RESUMEN
This study investigated hepatitis B virus (HBV) single-nucleotide variants (SNVs) and deletion mutations linked with hepatocellular carcinoma (HCC). Ninety-three HCC patients and 108 non-HCC patients were enrolled for HBV genome-wide next-generation sequencing (NGS) analysis. A systematic literature review and a meta-analysis were performed to validate NGS-defined HCC-associated SNVs and deletions. The experimental results identified 60 NGS-defined HCC-associated SNVs, including 41 novel SNVs, and their pathogenic frequencies. Each SNV was specific for either genotype B (n = 24) or genotype C (n = 34), except for nt53C, which was present in both genotypes. The pathogenic frequencies of these HCC-associated SNVs showed a distinct U-shaped distribution pattern. According to the meta-analysis and literature review, 167 HBV variants from 109 publications were categorized into four levels (A-D) of supporting evidence that they are associated with HCC. The proportion of NGS-defined HCC-associated SNVs among these HBV variants declined significantly from 75% of 12 HCC-associated variants by meta-analysis (Level A) to 0% of 10 HCC-unassociated variants by meta-analysis (Level D) (P < 0.0001). PreS deletions were significantly associated with HCC, in terms of deletion index, for both genotypes B (P = 0.030) and C (P = 0.049). For genotype C, preS deletions involving a specific fragment (nt2977-3013) were significantly associated with HCC (HCC versus non-HCC, 6/34 versus 0/32, P = 0.025). Meta-analysis of preS deletions showed significant association with HCC (summary odds ratio 3.0; 95% confidence interval 2.3-3.9). Transfection of Huh7 cells showed that all of the five novel NGS-defined HCC-associated SNVs in the small surface region influenced hepatocarcinogenesis pathways, including endoplasmic reticulum-stress and DNA repair systems, as shown by microarray, real-time polymerase chain reaction and western blot analysis. Their carcinogenic mechanisms are worthy of further research. Copyright © 2017 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.
Asunto(s)
Carcinoma Hepatocelular/genética , Eliminación de Gen , Genoma Viral/genética , Virus de la Hepatitis B/genética , Neoplasias Hepáticas/genética , Polimorfismo de Nucleótido Simple/genética , Reparación del ADN/genética , Estrés del Retículo Endoplásmico/genética , Hepatitis B Crónica/genética , Humanos , Proteínas de Neoplasias/genética , Estudios RetrospectivosRESUMEN
This study proposes a home care system (HCS) based on a brain-computer interface (BCI) with a smartphone. The HCS provides daily help to motor-disabled people when a caregiver is not present. The aim of the study is two-fold: (1) to develop a BCI-based home care system to help end-users control their household appliances, and (2) to assess whether the architecture of the HCS is easy for motor-disabled people to use. A motion-strip is used to evoke event-related potentials (ERPs) in the brain of the user, and the system immediately processes these potentials to decode the user's intentions. The system, then, translates these intentions into application commands and sends them via Bluetooth to the user's smartphone to make an emergency call or to execute the corresponding app to emit an infrared (IR) signal to control a household appliance. Fifteen healthy and seven motor-disabled subjects (including the one with ALS) participated in the experiment. The average online accuracy was 81.8% and 78.1%, respectively. Using component N2P3 to discriminate targets from nontargets can increase the efficiency of the system. Results showed that the system allows end-users to use smartphone apps as long as they are using their brain waves. More important, only one electrode O1 is required to measure EEG signals, giving the system good practical usability. The HCS can, thus, improve the autonomy and self-reliance of its end-users.
Asunto(s)
Interfaces Cerebro-Computador , Servicios de Atención de Salud a Domicilio , Electroencefalografía , Potenciales Evocados , Humanos , Teléfono Inteligente , Interfaz Usuario-ComputadorRESUMEN
OBJECTIVES: Eight genotypes (A-H) of hepatitis B virus (HBV) are known with variations in nucleotide sequences greater than 8%. Several recent publications found that the clinical course and outcome of antiviral therapy depended on the genotype of the infecting HBV strain. Large epidemiological studies will require the availability of a system which is rapid, reliable and can be performed on a large number of samples. METHODS: To establish a simple and accurate genotyping method, the study collected 369 HBV complete genomic sequences from the GenBank database. Type-specific primers were also designed that separated HBV genotypes A to G by multiplex polymerase chain reaction. RESULTS: By comparison with the traditional restriction fragment length polymorphism method, over 93% of 441 samples were accurately genotyped by current assay, with a higher detection rate and sensitivity to detect mixed HBV infections. CONCLUSIONS: This methodology can be applied only to areas prevalent with HBV genotypes A to G. However, it provides an efficient alternative for clinical diagnosis and large-scale studies.
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Virus de la Hepatitis B/clasificación , Virus de la Hepatitis B/genética , Hepatitis B/virología , Reacción en Cadena de la Polimerasa/métodos , Cartilla de ADN/genética , Humanos , Sensibilidad y EspecificidadRESUMEN
Several hepatitis B virus (HBV) subgenotypes, HBV/A1, A2, Bj and Ba, have been reported with respect to clinical differences among patients infected with these subgenotypes. The population genetics and phylogeography of HBV were investigated based on the complete genome sequences of 484 isolates with 108 from our chronic hepatitis B patients and the remaining from the GenBank database. Besides genotypes A-H (HBV/A-H), five subgenotypes were identified among 169 HBV/B isolates by phylogenetic analysis and nucleotide divergence. There were 27 isolates of subgenotype B(1) (HBV/B(1)) restricted to Japan, 104 isolates of HBV/B(2) with the widest distribution in most Asian countries, 4 isolates of HBV/B(3) restricted to Indonesia, 32 isolates of HBV/B(4) restricted to Vietnam, and 7 isolates of HBV/B(5) restricted to Philippines. HBV/B(2)-B(5) isolates carried a recombination with HBV/C over the precore and core genes. In addition to the characteristics of HBV/B(1)-B(5) at some cis-acting elements, the precore stop-codon mutant (G1896A) was significantly different among HBV/B(1), HBV/B(2), and HBV/B(4) (70.3%, 31.7%, 53.0%, P=0.001), while no such mutation was found in HBV/B(3) and B(5). Among characteristics of the HBV/B(1)-B(5) amino acid sequences, serotype adw (K(122)) was exclusive among HBV/B(1), HBV/B(2), and HB V/B(3) isolates, while serotype ayw (R(122)) was among the HBV/B(4) and HBV/B(5) isolates. Furthermore, distinct variations of T cell and B cell recognition epitopes within surface and core proteins were also found among these subgenotypes. In conclusion, subgenotypes HBV/B(1)-B(5) exhibited distinct geographical distributions, virologic characteristics, and probable clinical implications.
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Genoma Viral , Virus de la Hepatitis B/clasificación , Virus de la Hepatitis B/genética , Hepatitis B Crónica/virología , Filogenia , Secuencia de Aminoácidos , Secuencia de Bases , Variación Genética , Genotipo , Hepacivirus/genética , Humanos , Epidemiología Molecular , Datos de Secuencia Molecular , Recombinación Genética , Proteínas Virales/química , Proteínas Virales/genéticaRESUMEN
This research analyzed the brain activity of non-native English speakers while engaged in English reading tests. The brain wave event-related potentials (ERPs) of participants were used to analyze the difference between making correct and incorrect choices on English reading test items. Three English reading tests of differing levels were designed and 20 participants, 10 males and 10 females whose ages ranged from 20 to 24, voluntarily participated in the experiment. Experimental results were analyzed by performing independent t-tests on the ERPs of participants for gender, difficulty level, and correct versus wrong options. Participants who chose incorrect options elicited a larger N600, verifying results found in the literature. Another interesting result was found: For incorrectly answered items, different areas of brain showing a significant difference in ERPs between the chosen and non-chosen options corresponded to gender differences; for males, this area was located in the right hemisphere whereas for females, it was located in the left. Experimental results imply that non-native English speaking males and females employ different areas of the brain to comprehend the meaning of difficult items.
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Encéfalo/fisiología , Multilingüismo , Lectura , Comprensión/fisiología , Electroencefalografía , Potenciales Evocados , Retroalimentación Psicológica/fisiología , Femenino , Humanos , Pruebas del Lenguaje , Masculino , Pruebas Neuropsicológicas , Reconocimiento Visual de Modelos/fisiología , Caracteres Sexuales , Adulto JovenRESUMEN
Molecular covariation of highly polymorphic viruses is thought to have crucial effects on viral replication and fitness. This study employs association rule data mining of hepatitis C virus (HCV) sequences to search for specific evolutionary covariation and then tests functional relevance on HCV replication. Data mining is performed between nucleotides in the untranslated regions 5' and 3'UTR, and the amino acid residues in the non-structural proteins NS2, NS3 and NS5B. Results indicate covariance of the 243(rd) nucleotide of the 5'UTR with the 14(th), 41(st), 76(th), 110(th), 211(th) and 212(th) residues of NS2 and with the 71(st), 175(th) and 621(st) residues of NS3. Real-time experiments using an HCV subgenomic system to quantify viral replication confirm replication regulation for each covariant pair between 5'UTR243 and NS2-41, -76, -110, -211, and NS3-71, -175. The HCV subgenomic system with/without the NS2 region shows that regulatory effects vanish without NS2, so replicative modulation mediated by HCV 5'UTR243 depends on NS2. Strong binding of the NS2 variants to HCV RNA correlates with reduced HCV replication whereas weak binding correlates with restoration of HCV replication efficiency, as determined by RNA-protein immunoprecipitation assay band intensity. The dominant haplotype 5'UTR243-NS2-41-76-110-211-NS3-71-175 differs according to the HCV genotype: G-Ile-Ile-Ile-Gly-Ile-Met for genotype 1b and A-Leu-Val-Leu-Ser-Val-Leu for genotypes 1a, 2a and 2b. In conclusion, 5'UTR243 co-varies with specific NS2/3 protein amino acid residues, which may have significant structural and functional consequences for HCV replication. This unreported mechanism involving HCV replication possibly can be exploited in the development of advanced anti-HCV medication.
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Regiones no Traducidas 5'/genética , Aminoácidos , Biología Computacional , Variación Genética/genética , Hepacivirus/genética , Nucleótidos/genética , Proteínas no Estructurales Virales/genética , Algoritmos , Línea Celular Tumoral , Secuencia Conservada , Minería de Datos , Genoma Viral/genética , Genotipo , Hepacivirus/fisiología , Humanos , Mutación , ARN Viral/genética , ARN Viral/metabolismo , Replicón/genética , Programas Informáticos , Proteínas no Estructurales Virales/química , Proteínas no Estructurales Virales/metabolismo , Replicación Viral/genéticaRESUMEN
Both the viral titer and the genotype significantly determine clinical outcomes and responses to antiviral treatment in chronic hepatitis B virus (HBV) infection. A method was developed for large-scale A-to-G genotyping with simultaneous viral quantification. The assay was run on a LightCycler instrument using hybridization probes. The genotype was determined from the melting points of the probes in a two-step manner. Set 1 amplicons differentiated genotypes B, E, and F from A, C, D, and G and simultaneously quantified viremia by real-time PCR. Melting curve analysis using the set 2-1 amplicon or the set 2-2 amplicon reaction mixture was then used to differentiate these genotype groups into single genotypes. HBV DNA quantification was consistent with that of the Amplicor assay and linear in a range from 10(2) to 10(13) copies/ml. By comparison with the restriction fragment length polymorphism method, 92.3% of 441 samples were accurately genotyped by the current assay. The method should be useful for genotyping and quantification of HBV DNA in areas where all genotypes exist.