ΔVdCf2 strain was observed. In this study, the functional characterization of the Vd276-280 gene cassette was performed. Agrobacterium-mediated knockout of this gene cassette in V. dahliae significantly inhibited conidiation, melanized microsclerotium formation in the mutant strains, and their virulence towards cotton. Furthermore, deletion of individual genes in the Vd276-280 gene cassette identified that the disruption of VDAG_07276 and VDAG_07280 delayed microsclerotium formation, inhibited conidiation, and reduced virulence towards cotton. Our data suggest that VDAG_07276 and VDAG_07280 in the Vd276-280 gene cassette mainly act as positive regulators of development and virulence in V. dahliae.
RESUMO
In the traditional method for hyperspectral anomaly detection, spectral feature mapping is used to map hyperspectral data to a high-level feature space to make features more easily distinguishable between different features. However, the uncertainty in the mapping direction makes the mapped features ineffective in distinguishing anomalous targets from the background. To address this problem, a hyperspectral anomaly detection algorithm based on the spectral similarity variability feature (SSVF) is proposed. First, the high-dimensional similar neighborhoods are fused into similar features using AE networks, and then the SSVF are obtained using residual autoencoder. Finally, the final detection of SSVF was obtained using Reed and Xiaoli (RX) detectors. Compared with other comparison algorithms with the highest accuracy, the overall detection accuracy (AUCODP) of the SSVFRX algorithm is increased by 0.2106. The experimental results show that SSVF has great advantages in both highlighting anomalous targets and improving separability between different ground objects.
RESUMO
OBJECTIVES: Bloodstream infection (BSI) is one of the major causes of death in pediatric tumor patients. Blood samples are relatively easy to obtain and thus provide a ready source of infection-related biological markers for the prompt evaluation of infection risk. METHODS: A total of 259 pediatric tumor patients were included from May 2019 to March 2022. Patients were divided into BSI group (n=70) and control group (n=189). Clinical and biological data were collected using electronic medical records. Differences in biological markers between BSI group and control group and differences before and during infection in BSI group were analyzed. RESULTS: The infected group showed higher levels of procalcitonin (PCT) and hypersensitive C-reactive-protein (hsCRP), and lower prealbumin (PA) than the uninfected group. Area under the receiver-operating curve (ROC) curves (AUC) of PCT, hsCRP and NLR (absolute neutrophil count to the absolute lymphocyte count) were 0.756, 0.617 and 0.612. The AUC of other biomarkers was ≤0.6. In addition, PCT, hsCRP, NLR and fibrinogen (Fg) were significantly increased during infection, while PA and lymphocyte (LYM) were significantly decreased. Antibiotic resistant of Gram-positive bacteria to CHL, SXT, OXA and PEN was lower than that of Coagulase-negative Staphylococcus. Resistant of Gram-positive bacteria to CHL was lower, while to SXT was higher than that of Gram-negative bacteria. CONCLUSIONS: This study explored the utility of biomarkers to assist in diagnosis and found that the PCT had the greatest predictive value for infection in pediatric tumor patients with BSI. Additionally, the PCT, hsCRP, NLR, PA, LYM and Fg were changed by BSI.
Assuntos
Bacteriemia , Neoplasias , Sepse , Criança , Humanos , Pró-Calcitonina , Proteína C-Reativa/análise , Neutrófilos/metabolismo , Curva ROC , Bacteriemia/diagnóstico , Estudos Retrospectivos , Sepse/diagnóstico , Linfócitos/metabolismo , Biomarcadores , Neoplasias/complicações , Neoplasias/diagnósticoRESUMO
Type 2C protein phosphatases regulate various biological processes in eukaryotes. However, their functions in Verticillium dahliae have not been characterized. In this study, homologs VdPtc1, VdPtc3, VdPtc5, VdPtc6, and VdPtc7 were identified in V. dahliae on the basis of homologous comparison with those in Saccharomyces cerevisiae. VdPtc2 and VdPtc4 are missing in the genome of the V. dahliae XJ592 strain. VdPtc3 is the homolog of Ptc2, Ptc3, and Ptc4 proteins in S. cerevisiae, implying that VdPtc3 may play versatile functions in V. dahliae. VdPtc3 promoted conidium development, melanin, and microsclerotium formation in V. dahliae. The ΔVdPtc3 strains showed increased sensitivity to NaCl and sorbitol and augmented the phosphorylation of p38 mitogen-activated protein kinase homolog Hog1 induced by osmotic stress. Besides, the ΔVdPtc3 strains also showed milder Verticillium wilt symptom on cotton. Furthermore, VdPtc3 interacts with VdAtg1, which modulates melanin and microsclerotium formation, as well as pathogenicity.
Assuntos
Ascomicetos , Verticillium , Virulência , Saccharomyces cerevisiae , Melaninas/genética , Melaninas/metabolismo , Monoéster Fosfórico Hidrolases/metabolismo , Doenças das Plantas , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismoRESUMO
Verticillium wilt is primarily caused by the fungus Verticillium dahliae and represents one of the most important worldwide soilborne plant diseases. The causal agent can be spread by microsclerotia and conidia attached to seeds during national/international trade or in soil between fields. Consequently, accurate, sensitive, and rapid detection of V. dahliae from complex samples is critical for restricting entry of the pathogen to a new region/environment and enforcing early management of Verticillium wilt. Based on CRISPR/Cas12a and recombinant polymerase amplification (RPA) technologies, we developed an accurate, sensitive, and rapid detection method for V. dahliae with paper-based lateral flow strips. A highly efficient and specific CRISPR RNA (crRNA) was designed for the GAPDH gene of V. dahliae and was validated using several closely related Verticillium spp. Excluding the time required for the DNA extraction from the complex samples, a minimum of 40 min was required for the detection time. The RPA-CRISPR/Cas12a detection system had a lower detection limit of â¼10 copies of genomic DNA per reaction and was able to successfully detect as little as one microsclerotium per gram of soil. In addition, field samples displaying symptoms suggestive of V. dahliae were able to be positively identified for the presence of V. dahliae. Taken together, this study broadens the applications of CRISPR/Cas12a nucleic acid detection to soilborne crop diseases and will contribute to the future development of field-deployable diagnostic tools.
Assuntos
Ascomicetos , Sistemas CRISPR-Cas , Comércio , Internacionalidade , Solo , TecnologiaRESUMO
Seeds play a critical role in the production of American ginseng. Seeds are also one of the most important media for the long-distant dissemination and the crucial way for pathogen survival. Figuring out the pathogens carried by seeds is the basis for effective management of seedborne diseases. In this paper, we tested the fungi carried by the seeds of American ginseng from the main production areas of China using incubation and highly throughput sequencing methods. The seed-carried rates of fungi in Liuba, Fusong, Rongcheng, and Wendeng were 100, 93.8, 75.2, and 45.7%, respectively. Sixty-seven fungal species, which belonged to 28 genera, were isolated from the seeds. Eleven pathogens were identified from the seed samples. Among the pathogens, Fusarium spp. were found in all of the seed samples. The relative abundance of Fusarium spp. in the kernel was higher than that in the shell. Alpha index showed that the fungal diversity between seed shell and kernel differed significantly. Nonmetric multidimensional scaling analysis revealed that the samples from different provinces and between seed shell and kernel were distinctly separated. The inhibition rates of four fungicides to seed-carried fungi of American ginseng were 71.83% for Tebuconazole SC, 46.67% for Azoxystrobin SC, 46.08% for Fludioxonil WP, and 11.11% for Phenamacril SC. Fludioxonil, a conventional seed treatment agent, showed a low inhibitory effect on seed-carried fungi of American ginseng.
Assuntos
Fungicidas Industriais , Fusarium , Panax , Endófitos/genética , Fusarium/genética , Fungicidas Industriais/farmacologia , Sementes/microbiologia , Panax/microbiologiaRESUMO
A/J mouse is a typical animal model of age-related deafness. Previous studies have shown that the mice suffer from progressive hearing loss and degeneration of cochlear cells, and a variation of H55 N in citrate synthase (CS) causes about 40% the hearing loss. CS is a key enzyme in the tricarboxylic acid cycle, which is transported from cytoplasm to mitochondria after synthesis, sorted by the mitochondrial targeting sequence (MTS). To explore the mechanism of CS (H55 N) variation in affecting its function, HEI-OC1 cells were infected with lentivirus particles to express CS-Flag or CS(H55 N)-Flag. The results showed that H55 N variation in CS, as purified by co-immunoprecipitation, decreased the enzyme activity by about 50%. Confocal microscope co-localization indicated that the CS (H55 N) variation led to a decrement in its mitochondrial content. Western blot also showed the amount of CS(H55 N)-Flag was more than that of CS(WT)-Flag in the cytosol. The results suggest H55 N variation in CS lead to decrement of its enzyme activity and targeting transport to mitochondria. We therefore conclude that decrement in CS activity and mitochondrial delivery contributes to the degeneration of cochlear cells and thus the hearing loss in A/J mice.
Assuntos
Perda Auditiva , Mitocôndrias , Animais , Citrato (si)-Sintase , Cóclea , CamundongosRESUMO
Transcription factors (TFs) bind to the promoters of target genes to regulate gene expression in response to different stimuli. The functions and regulatory mechanisms of transcription factors (TFs) in Verticillium dahliae are, however, still largely unclear. This study showed that a C2H2-type zinc finger TF, VdCf2 (V. dahliae chorion transcription factor 2), plays key roles in V. dahliae growth, melanin production, and virulence. Transcriptome sequencing analysis showed that VdCf2 was involved in the regulation of expression of genes encoding secreted proteins, pathogen-host interaction (PHI) homologs, TFs, and G protein-coupled receptors (GPCRs). Furthermore, VdCf2 positively regulated the expression of VdPevD1 (VDAG_02735), a previously reported virulence factor. VdCf2 thus regulates the expression of several pathogenicity-related genes that also contribute to virulence in V. dahliae. VdCf2 also inhibited the transcription of the Vd276-280 gene cluster and interacted with two members encoding proteins (VDAG_07276 and VDAG_07278) in the gene cluster. IMPORTANCE Verticillium dahliae is an important soilborne phytopathogen which can ruinously attack numerous host plants and cause significant economic losses. Transcription factors (TFs) were reported to be involved in various biological processes, such as hyphal growth and virulence of pathogenic fungi. However, the functions and regulatory mechanisms of TFs in V. dahliae remain largely unclear. In this study, we identified a new transcription factor, VdCf2 (V. dahliae chorion transcription factor 2), based on previous transcriptome data, which participates in growth, melanin production, and virulence of V. dahliae. We provide evidence that VdCf2 regulates the expression of the pathogenicity-related gene VdPevD1 (VDAG_02735) and Vd276-280 gene cluster. VdCf2 also interacts with VDAG_07276 and VDAG_07278 in this gene cluster based on a yeast two-hybrid and bimolecular fluorescence complementation assay. These results revealed the regulatory mechanisms of a pivotal pathogenicity-related transcription factor, VdCf2 in V. dahliae.
Assuntos
Verticillium , Virulência/genética , Verticillium/genética , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Metabolismo Secundário , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Melaninas/genética , Fatores de Virulência/genética , Fatores de Virulência/metabolismo , Família Multigênica , Interações Hospedeiro-Patógeno , Doenças das Plantas/microbiologiaRESUMO
BACKGROUND: Citrate Synthase (Cs) gene mutation (locus ahL4) has been found to play an important role in progressive hearing loss of A/J mice. HEI-OC1 cells have been widely used as an in vitro system to study cellular and molecular mechanisms related to hearing lose. We previously reported the increased apoptosis and the accumulation of reactive oxygen species in shRNACs-1429 cells, a Cs low-expressed cell model from HEI-OCI. The details of the mechanism of ROS production and apoptosis mediated by the abnormal expression of Cs needed to research furtherly. METHODS: iTRAQ proteomics was utilized to detect the differentially expressed proteins (DEPs) caused by low expression of Cs. The GO and KEGG pathways analysis were performed for annotation of the differentially expressed proteins. Protein-protein interaction network was constructed by STRING online database. Immunoblotting was utilized to confirm the protein levels of the the differentially expressed proteins. RESULTS: The differentially expressed proteins were significantly enriched in various signaling pathways mainly related to mitochondrial dysfunction diseases including Parkinson's disease, Alzheimer's disease, Huntington's disease, et al. Most noteworthy, the oxidative phosphorylation pathway was most significantly suppressed in the shRNACs-1429 cells,, in which a total of 10 differentially expressed proteins were enriched and were all downregulated by the abnormal expression of Cs. The downregulations of Ndufb5, Ndufv1 and Uqcrb were confirmed by immunoblotting. Meanwhile, the ATP levels of shRNACs-1429 cells were also reduced. CONCLUSIONS: These results suggest that low level expression of Cs induces the inhibition of oxidative phosphorylation pathway, which is responsible for the high level production of reactive oxygen species and low level of ATP, leading to the apoptosis of cochlear cells. This study may provide new theories for understanding and therapy of progressive hearing loss.
RESUMO
SETDB1 is a histone lysine methyltransferase that has critical roles in cancers. However, its potential role in gastric cancer (GC) remains obscure. Here, we mainly investigate the clinical significance and the possible role of SETDB1 in GC. We find that SETDB1 expression is upregulated in GC tissues and its high-level expression was a predictor of poor prognosis in patients. Overexpression of SETDB1 promoted cell proliferation and metastasis, while SETDB1 suppression had an opposite effect both in vitro and in vivo. Mechanistically, SETDB1 was shown to interact with ERG to promote the transcription of cyclin D1 (CCND1) and matrix metalloproteinase 9 (MMP9) through binding to their promoter regions. In addition, the expression of SETDB1 was also enhanced by the transcription factor TCF4 at the transcriptional level in GC. Furthermore, SETDB1 expression was found to be induced by Helicobacter pylori (H. pylori) infection in a TCF4-dependent manner. Taken together, our results indicate that SETDB1 is aberrantly overexpressed in GC and plays key roles in gastric carcinogenesis and metastasis via upregulation of CCND1 and MMP9. Our work also suggests that SETDB1 could be a potential oncogenic factor and a therapeutic target for GC. © 2020 The Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.
Assuntos
Ciclina D1/metabolismo , Infecções por Helicobacter/patologia , Histona-Lisina N-Metiltransferase/metabolismo , Metaloproteinase 9 da Matriz/metabolismo , Neoplasias Gástricas/genética , Fator de Transcrição 4/metabolismo , Animais , Carcinogênese , Linhagem Celular Tumoral , Proliferação de Células , Ciclina D1/genética , Progressão da Doença , Feminino , Histona-Lisina N-Metiltransferase/genética , Humanos , Metaloproteinase 9 da Matriz/genética , Camundongos Endogâmicos BALB C , Invasividade Neoplásica , Metástase Neoplásica , Regiões Promotoras Genéticas/genética , Estômago/patologia , Neoplasias Gástricas/diagnóstico , Neoplasias Gástricas/patologia , Fator de Transcrição 4/genética , Regulação para CimaRESUMO
American ginseng (Panax quinquefolium L.) originating from North America is one of important herbal medicine and economic crops . With the increasing market demand, China has become the third producer and the largest consumer country of American ginseng. However, continuous cropping obstacle has become the most serious problem for the production of American ginseng, and the continuous cropping of soils usually lead to accumulations of root fungal pathogens and increasing plant disease occurrence (1), root rot caused by the notorious soil-borne pathogenic fungi, Fusarium spp., results in a significant reduction of yield and quality of American ginseng. Investigation of American ginseng root rot was carried out in Liuba county, Shaanxi province, China from 2017 to 2019. About 20% of over 3-year-old American ginseng showed varied root rot symptoms in newly reclaimed fields, and more than 70% in continuous American ginseng planting fields. Among these root rot diseases, we found one kind of disease which shows symptoms of red leaves in initial stage and yellow or yellow brown lesions at the reed heads or taproots. The lesions mainly appear on the root surface; however, the vascular tissue has no discoloration. The aboveground parts become wilted and died, and the whole root appears dark brown rots. Fifteen Fusarium spp. isolates were obtained by cutting diseased rot roots into 5 × 5 mm2 pieces, disinfecting in 70% ethanol for 1 min, rinsing 2 ~ 3 times in sterile water for 1 min and isolating on PDA medium including 50 µg/mL streptomycin sulfate. All the isolates have identical morphological characteristics. The colony was white with curved and uplifted aerial hyphae in central region. The colony diameter was 48 ~51 mm after 6 days at room temperature. Microconidia were oval to cylindrical shape with 0 to 1 septa, ranged from 6.24 to10.09 µm long; the macroconidia were fusiform to conical with a hooked apical cell and a foot-shaped basal cell, usually 3 to 5 septa, ranged from 31.45 to 42.52 µm long. The chlamydospores were not found under our culture condition. Preliminary data analysis showed that the morphological characteristics of these isolates were consistent with the descriptions of Fusarium redolens (2). To clarify the fungus in the taxonomy , the rDNA internal transcribed spacer (ITS), the translation elongation factor 1 alpha (TEF1-α) and the RNA polymerase II subunit 1 (RPB1) fragments of two randomly selected isolates were amplified and sequenced. The sequences of the corresponding fragments of the two isolates were identical. The blast results in the GenBank and FUSARIUM-ID databases show the isolates belong to F. redolens (3). Previous study indicated F. redolens has an indistinguishable relative, F. hostae (4). Although the ITS sequence (MW331695) cannot provide enough information to distinguish them, the phylogenetic tree combined the sequence of TEF1-α (tempID: 2407237 ) and RPB1 (tempID: 2407229) clearly showed that the isolates are F. redolens. (Fig) The pathogenicity of a representative isolate, YP04, was tested on ginseng taproot by in vivo inoculation experiments with three replications. The taproot surface of 2-year-old healthy ginseng was washed and disinfested with 75% alcohol for 1 min and rinsed with sterile water, and dried. The surface of taproot was injured with sterilized steel needles and immersed in 1 × 106 /ml spore suspension (sterile water for control plants) for 30 min. The treatment and control plants were transplanted in 20 cm diameter flowerpots filled with sterilized humus and cultured in a greenhouse at 18-23°C. Six days after transplanting, the leaves began to turn red. The cortex of ginseng taproot showed yellow brown lesions and the vascular tissue turn to light yellow. Fifteen days after transplanting, the aboveground parts of treatment plants began to wilting and the taproots showed serious rots. no taproot rot was observed in the controls. The pathogen was re-isolated from the diseased taproots successfully. To our knowledge, this is the first report of F. redolens causing root rot of American ginseng in China.
RESUMO
Verticillium dahliae is a widely distributed soilborne pathogen that causes vascular wilt in more than 200 plant species. Defoliating and nondefoliating symptoms caused by the disease that result in either the loss or retention of leaves in infected plants, respectively, in hosts such as cotton, olive, and okra, divide the causal agent into defoliating and nondefoliating pathotypes. Our goal in this current work was to generate genome resources for the defoliating strain XJ592 and the nondefoliating strain XJ511 of V. dahliae isolated from cotton in China.
Assuntos
Genoma Fúngico , Plantas , Verticillium , China , Genoma Fúngico/genética , Doenças das Plantas/microbiologia , Folhas de Planta/microbiologia , Plantas/microbiologia , Verticillium/genéticaRESUMO
BACKGROUND: The autosomal recessive non-syndromic deafness DFNB28 is characterized by prelingual sensorineural hearing loss. The disease is related with mutations in TRIOBP (Trio- and F-actin-Binding Protein) gene, which has three transcripts referred to as TRIOBP-5, TRIOBP - 4 and TRIOBP-1. Among them, TRIOBP-5/- 4 are expressed in the inner ears and crucial for maintaining the structure and function of the stereocilia. METHODS: The proband is a 26-year-old Chinese female. She and her younger brother have being suffered from severe deafness since birth, whereas her parents, who are cousins, have normal communication ability. Hearing impairment of the two siblings was determined by pure tone audiometry. Whole Exome Sequencing (WES) was performed on the genomic DNA of the proband and Sanger sequencing was conducted on the DNA samples of the four family members. RESULTS: Tests of pure tone hearing thresholds showed a severe to profound symmetric hearing loss for the proband and her younger brother. Moreover, a novel TRIOBP c.1342C > T (p.Arg448*) variant was identified by WES in the DNA sample of the proband and confirmed by Sanger sequencing in DNA of the family members. CONCLUSIONS: The TRIOBP c.1342C > T (p.Arg448*) variant is predicted to disrupt TRIOBP-5 and TRIOBP-4, which may lead to the congenital deafness. The results will broaden the spectrum of pathogenic variants in TRIOBP gene. The characteristics of deafness in the family imply that marriage between close relatives should be avoided.
Assuntos
Estudos de Associação Genética , Predisposição Genética para Doença , Perda Auditiva Neurossensorial/diagnóstico , Perda Auditiva Neurossensorial/genética , Proteínas dos Microfilamentos/genética , Mutação , Adulto , Povo Asiático/genética , Audiometria , Tronco Encefálico/metabolismo , Tronco Encefálico/fisiopatologia , Consanguinidade , Feminino , Humanos , Polimorfismo de Nucleotídeo Único , Análise de Sequência de DNA , Sequenciamento do ExomaRESUMO
A/J mouse is a model of age-related hearing loss (AHL). Mutation in the citrate synthase (Cs) gene of the mouse plays an important role in the hearing loss and degeneration of cochlear cells. To investigate the pathogenesis of cochlear cell damage in A/J mice resulted from Cs mutation, we downregulated the expression level of CS in HEI-OC1, a cell line of mouse cochlea, by shRNA. The results showed that low CS expression led to low ability of cell proliferation. Further study revealed an increase level of reactive oxygen species (ROS), activation of ATF6 mediated endoplasmic reticulum stress (ERS) and high expression levels of caspase12 and Bax in the cells. Moreover, the AEBSF, an ATF6 inhibitor, could reduce the expression levels of caspase-12 and Bax by inhibiting the hydrolysis of ATF6 in the cells. Finally, antioxidant alpha-lipoic acid (ALA) reduced the ROS levels and the apoptotic signals in the cell model with low CS expression. We therefore conclude that the ERS mediated apoptosis, which is triggered by ROS, may be involved in the cell degeneration in the cochleae of A/J mice.
Assuntos
Antioxidantes/farmacologia , Apoptose/efeitos dos fármacos , Citrato (si)-Sintase/metabolismo , Estresse do Retículo Endoplasmático/fisiologia , Estresse Oxidativo/efeitos dos fármacos , Ácido Tióctico/farmacologia , Fator 6 Ativador da Transcrição/antagonistas & inibidores , Animais , Apoptose/fisiologia , Caspase 12/metabolismo , Linhagem Celular , Proliferação de Células/fisiologia , Regulação para Baixo , Estresse do Retículo Endoplasmático/efeitos dos fármacos , Camundongos , Mitocôndrias/metabolismo , Modelos Biológicos , Estresse Oxidativo/fisiologia , Presbiacusia/fisiopatologia , Espécies Reativas de Oxigênio/metabolismo , Sulfonas/farmacologia , Proteína X Associada a bcl-2/metabolismoRESUMO
BACKGROUND: Previous studies have demonstrated that the expression of homeobox8 (HOXB8) is higher in colorectal cancer (CRC) tissues than in normal tissues; however, the precise role of HOXB8 in human CRC cells remains to be elucidated. METHODS: We generated lentiviral constructs to overexpress and silence HOXB8 in CRC cell lines, and examined their biological functions through MTT, wound healing, colony and transwell, expression of signal transducer and activator of transcription 3 (STAT3) and epithelial-mesenchymal transition (EMT) related factors through western-blot. RESULTS: HOXB8 knockdown inhibited cellular proliferation and invasion in vitro as well as carcinogenesis and metastasis in vivo. HOXB8 also induced EMT, which is characterized by the down-regulation of E-cadherin and the up-regulation of Vimentin, N-cadherin, Twist, Zeb1 and Zeb2. Moreover, HOXB8 activated STAT3, which is known to play an oncogenic role in diverse human malignancies. CONCLUSIONS: Our results indicate that HOXB8 may be an independent prognostic factor in CRC. Therefore, deserved a deeper research.
RESUMO
Verticillium wilt caused by Verticillium spp., also called potato early dying disease, is one of the most serious soilborne diseases affecting potato production in China. The disease has been expanding into most potato production areas over the past few years. Information on host resistance against Verticillium wilt among the potato cultivars in China is scarce, but it is critical for sustainable management of the disease. This study, therefore, evaluated 30 commercially popular potato cultivars against Verticillium dahliae strain Vdp83 and Verticillium nonalfalfae strain Vnp24, two well-characterized strains causing Verticillium wilt of potato in China. Both strains were isolated from diseased potato plants, and they were previously proven to be highly virulent. Ten plants of each cultivar were inoculated with the V. dahliae strain and incubated on greenhouse benches. Symptoms were rated at weekly intervals, and the relative area under the disease progress curve was calculated. The experiment was repeated once, and nonparametric analysis was used to calculate the relative marginal effects and the corresponding confidence intervals. Five resistant cultivars and four susceptible cultivars identified from the analyses were then challenged with the V. nonalfalfae strain. Cultivar responses to V. nonalfalfae were like those exhibited against V. dahliae, except for one cultivar. This study showed that resistance among potato cultivars exists in China against Verticillium spp. and that the resistance to V. dahliae identified in potato is also effective against the other Verticillium species that cause Verticillium wilt with a few exceptions. Cultivars identified as resistant to Verticillium wilt can be deployed to manage the disease until the breeding programs develop new cultivars with resistance from the sources identified in this study.
Assuntos
Resistência à Doença , Solanum tuberosum , Verticillium , China , Doenças das Plantas/microbiologia , Solanum tuberosum/microbiologia , Especificidade da Espécie , Verticillium/fisiologiaRESUMO
We previously reported a novel positive feedback loop between thioredoxin-1 (Trx-1) and S100P, which promotes the invasion and metastasis of colorectal cancer (CRC). However, the underlying molecular mechanisms remain poorly understood. In this study, we examined the roles of Trx-1 and S100P in CRC epithelial-to-mesenchymal transition (EMT) and their underlying mechanisms. We observed that knockdown of Trx-1 or S100P in SW620 cells inhibited EMT, whereas overexpression of Trx-1 or S100P in SW480 cells promoted EMT. Importantly, S100A4 and the phosphorylation of AKT were identified as potential downstream targets of Trx-1 and S100P in CRC cells. Silencing S100A4 or inhibition of AKT phosphorylation eliminated S100P- or Trx-1-mediated CRC cell EMT, migration and invasion. Moreover, inhibition of AKT activity reversed S100P- or Trx-1-induced S100A4 expression. The expression of S100A4 was higher in human CRC tissues compared with their normal counterpart tissues and was significantly correlated with lymph node metastasis and poor survival. The overexpression of S100A4 protein was also positively correlated with S100P or Trx-1 protein overexpression in our cohort of CRC tissues. In addition, overexpression of S100P reversed the Trx-1 knockdown-induced inhibition of S100A4 expression, EMT and migration and invasion in SW620 cells. The data suggest that interplay between Trx-1 and S100P promoted CRC EMT as well as migration and invasion by up-regulating S100A4 through AKT activation, thus providing further potential therapeutic targets for suppressing the EMT in metastatic CRC.
Assuntos
Proteínas de Ligação ao Cálcio/genética , Neoplasias Colorretais/genética , Proteínas de Neoplasias/genética , Proteína A4 de Ligação a Cálcio da Família S100/genética , Tiorredoxinas/genética , Linhagem Celular Tumoral , Movimento Celular/genética , Proliferação de Células/genética , Neoplasias Colorretais/patologia , Transição Epitelial-Mesenquimal/genética , Humanos , Invasividade Neoplásica/genética , Invasividade Neoplásica/patologia , Proteína Oncogênica v-akt/genética , Fosforilação , Proteínas Proto-Oncogênicas c-aktRESUMO
This article proposes a novel active localization method based on the mixed polarization multiple signal classification (MP-MUSIC) algorithm for positioning a metal target or an insulator target in the underwater environment by using a uniform circular antenna (UCA). The boundary element method (BEM) is introduced to analyze the boundary of the target by use of a matrix equation. In this method, an electric dipole source as a part of the locating system is set perpendicularly to the plane of the UCA. As a result, the UCA can only receive the induction field of the target. The potential of each electrode of the UCA is used as spatial-temporal localization data, and it does not need to obtain the field component in each direction compared with the conventional fields-based localization method, which can be easily implemented in practical engineering applications. A simulation model and a physical experiment are constructed. The simulation and the experiment results provide accurate positioning performance, with the help of verifying the effectiveness of the proposed localization method in underwater target locating.