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Pronuclear formation by ICSI using chemically activated ovine oocytes and zona pellucida bound sperm

Journal of Animal Science and Biotechnology 2017年第2期8卷 370-377页

作者： J.E.Hernández-Pichardo Y.Ducolomb S.Romo M.E.Kjelland R.fierro f.casillas M.Betancourt División de Ciencias Biológicas y de la Salud Departamento de Producción Agrícola y AnimalUniversidad Autónoma Metropolitana-XochimilcoCiudad de MexicoMexico Division de Ciencias Biologicas y de la Salud Departamento de Ciencias de la SaludUniversidad Autónoma Metropolitana-IztapalapaSan Rafael Atlixco 186CP 09340 Ciudad de MexicoMexico Departamento de Ciencias Pecuarias Facultad de EstudiosSuperiores CuautitlanUNAMCiudad de MexicoEstado de MexicoMexico Conservation Genetics&BiotechLLCValley CityNDUSA Division de Ciencias Biologicas y de la Salud Departamento de Biologia de la ReproduccionUniversidad Autonoma Metropolitana-lztapalapaCiudad de MexicoMexico Doctorado en Biologia Experimental Universidad Autonoma Metropolitana-lztapalapaCiudad de MexicoMexico

Background： In order to improve ICSI, appropiate sperm selection and oocyte activation is necessary. The objective of the present study was to determine the efficiency of fertilization using ICSI with chemically activated ovine oocytes and sperm selected by swim up（SU） or swim up ＋ zona pellucida（SU ＋ ZP） binding.Results： Experiment 1, 4–20 replicates with total 821 in vitro matured oocytes were chemically activated with ethanol, calcium ionophore or ionomycin, to determine oocyte activation（precense of one PN）. Treatments showed similar results（54, 47, 42 %, respectively） but statistically differents（P 0.05）.Conclusions： Chemical activation induces higher ovine oocyte activation than mechanical activation. Ethanol slightly displays higher oocyte activation than calcium ionophore and ionomicine. Sperm selection with SU ＋ ZP increased AR/A and AR/D rates in comparison with SU in fresh and frozen-thawed sperm. According to this, in terms of fertilization rates, chemical activation after ICSI increased oocyte PN formation compared to mechanical activation. Also, fresh sperm treated with SU and SU ＋ ZP were significantly different than frozen-thawed sperm,but between sperm treatments no significant differences were obtained.

关键词： Ethanol ICSI Oocyte activation Ovine Pronucleus Zona pellucida

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ACE Score Identifies HBeAg-negative Inactive Carriers at a Single-point Evaluation,Regardless of HBV Genotype

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Journal of Clinical and Translational Hepatology 2022年第6期10卷 1068-1076页

作者： Luisa Roade Mar Riveiro-Barciela Adriana Palom francisco Rodríguez-frías Marta Bes Ariadna Rando María Teresa Salcedo Rosario casillas Elena Vargas-Accarino David Tabernero Silvia Sauleda Rafael Esteban María Buti Liver Unit Internal Medicine DepartmentHospital Universitari Vall d’HebronVall d’Hebron Barcelona Hospital CampusBarcelonaSpain Universitat Autònoma de Barcelona(UAB) Department of MedicineBarcelonaSpain Universitat Au-tònoma de Barcelona(UAB) Department of Biochemistry and Molecular BiologyBarcelonaSpain Centro de Investigación Biomédica en Red de Enfermedades Hepáticas y Digestivas(CIBERehd) Instituto de Salud Carlos IIIMadridSpain Liver Pathology Lab Biochemistry and Microbiology Departments(Clinical Laboratories)Hospital Universitari Vall d’HebronVall d’Hebron Barcelona Hospital CampusBarcelonaSpain Vall d’Hebron Institut de Recerca(VHIR) Liver Diseases GroupBarcelonaSpain Transfusion Safety Laboratory Banc de Sang i TeixitsServei Catalàde la SalutBarcelonaSpain De-partment of Pathology Hospital Universitari Vall d’HebronUniversitat Autònoma de BarcelonaBarcelonaSpain

Background and Aims:Hepatitis B virus(HBV)biomark-ers have been used for a better categorization of patients,even though the lack of simple algorithms and the impact of genotypes limit their application.Our aim was to assess the usefulness of noninvasive markers for the identification of HBV inactive carriers(ICs)in a single-point evaluation and to design a predictive model for their identification.Meth-ods:This retrospective-prospective study included 343 consecutive HBeAg-negative individuals.Clinical,analytical,and virological data were collected,and a liver biopsy was performed if needed.Subjects were classified at the end of follow-up as ICs,chronic hepatitis B and gray zone.A pre-dictive model was constructed,and validated by 1000-boot-strap samples.Results:After 39 months of follow-up,298 subjects were ICs,36 were chronic hepatitis B CHB,and nine were gray zone.Eighty-nine(25.9%)individuals re-quired a liver biopsy.Baseline HBV DNA hazard ratio(HR)6.0,pf 0.925(95%CI:0.880-0.970,pfor identification of ICs.The AUROC for genotype D was 0.95,0.96 for A,0.90 for E,and 0.88 for H/f.An ACE score off 99.5%,and a score≤12 points had a diagnostic accuracy of 93.8%.Conclusions:Low baseline HBV DNA,HBcrAg,and liver stiffness were in-dependently associated with the IC phase.A score including those variables identified ICs at a single-point evaluation,and might be applied to implement less intensive follow-up strategies.

关键词： Hepatitis B virus Inactive carrier Liver stiffness HBV DNA Quan-titative HBsAg Core-related antigen.

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Evaluating physical changes of iron oxide nanoparticles due to surface modification with oleic acid

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Chinese Physics B 2020年第10期29卷 186-194页

作者： S Rosales N casillas A Topete O Cervantes G González J A Paz M E Cano Centro Universitario de Ciencias Exactas e Ingenienas Universidad de GuadalajaraBlvd.Marcelino Garcia Burragan 1421C.P.44430GuadalajaraJaliscoMexico Centro Universitario de la Cienegu Universidad de GuadalajaraAv.Universidad 1115C.P.47820OcotlanJaliscoMexico Centro Universitario de Ciencias de In Salud Universidad de Guadalajara.Sierra Mojada 950C.P.44340GuadalajaraJaliscoMexico

The physical characterization of a colloidal system of superficially modified magnetic nanoparticles (MNPs) is presented. The system consists of oleic acid-coated iron oxide nanoparticles (OAMNP) suspended in water. A structural analysis is carried out by using standard physical techniques to determine the diameter and shape of the MNPs and also the width of the coating shell. The colloidal stability and the polydispersity index of this ferrofluid are determined by using Zeta potential measurements. Additionally, the magnetic characterization is conducted by obtaining the DC magnetization loops, and the blocking temperatures are determined according to the ZfC–fC protocol. finally, the values of power absorption density P of the ferrofluid are estimated by using a magneto-calorimetric procedure in a wide range of magnetic field amplitude H and frequency f. The experimental results exhibit spherical-like shape of OAMNP with (20 ± 4) nm in diameter. Due to the use of coating process, the parameters of the magnetization loops and the blocking temperatures are significantly modified. Hence, while the uncoated MNPs show a blocking state of the magnetization, the OAMNP are superparamagnetic above room temperature (300 K). furthermore, the reached dependence P versus f and P versus H of the ferrofluid with coated MNPs are clearly fitted to linear and quadratic correlations, respectively, showing their accordance with the linear response theory.

关键词： nanoparticles ferrofluid magnetic hyperthermia functionalization

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Analysis of hepatitis B virus preS1 variability and prevalence of the rs2296651 polymorphism in a Spanish population

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World Journal of Gastroenterology 2018年第6期24卷 680-692页

作者： 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 Liver Pathology Unit Departments of Biochemistry and MicrobiologyHospital Universitari Vall d’HebronUniversitat Autònoma de BarcelonaBarcelona 08035Spain Liver Unit Liver Disease Laboratory-Viral HepatitisVall d’Hebron Institut Recerca-Hospital Universitari Vall d’HebronUniversitat Autònoma de BarcelonaBarcelona 08035Spain Centro de Investigación Biomédica en Red de Enfermedades Hepáticas y Digestivas(CIBERehd) Instituto de Salud Carlos IIIMadrid 28029Spain Roche Diagnostics SL Sant Cugat del Vallès 08174Spain Liver Unit Department of Internal MedicineHospital Universitari Vall d'HebronUniversitat Autònoma de BarcelonaBarcelona 08035Spain

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-interactin

关键词： Hepatitis B virus Hepatitis B virus preS1 region sodium-taurocholate co-transporting polypeptide NTCP-interacting domain virion morphogenesis domain SNP rs2296651 next-generation sequencing real-time PCR melting curves

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Effect of the sintering technique on the ferroelectric and d33 piezoelectric coefficients of Bi0.5(Na0.84K0.16)0.5TiO3 ceramic

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Journal of Advanced Ceramics 2019年第2期8卷 278-288页

作者： G.HERNANDEZ-CUEVAS J.R.LEYVA MENDOZA P.E.GARCíA-casillas C.A.RODRíGUEZ GONZáLEZ J.f.HERNANDEZ-PAZ G.HERRERA-PéREZ L.fUENTES-COBAS S.DíAZ DE LA TORRE O.RAYMOND-HERRERA H.CAMACHO-MONTES Instituto de Ingeniería y Tecnología Universidad Autónoma de Ciudad JuárezAv.Del Charro 450 Norte Cd.JuárezChihuahua32310México Centro de Investigación en Materiales Avanzados Miguel de Cervantes 120Chihuahua 31109ChihuahuaMéxico Centro de Investigación e Innovación Tecnológica Instituto Politécnico NacionalCerrada de Cecati s/nAzcapotzalcoSanta Catarina02250Ciudad de MéxicoMéxico Centro de Nanociencias y Nanotecnología Universidad Nacional Autónoma de MéxicoKm.107 Carretera Tijuana-EnsenadaAP.14Ensenada 22860Baja CaliforniaMéxico

In the search of lead-free piezoelectric materials,ceramic processing techniques offer potential tools to increase the piezoelectric and ferroelectric properties in addition to new chemical compositions.Powders of pure BNKT16(Bi0.5(Na0.84K0.16)0.5TiO3)phase were synthesized by sol–gel method with a low crystallization temperature(750℃).Ceramic samples were sintered by pressureless sintering(PLS),sinter-forging(Sf),and spark plasma sintering(SPS)techniques.Structural,morphological,and chemical characterizations were performed by XRD,Raman,EDS,and SEM.Sintered samples by PLS and Sf exhibit rod-like grains associated to bismuth volatility.The highest remanent polarization(11.05μC/cm2),coercive field(26.2 kV/mm),and piezoelectric coefficient(165 pC/N)were obtained for Sf sample.The piezoresponse force microscopy(PfM)analysis shows that the crystallites at the nanoscale exhibit piezoelectric phenomenon and the highest piezoelectric response is reported for PLS sample.The presence of the rhombohedral phase,the increase in grain and crystallite size,and the oriented rod-like inclusions favoring the crystallographic texture are facts that enhance the piezoelectric coefficient for BNKT16 piezoceramics.

关键词： lead-free piezoelectrics sol–gel pressureless sintering (PLS) sinter-forging (Sf) spark plasma sintering(SPS) piezoresponse

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Detection of hyper-conserved regions in hepatitis B virus X gene potentially useful for gene therapy

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World Journal of Gastroenterology 2018年第19期24卷 2095-2107页

作者： 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 Liver Pathology Unit Departments of Biochemistry and MicrobiologyHospital Universitari Vall d’HebronUniversitat Autònoma de BarcelonaBarcelona 08035Spain Liver Unit Liver Disease Laboratory-Viral HepatitisVall d’Hebron Institut Recerca-Hospital Universitari Vall d’HebronUniversitat Autònoma de BarcelonaBarcelona 08035Spain Centro de Investigación Biomédica en Red de Enfermedades Hepáticas y Digestivas(CIBERehd) Instituto de Salud Carlos IIIMadrid 28029Spain Roche Diagnostics SL Sant Cugat del Vallès 08174Spain Liver Unit Department of Internal MedicineHospital Universitari Vall d'HebronUniversitat Autònoma de BarcelonaBarcelona 08035Spain

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.

关键词： Hepatitis B virus Hepatitis B X gene Hepatitis B X protein Gene therapy Next-generation sequencing HBV conserved regions Small interference RNA

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