RESUMEN
The computer is an important medium that allows people to connect to the internet. However, people with disabilities are unable to use a computer mouse and thus cannot enjoy internet benefits. Nowadays, there are various types of assistive technologies for controlling a computer mouse, but they all have some operational inconveniences. In this paper, we propose an innovative blowing-controlled mouse assistive tool to replace the conventional hand-controlled mouse. Its main contribution is that it uses microphones to induce small signals through the principle of airflow vibration, and it then converts the received signal into the corresponding pulse width. The co-design of software programming enables various mouse functions to be implemented by the identification of the blowing pulse width of multiple microphones. The proposed tool is evaluated experimentally, and the experimental results show that the average identification rate of the proposed mouse is over 85%. Additionally, compared with the other mouse assistive tools, the proposed mouse has the benefits of low cost and humanized operation. Therefore, the proposed blowing control method can not only improve the life quality of people with disabilities but also overcome the disadvantages of existing assistive tools.
RESUMEN
Bamboo mosaic virus (BaMV) satellite RNA (satBaMV) depends on BaMV for its replication and encapsidation. SatBaMV-encoded P20 protein is an RNA-binding protein that facilitates satBaMV systemic movement in co-infected plants. Here, we examined phosphorylation of P20 and its regulatory functions. Recombinant P20 (rP20) was phosphorylated by host cellular kinase(s) in vitro, and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and mutational analyses revealed Ser-11 as the phosphorylation site. The phosphor-mimic rP20 protein interactions with satBaMV-translated mutant P20 were affected. In overlay assay, the Asp mutation at S11 (S11D) completely abolished the self-interaction of rP20 and significantly inhibited the interaction with both the WT and S11A rP20. In chemical cross-linking assays, S11D failed to oligomerize. Electrophoretic mobility shift assay and subsequent Hill transformation analysis revealed a low affinity of the phospho-mimicking rP20 for satBaMV RNA. Substantial modulation of satBaMV RNA conformation upon interaction with nonphospho-mimic rP20 in circular dichroism analysis indicated formation of stable satBaMV ribonucleoprotein complexes. The dissimilar satBaMV translation regulation of the nonphospho- and phospho-mimic rP20 suggests that phosphorylation of P20 in the ribonucleoprotein complex converts the translation-incompetent satBaMV RNA to messenger RNA. The phospho-deficient or phospho-mimicking P20 mutant of satBaMV delayed the systemic spread of satBaMV in co-infected Nicotiana benthamiana with BaMV. Thus, satBaMV likely regulates the formation of satBaMV RNP complex during co-infection in planta.
Asunto(s)
Potexvirus/genética , Satélite de ARN/metabolismo , Proteínas de Unión al ARN/metabolismo , Proteínas Virales/metabolismo , Secuencia de Aminoácidos , Dimerización , Regulación hacia Abajo , Datos de Secuencia Molecular , Mutación , Conformación de Ácido Nucleico , Fosforilación , Biosíntesis de Proteínas , Satélite de ARN/química , Proteínas de Unión al ARN/química , Proteínas de Unión al ARN/genética , Ribonucleoproteínas/metabolismo , Serina/fisiología , Nicotiana/virología , Proteínas Virales/química , Proteínas Virales/genéticaRESUMEN
Satellite RNAs associated with Bamboo mosaic virus (satBaMVs) depend on BaMV for replication and encapsidation. Certain satBaMVs isolated from natural fields significantly interfere with BaMV replication. The 5' apical hairpin stem loop (AHSL) of satBaMV is the major determinant in interference with BaMV replication. In this study, by in vivo competition assay, we revealed that the sequence and structure of AHSL, along with specific nucleotides (C(60) and C(83)) required for interference with BaMV replication, are also involved in replication competition among satBaMV variants. Moreover, all of the 5' ends of natural BaMV isolates contain the similar AHSLs having conserved nucleotides (C(64) and C(86)) with those of interfering satBaMVs, suggesting their co-evolution. Mutational analyses revealed that C(86) was essential for BaMV replication, and that replacement of C(64) with U reduced replication efficiency. The non-interfering satBaMV interfered with BaMV replication with the BaMV-C64U mutant as helper. These findings suggest that two cytosines at the equivalent positions in the AHSLs of BaMV and satBaMV play a crucial role in replication competence. The downregulation level, which is dependent upon the molar ratio of interfering satBaMV to BaMV, implies that there is competition for limited replication machinery.
Asunto(s)
Potexvirus/genética , Satélite de ARN/química , Replicación Viral/genética , Citosina/química , Regulación hacia Abajo , Conformación de Ácido Nucleico , Potexvirus/fisiología , Protoplastos/virología , Nicotiana/virología , Regiones no TraducidasRESUMEN
BACKGROUND/AIM: Lenvatinib, an oral multikinase inhibitor, has demonstrated promising activity in patients with osteosarcoma (OS). Therefore, it is worth exploring the inhibitory efficacy and mechanism of action of lenvatinib in osteosarcoma. The primary goal of this study was to examine the inhibitory effectiveness and mechanism of lenvatinib on the growth and invasion of OS cells. MATERIALS AND METHODS: The effects of lenvatinib on cell viability, apoptosis, protein kinase B (AKT) activation, its downstream effector proteins involved in tumor progression, and invasion capability were assessed using MTT assay, flow cytometry, western blotting, and invasion/migration assay on U-2 OS and MG63 cells. RESULTS: Lenvatinib effectively induced cytotoxicity, apoptosis, as well as extrinsic and intrinsic apoptotic signaling in OS cells. Lenvatinib also significantly decreased the invasion/migration capability, AKT activation, and downstream effector proteins. CONCLUSION: The anti-OS effect of lenvatinib may be associated with the induction of apoptosis and the inactivation of AKT.
Asunto(s)
Neoplasias Óseas , Osteosarcoma , Humanos , Proteínas Proto-Oncogénicas c-akt/metabolismo , Línea Celular Tumoral , Proliferación Celular , Movimiento Celular , Apoptosis , Osteosarcoma/patología , Neoplasias Óseas/patologíaRESUMEN
This study evaluates the safety and effectiveness of computed tomography- (CT-) assisted endoscopic surgery in the treatment of infectious spondylodiscitis of the thoracic and upper lumbar spine in immunocompromised patients. From October 2006 to March 2014, a total of 41 patients with infectious spondylodiscitis underwent percutaneous endoscopic surgery under local anesthesia, and 13 lesions from 13 patients on the thoracic or upper lumbar spine were selected for evaluation. A CT-guided catheter was placed before percutaneous endoscopic surgery as a guide to avoid injury to visceral organs, major vessels, and the spinal cord. All 13 patients had quick pain relief after endoscopic surgery without complications. The bacterial culture rate was 77%. Inflammatory parameters returned to normal after adequate antibiotic treatment. Postoperative radiographs showed no significant kyphotic deformity when compared with preoperative films. As of the last follow-up visit, no recurrent infections were noted. Traditional transthoracic or diaphragmatic surgery with or without posterior instrumentation is associated with high rates of morbidity and mortality, especially in elderly patients, patients with multiple comorbidities, or immunocompromised patients. Percutaneous endoscopic surgery assisted by a CT-guided catheter provides a safe and effective alternative treatment for infectious spondylodiscitis of the thoracic and upper lumbar spine.
Asunto(s)
Discitis/inmunología , Discitis/cirugía , Endoscopía/métodos , Huésped Inmunocomprometido/inmunología , Vértebras Lumbares/cirugía , Procedimientos Quirúrgicos Mínimamente Invasivos/métodos , Anciano , Anciano de 80 o más Años , Infecciones Bacterianas/inmunología , Infecciones Bacterianas/patología , Infecciones Bacterianas/cirugía , Discitis/patología , Endoscopía/efectos adversos , Femenino , Humanos , Masculino , Persona de Mediana Edad , Procedimientos Quirúrgicos Mínimamente Invasivos/efectos adversos , Cirugía Asistida por Computador/métodosAsunto(s)
Fascitis Necrotizante/patología , Fracturas Abiertas/patología , Infecciones por Bacterias Gramnegativas/patología , Cirrosis Hepática Alcohólica/patología , Fracturas de la Tibia/patología , Adulto , Aeromonas hydrophila , Antibacterianos/uso terapéutico , Fascitis Necrotizante/diagnóstico por imagen , Fascitis Necrotizante/microbiología , Fascitis Necrotizante/cirugía , Resultado Fatal , Fracturas Abiertas/diagnóstico por imagen , Fracturas Abiertas/microbiología , Fracturas Abiertas/cirugía , Infecciones por Bacterias Gramnegativas/diagnóstico por imagen , Infecciones por Bacterias Gramnegativas/tratamiento farmacológico , Infecciones por Bacterias Gramnegativas/microbiología , Humanos , Cirrosis Hepática Alcohólica/diagnóstico por imagen , Cirrosis Hepática Alcohólica/microbiología , Masculino , Fracturas de la Tibia/diagnóstico por imagen , Fracturas de la Tibia/microbiología , Fracturas de la Tibia/cirugía , Tomografía Computarizada por Rayos XRESUMEN
Plant genetic engineering has broadened the options for plant virus resistance and is mostly based on pathogen-derived resistance. Previously, we have shown that interfering satellite RNA (satRNA) of Bamboo mosaic virus (satBaMV) greatly reduces Bamboo mosaic virus (BaMV) accumulation and BaMV-induced symptoms in co-inoculated plants. Here, we generated a nonviral source of virus-resistant transgenic Nicotiana benthamiana and Arabidopsis thaliana by introducing interfering satBaMV. Asymptomatic transgenic N. benthamiana lines were highly resistant to BaMV virion and viral RNA infection, and the expression of the transgene BSL6 was higher in asymptomatic than mildly symptomatic lines. In addition, BaMV- and satBaMV-specific small RNAs were detectable only after BaMV challenge, and their levels were associated with genomic viral RNA or satRNA levels. By transcriptomic analysis, the salicylic acid (SA) signalling pathway was not induced in satBaMV transgenic A. thaliana in mock conditions, suggesting that two major antiviral mechanisms, RNA silencing and SA-mediated resistance, are not involved directly in transgenic satBaMV-mediated BaMV interference. In contrast, resistance is associated with the level of the interfering satBaMV transgene. We propose satBaMV-mediated BaMV interference in transgenic plants by competition for replicase with BaMV.
Asunto(s)
Arabidopsis/virología , Resistencia a la Enfermedad/inmunología , Virus del Mosaico/fisiología , Nicotiana/virología , Enfermedades de las Plantas/inmunología , Enfermedades de las Plantas/virología , Satélite de ARN/metabolismo , Arabidopsis/genética , Resistencia a la Enfermedad/genética , Regulación Bacteriana de la Expresión Génica , Interacciones Huésped-Patógeno/genética , Modelos Biológicos , Fenotipo , Plantas Modificadas Genéticamente , Satélite de ARN/genética , ARN Viral/genética , ARN Viral/metabolismo , Nicotiana/genética , Transcriptoma/genética , Virión/metabolismo , Replicación ViralRESUMEN
Satellite RNAs associated with Bamboo mosaic virus (satBaMV) exhibit different phenotypes. Some isolates could reduce the accumulation of BaMV RNA and attenuate the BaMV-induced symptoms in co-inoculated plants. The determinants of the downregulation of BaMV replication were mapped in the 5' hypervariable region of satBaMV, which folds into a conserved apical hairpin stem loop (AHSL) structure comprising an apical loop and two internal loops, as evidenced by enzymatic probing. We also demonstrated that the integrity of the AHSL structure of interfering satBaMV was essential for the interference of BaMV accumulation. Concurrent analyses of natural satBaMV isolates revealed that all of the interfering isolates contained the same structures and sequences in the internal loops. Further, refined analyses indicated that, besides the AHSL structure, specific nucleotides in the internal loops play a crucial role in the downregulation, which implies that they may be required for the interaction of viral/cellular factors in this process.
Asunto(s)
Regiones no Traducidas 5'/fisiología , Regulación hacia Abajo , Enfermedades de las Plantas/virología , Potexvirus/fisiología , Satélite de ARN/fisiología , Replicación Viral/fisiología , Regiones no Traducidas 5'/genética , Secuencia de Bases , Modelos Moleculares , Datos de Secuencia Molecular , Conformación de Ácido Nucleico , Potexvirus/genética , Satélite de ARN/química , Satélite de ARN/genética , Nicotiana/virología , Replicación Viral/genéticaRESUMEN
Satellite RNA of Bamboo mosaic virus (satBaMV), a single-stranded mRNA type satellite encoding a protein of 20 kDa (P20), depends on the helper BaMV for replication and encapsidation. Two satBaMV isolates, BSF4 and BSL6, exhibit distinctly differential phenotypes in Nicotiana benthamiana plants when coinoculated with BaMV RNA. BSL6 significantly reduces BaMV RNA replication and suppresses the BaMV-induced symptoms, whereas BSF4 does not. By studies with chimeric satBaMVs generated by exchanging the components between BSF4 and BSL6, the genetic determinants responsible for the downregulation of BaMV replication and symptom expression were mapped at the 5' untranslated region (UTR) of BSL6. The 5' UTR of BSL6 alone is sufficient to diminish BaMV RNA replication when the 5' UTR is inserted in cis into the BaMV expression vector or when coinoculation with mutants that block the synthesis of P20 protein takes place. Further, the 5' UTR of natural satBaMV isolates contains one hypervariable (HV) region which folds into a conserved apical hairpin stem-loop (AHSL) structure (W. B. Yeh, Y. H. Hsu, H. C. Chen, and N. S. Lin, Virology 330:105-115, 2004). Interchanges of AHSL segment of HV regions between BSF4 and BSL6 led to the ability of chimeric satBaMV to interfere with BaMV replication and symptom expression. The conserved secondary structure within the HV region is a potent determinant of the downregulation of helper virus replication.
Asunto(s)
Regiones no Traducidas 5'/fisiología , Virus Helper/fisiología , Conformación de Ácido Nucleico , Potexvirus/fisiología , Satélite de ARN/genética , ARN Viral/genética , Replicación Viral/genética , Regiones no Traducidas 5'/genética , Secuencia de Bases , Regulación hacia Abajo , Modelos Moleculares , Datos de Secuencia Molecular , Enfermedades de las Plantas/virología , Potexvirus/genética , Satélite de ARN/fisiología , ARN Viral/fisiología , Nicotiana/virologíaRESUMEN
Satellite RNA (satRNA) associated with Bamboo mosaic virus (BaMV) is dependent on BaMV for replication and encapsidation. Molecular analyses of total RNA extracted from bamboo species collected worldwide revealed that 26 out of 61 BaMV isolates harbored satBaMV. Among them, two phylogenetically distinguishable groups, A and B, with a genetic diversity of 6.9 +/- 0.7% were identified. Greatest sequence diversity occurred in the 5' untranslated region (UTR) that contained one hypervariable region with variations of up to 20.7%. Concurrent covariations in the 5' hypervariable sequences support the existence of a conserved apical hairpin stem-loop structure, which was earlier mapped by enzymatic probings and functional analyses [Annamalai, P., Hsu, Y.H., Liu, Y.P., Tsai, C.H., Lin, N.S., 2003. Structural and mutational analyses of cis-acting sequences in the 5'-untranslated region of satellite RNA of bamboo mosaic potexvirus. Virology 311 (1), 229-239]. Furthermore, chimeric satBaMVs generated by interchanging the hypervariable region between groups A and B demonstrated the replication competence of satBaMV isolates in Nicotiana benthamiana protoplasts co-inoculated with BaMV RNA. The results suggest that an evolutionarily conserved secondary structure exists in the hypervariable region of 5' UTR of satBaMV.