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| 1 | Characterization of hepatitis B virus X gene quasispecies complexity in mono-infection and hepatitis delta virus superinfection显示文摘Hepatitis delta virus(HDV) seems to strongly suppress hepatitis B virus(HBV)replication, although little is known about the mechanism of this interaction. Both these viruses show a dynamic distribution of mutants, resulting in viral quasispecies. Next-generation sequencing is a viable approach for analyzing the composition of these mutant spectra. As the regulatory hepatitis B X protein(HBx) is essential for HBV replication, determination of HBV X gene(HBX)quasispecies complexity in HBV/HDV infection compared to HBV monoinfection may provide information on the interactions between these two viruses.AIM To compare HBV quasispecies complexity in the HBX 5' region between chronic hepatitis delta(CHD) and chronic HBV mono-infected patients.METHODS Twenty-four untreated patients were included: 7/24(29.2%) with HBeAgnegative chronic HBV infection(CI, previously termed inactive carriers), 8/24(33.3%) with HBeAg-negative chronic hepatitis B(CHB) and 9/24(37.5%) with CHD. A serum sample from each patient was first tested for HBV DNA levels.The HBX 5' region [nucleotides(nt) 1255-1611] was then PCR-amplified for subsequent next-generation sequencing(MiSeq, Illumina, United States). HBV quasispecies complexity in the region analyzed was evaluated using incidencebased indices(number of haplotypes and number of mutations), abundancebased indices(Hill numbers of order 1 and 2), and functional indices(mutation frequency and nucleotide diversity). We also evaluated the pattern of nucleotide changes to investigate which of them could be the cause of the quasispecies complexity.RESULTS CHB patients showed higher median HBV-DNA levels [5.4 logIU/mL,interquartile range(IQR) 3.5-7.9] than CHD(3.4 logIU/mL, IQR 3-7.6)(P = n.s.)or CI(3.2 logIU/mL, IQR 2.3-3.5)(P < 0.01) patients. The incidence and abundance indices indicated that HBV quasispecies complexity was significantly greater in CI than CHB. A similar trend was observed in CHD patients, although only Hill numbers of order 2 showed statistically significant differences(CHB2.81, IQR 1.11-4.57 vs CHD 8.87, 6.56-11.18, P = 0.038). There were no significant differences in the functional indices, but CI and CHD patients also showed a trend towards greater complexity than CHB. No differences were found for any HBV quasispecies complexity indices between CHD and CI patients. G-to-A and C-to-T nucleotide changes, characteristic of APOBEC3 G, were higher in CHD and CI than in CHB in genotype A haplotypes, but not in genotype D. The proportion of nt G-to-A vs A-to-G changes and C-to-T vs T-to-C changes in genotype A and D haplotypes in CHD patients showed no significant differences. In CHB and CI the results of these comparisons were dependent on HBV genotype.CONCLUSION The lower-replication CHD and CI groups show a trend to higher quasispecies complexity than the higher-replication CHB group. The mechanisms associated with this greater complexity require elucidation. | Cristina Godoy David Tabernero Sara Sopena Josep Gregori Maria Francesca Cortese Carolina González Rosario Casillas Mar?al Yll Ariadna Rando Rosa López-Martínez Josep Quer Gloria González-Aseguinolaza Rafael Esteban Mar Riveiro-Barciela Maria Buti Francisco Rodríguez-Frías | 2019 | World Journal of Gastroenterology2019，25，13： | 6 |
| 2 | Detection of hyper-conserved regions in hepatitis B virus X gene potentially useful for gene therapy显示文摘AIM To detect hyper-conserved regions in the hepatitis B virus(HBV) X gene(HBX) 5' region that could be candidates for gene therapy.METHODS The study included 27 chronic hepatitis B treatmentnaive patients in various clinical stages(from chronic infection to cirrhosis and hepatocellular carcinoma, both HBeA g-negative and HBeA g-positive), and infected with HBV genotypes A-F and H. In a serum sample from each patient with viremia > 3.5 log IU/m L, the HBX 5' end region [nucleotide(nt) 1255-1611] was PCRamplified and submitted to next-generation sequencing(NGS). We assessed genotype variants by phylogenetic analysis, and evaluated conservation of this region by calculating the information content of each nucleotide position in a multiple alignment of all unique sequences(haplotypes) obtained by NGS. Conservation at the HBx protein amino acid(aa) level was also analyzed.RESULTS NGS yielded 1333069 sequences from the 27 samples, with a median of 4578 sequences/sample(2487-9279, IQR 2817). In 14/27 patients(51.8%), phylogenetic analysis of viral nucleotide haplotypes showed a complex mixture of genotypic variants. Analysis of the information content in the haplotype multiple alignments detected 2 hyper-conserved nucleotide regions, one in the HBX upstream non-coding region(nt 1255-1286) and the other in the 5' end coding region(nt 1519-1603). This last region coded for a conserved amino acid region(aa 63-76) that partially overlaps a Kunitz-like domain.CONCLUSION Two hyper-conserved regions detected in the HBX 5' end may be of value for targeted gene therapy, regardless of the patients' clinical stage or HBV genotype. | Carolina González David Tabernero Maria Francesca Cortese Josep Gregori Rosario Casillas Mar Riveiro-Barciela Cristina Godoy Sara Sopena Ariadna Rando Marcal Yll Rosa Lopez-Martinez Josep Quer Rafael Esteban Maria Buti Francisco Rodríguez-Frías | 2018 | World Journal of Gastroenterology2018，24，19： | 6 |
| 3 | Analysis of hepatitis B virus preS1 variability and prevalence of the rs2296651 polymorphism in a Spanish population显示文摘AIM To determine the variability/conservation of the domain of hepatitis B virus(HBV) pre S1 region that interacts with sodium-taurocholate cotransporting polypeptide(hereafter, NTCP-interacting domain) and the prevalence of the rs2296651 polymorphism(S267 F, NTCP variant) in a Spanish population. METHODS Serum samples from 246 individuals were included and divided into 3 groups: patients with chronic HBV infection(CHB)(n = 41, 73% Caucasians), patients with resolved HBV infection(n = 100, 100% Caucasians) and an HBV-uninfected control group(n = 105, 100% Caucasians). Variability/conservation of the amino acid(aa) sequences of the NTCPinteracting domain,(aa 2-48 in viral genotype D) and a highly conserved pre S1 domain associated with virion morphogenesis(aa 92-103 in viral genotype D) were analyzed by next-generation sequencing and compared in 18 CHB patients with viremia > 4 log IU/mL. The rs2296651 polymorphism was determined in all individuals in all 3 groups using an in-house real-time PCR melting curve analysis.RESULTS The HBV pre S1 NTCP-interacting domain showed a high degree of conservation among the examined viral genomes especially between aa 9 and 21(in the genotype D consensus sequence). As compared with the virion morphogenesis domain, the NTCPinteracting domain had a smaller proportion of HBV genotype-unrelated changes comprising > 1% of the quasispecies(25.5% vs 31.8%), but a larger proportion of genotype-associated viral polymorphisms(34% vs 27.3%), according to consensus sequences from Gen Bank patterns of HBV genotypes A to H. Variation/conservation in both domains depended on viral genotype, with genotype C being the most highly conserved and genotype E the most variable(limited finding, only 2 genotype E included). Of note, proline residues were highly conserved in both domains, and serine residues showed changes only to threonine or tyrosine in the virion morphogenesis domain. The rs2296651 polymorphism was not detected in any participant.CONCLUSION In our CHB population, the NTCP-interacting domain was highly conserved, particularly the proline residues and essential amino acids related with the NTCP interaction, and the prevalence of rs2296651 was low/null. | Rosario Casillas David Tabernero Josep Gregori Irene Belmonte Maria Francesca Cortese Carolina González Mar Riveiro-Barciela Rosa Maria López Josep Quer Rafael Esteban Maria Buti Francisco Rodríguez-Frías | 2018 | World Journal of Gastroenterology2018，24，6： | 5 |
| 4 | Correlation between clinico-pathological outcome and typing of Haemophilus parasuis field strains显示文摘 | Virginia Aragon Marta Cerdà-Cuéllar Lorenzo Fraile Mark Mombarg Miquel Nofrarías Alexandre Olvera Marina Sibila David Solanes Joaquim Segalés | 2009 | Veterinary Microbiology2009，，3： | 2 |
| 5 | Semantic organizationand verbal memory efficiency in patients with schizophrenia显示文摘 | Brebion G David AS Jones H | 2004 | Neuropsychology2004，18，2： | 1 |
| 6 | Characterization of OXA-25, OXA-26, and OXA-27, Molecular Class Dβ-Lactamases Associated with Carbapenem Resistance in Clinical Isolates of Acinetobacter baumannii显示文摘 | Mariya AS Woodford N David ML | 2001 | Antimierobial Agents and Chemotherapy2001，45，58： | 1 |
| 7 | Receptor activated bladder andspinal ATP release in neutrally intact and chronic spinal cord in-jured rats显示文摘 | Nilson As alas GT David A | 2007 | Neurochen Int2007，50，： | 1 |
| 8 | Caring attributes, professional self concept and technological influences in a sample of registered nurses in eleven countries 显示文摘 | David AS Pang T Wong MF | 1999 | International Journal of Nursing Studies1999，36，： | 1 |
| 9 | Insight and psychiatric显示文摘 | David AS | 1990 | Br J Psychiatry1990，156，： | 1 |
| 10 | Insight and psychosis显示文摘 | David AS | 1990 | Br J Psychaictry1990，156，6： | 1 |
| 11 | The behavior and emotional correlates of epilepsy in adolescence : a 7 - year follow - up study 显示文摘 | David AS Sara ER Robert CP | 2002 | Epilepsy &Behavior2002，3，： | 1 |
| 12 | Abnormal visual scan paths:a psychophysio- logical marker of delusions in schizophrenia 显示文摘 | Phillips ML David AS | 1998 | Schizophr Res1998，29，： | 1 |
| 13 | An evaluation of process and out comes from learning through reflective practice groups on a post -registration nursing course显示文摘 | Hazel PL David BL Dorothy AS | 2000 | J of Adv Nur2000，31，3： | 1 |
| 14 | Insight and psychosis显示文摘 | David AS | 1990 | Br J Psychiatry1990，156，： | 1 |
| 15 | Human papillomavirus- like particles mediate functional delivery of plasmid DNA to aotigen presenting cells in vivo显示文摘 | Christine MM David AS Young-Eun EL | 2007 | Vaccine2007，25，： | 1 |
| 16 | Tuberculosis of the musculoskeletal system显示文摘 | David AS Girish KS Ashok KB | 2005 | Techniques in Orthopaedics2005，20，2： | 1 |
| 17 | Tuberculosis of the Musculoskeletal System 显示文摘 | David AS Girish KS Ashok KB | 2005 | Techniques in Orthopaedics2005，20，2： | 1 |
| 18 | Retinoids in cancer therapy显示文摘 | Malcolm AS David RP Bruce DC | 1992 | J Clin Oncol1992，10，5： | 1 |
| 19 | Diffusion-weighted echo-planar MR imaging of CNS involvement in systemic lupuserythematosus显示文摘 | Toshil M David AS Yuji N | 2001 | Acad Radiol2001，8，8： | 1 |
| 20 | An evaluation of process and outcomes from learning through reflective practice groups on a post-registration nursing course显示文摘 | Hazel PL David BL Dorothy AS | 2000 | J of Adv Nur2000，31，3： | 1 |
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