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Gene therapy is a promising approach for hereditary deafness. We recently showed that unilateral AAV1-hOTOF gene therapy with dual adeno-associated virus (AAV) serotype 1 carrying human OTOF transgene is safe and associated with functional improvements in patients with autosomal recessive deafness 9 (DFNB9). The protocol was subsequently amended and approved to allow bilateral gene therapy administration. Here we report an interim analysis of the single-arm trial investigating the safety and efficacy of binaural therapy in five pediatric patients with DFNB9. The primary endpoint was dose-limiting toxicity at 6 weeks, and the secondary endpoint included safety (adverse events) and efficacy (auditory function and speech perception). No dose-limiting toxicity or serious adverse event occurred. A total of 36 adverse events occurred. The most common adverse events were increased lymphocyte counts (6 out of 36) and increased cholesterol levels (6 out of 36). All patients had bilateral hearing restoration. The average auditory brainstem response threshold in the right (left) ear was >95 dB (>95 dB) in all patients at baseline, and the average auditory brainstem response threshold in the right (left) ear was restored to 58 dB (58 dB) in patient 1, 75 dB (85 dB) in patient 2, 55 dB (50 dB) in patient 3 at 26 weeks, and 75 dB (78 dB) in patient 4 and 63 dB (63 dB) in patient 5 at 13 weeks. The speech perception and the capability of sound source localization were restored in all five patients. These results provide preliminary insights on the safety and efficacy of binaural AAV gene therapy for hereditary deafness. The trial is ongoing with longer follow-up to confirm the safety and efficacy findings. Chinese Clinical Trial Registry registration: ChiCTR2200063181 .
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Dependovirus , Terapia Genética , Humanos , Terapia Genética/métodos , Niño , Masculino , Femenino , Dependovirus/genética , Preescolar , Sordera/genética , Sordera/terapia , Adolescente , Resultado del Tratamiento , Genes Recesivos , Vectores Genéticos/genética , Potenciales Evocados Auditivos del Tronco EncefálicoRESUMEN
Necrotic enteritis (NE) is a potentially fatal poultry disease that causes enormous economic losses in the poultry industry worldwide. The study aimed to evaluate the effects of dietary organic yeast-derived selenium (Se) on immune protection against experimental necrotic enteritis (NE) in commercial broilers. Chickens were fed basal diets supplemented with different Se levels (0.25, 0.50, and 1.00 Se mg/kg). To induce NE, Clostridium perfringens (C. perfringens) was orally administered at 14 days of age post hatch. The results showed that birds fed 0.25 Se mg/kg exhibited significantly increased body weight gain compared with the non-supplemented/infected birds. There were no significant differences in gut lesions between the Se-supplemented groups and the non-supplemented group. The antibody levels against α-toxin and NetB toxin increased with the increase between 0.25 Se mg/kg and 0.50 Se mg/kg. In the jejunal scrapings and spleen, the Se-supplementation groups up-regulated the transcripts for pro-inflammatory cytokines IL-1ß, IL-6, IL-8, iNOS, and LITAF and avian ß-defensin 6, 8, and 13 (AvBD6, 8 and 13). In conclusion, supplementation with organic yeast-derived Se alleviates the negative consequences and provides beneficial protection against experimental NE.
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Alimentación Animal , Pollos , Infecciones por Clostridium , Clostridium perfringens , Citocinas , Suplementos Dietéticos , Enteritis , Enfermedades de las Aves de Corral , Selenio , Animales , Enteritis/prevención & control , Enteritis/veterinaria , Enteritis/inmunología , Enteritis/microbiología , Selenio/farmacología , Selenio/administración & dosificación , Enfermedades de las Aves de Corral/prevención & control , Enfermedades de las Aves de Corral/inmunología , Clostridium perfringens/inmunología , Infecciones por Clostridium/prevención & control , Infecciones por Clostridium/veterinaria , Infecciones por Clostridium/inmunología , Citocinas/metabolismo , Toxinas Bacterianas/inmunología , Necrosis , beta-Defensinas/metabolismo , Yeyuno/efectos de los fármacos , Yeyuno/inmunología , Yeyuno/microbiología , Yeyuno/patología , Bazo/inmunología , Levaduras , Óxido Nítrico Sintasa de Tipo II/metabolismo , Interleucina-6/metabolismo , Interleucina-8/metabolismo , Interleucina-1beta/metabolismo , Anticuerpos Antibacterianos/sangreRESUMEN
OBJECTIVE: To investigate the audiological characteristics of vestibular schwannoma (VS) patients with normal pure-tone audiometry (PTA) results. STUDY DESIGN: A retrospective study. SETTING: Forty-two VS patients with normal PTA results from October 2016 to October 2022 were included. METHODS: Normal PTA was defined when the hearing threshold is ≤25 dB hearing loss (HL) in each test frequency and the PTA is ≤25 dB HL. Results of multiple audiological tests such as the auditory brainstem response (ABR), distortion product otoacoustic emission (DPOAE), multiple auditory steady-state responses threshold (ASSR), and speech discrimination score were retrospectively reviewed. Demographic data of these patients were also been collected. RESULTS: According to our results, the ABR and average ASSR threshold of the affected side were statistically significantly higher in VS patients with normal PTA. ABR waveforms on the affected side also showed more abnormalities. The DPOAE pass rates of the affected side were lower than the unaffected side while the amplitude and signal-to-noise ratio rate was also lower. In addition, we used magnetic resonance imaging 3-dimensional reconstruction images to measure the volume of tumors in these patients. We also found that higher ABR threshold means lager tumor size in patients with normal PTA. CONCLUSION: VS patients with normal PTA result cannot be assumed to have no impairment of hearing function. ABR, DPOAE, and ASSR results showed the characteristic changes in the affect ear. ABR threshold has the highest sensitivity for hearing abnormalities and is strong relative with tumor size in patients with normal PTA.
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Audiometría de Tonos Puros , Umbral Auditivo , Potenciales Evocados Auditivos del Tronco Encefálico , Neuroma Acústico , Humanos , Neuroma Acústico/complicaciones , Neuroma Acústico/fisiopatología , Femenino , Masculino , Estudios Retrospectivos , Persona de Mediana Edad , Adulto , Umbral Auditivo/fisiología , Potenciales Evocados Auditivos del Tronco Encefálico/fisiología , Anciano , Emisiones Otoacústicas Espontáneas/fisiologíaRESUMEN
BACKGROUND: Autosomal recessive deafness 9, caused by mutations of the OTOF gene, is characterised by congenital or prelingual, severe-to-complete, bilateral hearing loss. However, no pharmacological treatment is currently available for congenital deafness. In this Article, we report the safety and efficacy of gene therapy with an adeno-associated virus (AAV) serotype 1 carrying a human OTOF transgene (AAV1-hOTOF) as a treatment for children with autosomal recessive deafness 9. METHODS: This single-arm, single-centre trial enrolled children (aged 1-18 years) with severe-to-complete hearing loss and confirmed mutations in both alleles of OTOF, and without bilateral cochlear implants. A single injection of AAV1-hOTOF was administered into the cochlea through the round window. The primary endpoint was dose-limiting toxicity at 6 weeks after injection. Auditory function and speech were assessed by appropriate auditory perception evaluation tools. All analyses were done according to the intention-to-treat principle. This trial is registered with Chinese Clinical Trial Registry, ChiCTR2200063181, and is ongoing. FINDINGS: Between Oct 19, 2022, and June 9, 2023, we screened 425 participants for eligibility and enrolled six children for AAV1-hOTOF gene therapy (one received a dose of 9 × 1011 vector genomes [vg] and five received 1·5 × 1012 vg). All participants completed follow-up visits up to week 26. No dose-limiting toxicity or serious adverse events occurred. In total, 48 adverse events were observed; 46 (96%) were grade 1-2 and two (4%) were grade 3 (decreased neutrophil count in one participant). Five children had hearing recovery, shown by a 40-57 dB reduction in the average auditory brainstem response (ABR) thresholds at 0·5-4·0 kHz. In the participant who received the 9 × 1011 vg dose, the average ABR threshold was improved from greater than 95 dB at baseline to 68 dB at 4 weeks, 53 dB at 13 weeks, and 45 dB at 26 weeks. In those who received 1·5 × 1012 AAV1-hOTOF, the average ABR thresholds changed from greater than 95 dB at baseline to 48 dB, 38 dB, 40 dB, and 55 dB in four children with hearing recovery at 26 weeks. Speech perception was improved in participants who had hearing recovery. INTERPRETATION: AAV1-hOTOF gene therapy is safe and efficacious as a novel treatment for children with autosomal recessive deafness 9. FUNDING: National Natural Science Foundation of China, National Key R&D Program of China, Science and Technology Commission of Shanghai Municipality, and Shanghai Refreshgene Therapeutics.
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Dependovirus , Terapia Genética , Humanos , Terapia Genética/métodos , Dependovirus/genética , Niño , Masculino , Preescolar , Femenino , Adolescente , Lactante , Vectores Genéticos , Resultado del Tratamiento , Sordera/genética , Sordera/terapia , Mutación , Proteínas de la MembranaRESUMEN
To understand the epizootiologic characteristics of pathogens and opportunistic infections in one Beagle dog production colony and three research facilities, viruses and mycoplasma were detected in 1777 samples collected from Beagle dogs in China by polymerase chain reaction/reverse transcription polymerase chain reaction, and bacteria were isolated and identified by 16S rRNA sequence analysis. In addition, genotyping of the major circulating viruses was carried out by amplification of gene fragments and homology analysis. Canine coronavirus (CCoV), Escherichia coli, canine parvovirus (CPV), Bordetella bronchiseptica, Clostridium perfringens, Mycoplasma cynos, Klebsiella pneumoniae, Streptococcus canis, canine astrovirus (CaAstV), canine kobuvirus (CaKV), Pseudomonas aeruginosa, Proteus mirabilis, Macrococcus canis, Pasteurella canis, canine bocavirus (CBoV) and canine adenovirus (CAdV) were detected in the samples. Single, double, triple and quadruple infections accounted for 6.6%, 1.4%, 1.2% and 0.96% of samples, respectively. CCoV strains in 81 samples included three genotypes, CCoV-I, CCoV-IIa and CCoV-IIb, by analysis of S gene. The rate of single infection of CCoV-I, CCoV-IIa or CCoV-IIb was 19%, 38% or 7.4% respectively. The double and triple infection rates of CCoV were 32.8% and 2.5% respectively. All CPV strains in 36 samples belonged to CPV-2c. There were three amino acid differences in the Fiber protein of CAdV-positive sample QD2022, compared with the reference strain Toronto A26/61 and the vaccine strain YCA-18. These results suggest that CCoV and CPV are primary infectious agents, and that these two viruses were often identified in mixed infections, or coinfections alongside mycoplasma or other bacteria. These results will provide the basis for improvements in prevention and control of naturally occurring infectious diseases in Beagle dog production colonies and research facilities.
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Infecciones por Coronavirus , Coronavirus Canino , Enfermedades de los Perros , Parvovirus Canino , Perros , Animales , Infecciones por Coronavirus/epidemiología , Infecciones por Coronavirus/veterinaria , ARN Ribosómico 16S/genética , Enfermedades de los Perros/epidemiología , Reacción en Cadena de la Polimerasa , China/epidemiología , Coronavirus Canino/genética , Parvovirus Canino/genéticaRESUMEN
To detect the antibody against fowl adenovirus serotype 4 (FAdV-4) in clinical practice, the latex agglutination test (LAT) was developed by using the Fiber-2 protein of FAdV-4 as an antigen bound to sensitized latex microspheres. The concentration, time, and temperature of sensitization latex microspheres by the Fiber-2 protein were studied and optimized; the specificity, sensitivity, and repeatability of LAT were tested; and the method developed in the study was applied. The results showed that the optimum sensitization concentration of Fiber-2 protein was 0.8 mg/mL, the time was 120 min, and the temperature was 37 â. Except for antiserum against FAdV-4 and FAdV-10, LAT developed in the study could not agglutinate antisera against FAdV-1, FAdV-2, FAdV-3, FAdV-4, FAdV-5, FAdV-6, FAdV-8a, FAdV-8b, FAdV-11, Newcastle disease virus, infectious bronchitis virus, egg drop syndrome virus and Clostridium perfringens. Compared with the commercial FAdV-4 ELISA Kit, the titers in 21 clinical samples were low when tested by the developed LAT method, but there was no significant difference. The coefficients of variation among different batches and the same batch of latex-sensitized particles were between 0 % and 13.3 % and 0-8.7 %, respectively. The critical value of immune protective antibody against FAdV-4 was 25, and the titers in 40.9 % of clinical samples were higher than the immune critical point. The results showed that the Fiber-2-based LAT developed in the study has the characteristics of high specificity, sensitivity and repeatability, has the advantages of free equipment, long shelf life, and fast and easy operation, and is an effective and convenient method for serological diagnosis of FAdV-4 infection and evaluating the efficacy of vaccines.
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Infecciones por Adenoviridae , Aviadenovirus , Enfermedades de las Aves de Corral , Animales , Serogrupo , Infecciones por Adenoviridae/diagnóstico , Infecciones por Adenoviridae/veterinaria , Pruebas de Fijación de Látex , Anticuerpos Antivirales , Pollos , Adenoviridae , Enfermedades de las Aves de Corral/diagnósticoRESUMEN
Organic pollutants pose a serious threat to water environment, thus it is essential to develop high-performance adsorbent to remove them from wastewater. Herein, nitrogen-doped magnetic porous carbon (M-PLAC) with three-dimensional porous structure was synthesized from lignin to adsorb methylene blue (MB) and tetracycline (TC) in wastewater. The calculated equilibrium adsorption amount by M-PLAC for MB and TC was 645.52 and 1306.00 mg/g, respectively. The adsorption of MB and TC on M-PLAC conformed to the pseudo-second-order kinetic model. The removal of MB by M-PLAC showed fast and efficient characteristics and exhibited high selectivity for TC in a binary system. In addition, M-PLAC was suitable for a variety of complex water environments and had good regeneration performance, demonstrating potential advantages in practical wastewater treatment. The organic pollutant adsorption by M-PLAC was attributed to electrostatic interaction, hole filling effect, hydrogen bonding, and the π-π interaction.
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Contaminantes Ambientales , Contaminantes Químicos del Agua , Carbono , Lignina/química , Adsorción , Aguas Residuales , Porosidad , Nitrógeno , Tetraciclina/química , Antibacterianos , Agua/química , Fenómenos Magnéticos , Contaminantes Químicos del Agua/química , CinéticaRESUMEN
Amine-functionalized magnetic microspheres (LS-Fe-NMA) were successfully prepared from lignosulfonate by magnetization and amine modification to remove Cr(VI) from industrial wastewater. In addition to SEM, FTIR, XPS, magnetic hysteresis curves and other characterizations, the adsorption tests were performed in real Cr(VI)-containing industrial wastewater. The experimental influencing factors, such as adsorbent types, LS content in the composites, adsorption time, dosages of adsorbent and adsorption temperature were systematically explored. The results of adsorption models fitting proved that the adsorption conformed to the Langmuir model and the pseudo-second-order kinetic model. At 45 °C, the calculated equilibrium adsorption amount by LS-Fe-NMA in industrial wastewater was 642.11 mg/g. In addition, the adsorption process followed the principle of spontaneous endothermic reaction. It was found that electrostatic interaction, redox and complexation existed during the adsorption process. The excellent adsorption performance and reusability of LS-Fe-NMA for Cr(VI) demonstrates its practical application value in real Cr(VI)-containing industrial wastewater.
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Contaminantes Químicos del Agua , Purificación del Agua , Aguas Residuales , Microesferas , Contaminantes Químicos del Agua/análisis , Cromo/análisis , Adsorción , Purificación del Agua/métodos , Cinética , Fenómenos Magnéticos , Concentración de Iones de HidrógenoRESUMEN
Magnetic porous adsorbent materials are widely favored for their large specific surface area, good adsorption performance, and ease of separation. This work provided a magnetic biochar derived from furfural residue (M-FRAC) with excellent adsorption properties for various pollutants, including Congo red (CR), Tetracycline (TC), Bisphenol A (BPA), and Cr6+. The influence of experimental parameters, such as pollutant concentration, contact time, and pH, on the adsorption properties of M-FRAC was studied in detail. The adsorption process was highly dependent on pH and initial contaminant concentration. All pollutant adsorption was favorable under acidic conditions. The optimal pH of the CR, TC, and Cr6+ adsorption was 5, 4, and 2, respectively, while that of BPA was in the range of 2-5. The experimental equilibrium adsorption amount of CR, TC, BPA, and Cr6+ by M-FRAC was 110.89, 602.81, 157.76, and 265.31 mg/g, respectively. The adsorption processes of pollutants on M-FRAC were in accordance with the Langmuir isotherm model. The adsorption kinetics fitted the pseudo-second-order (PSO) kinetics model. In addition, M-FRAC could be readily separated from solution by applying an external magnetic field. Therefore, the M-FRAC has a good application prospect in practical industrial wastewater treatment.
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Contaminantes Ambientales , Contaminantes Químicos del Agua , Rojo Congo , Adsorción , Furaldehído , Tetraciclina , Antibacterianos , Carbón Orgánico/química , Fenómenos Magnéticos , Cinética , Contaminantes Químicos del Agua/análisis , Concentración de Iones de HidrógenoRESUMEN
BACKGROUND: Chronic subjective tinnitus poses significant challenges in clinical practice, and it is usually associated with hearing impairment, particularly with high-frequency sensorineural hearing loss (SNHL). Patients suffering from tinnitus with SNHL experience one of the most severe sensory disabilities, and this has devastating effects on their quality of life. Nowadays, mild to moderate SNHL can be managed with a properly fitted hearing aid (HA) that provides sound amplification, and several studies suggest that HAs may also benefit those with tinnitus. However, inadequate attention has been paid by medical personnel to the impact of HA use in residual hearing protection for patients with tinnitus and coexisting SNHL, and existing evidence is still at a preliminary stage. This study aims to identify and evaluate the efficacy of the use of HAs in both sound perception and residual hearing preservation among patients with tinnitus and coexisting SNHL. METHODS AND DESIGN: The present study is a prospective, single-center, outcome assessor and data analyst-blinded, randomized, controlled trial. Eligible participants will be recruited and randomly allocated into the HA intervention group and the waiting list control group at a ratio of 1:1. The primary outcome is to evaluate the severity of tinnitus using the Tinnitus Handicap Inventory as a continuous variable at 6 months from randomization. Secondary outcome measures include changes in hearing status and mental states. The trial will last 6 months, with follow-up visits at 3 months and 6 months. DISCUSSION: This will be the first randomized, controlled trial to identify and evaluate HAs' efficacy on residual hearing preservation among tinnitus patients with coexisting high-frequency SNHL in China. We are aiming for novelty and generalizability, and strengths of this study are that it will examine the effectiveness of HA in patients with tinnitus and hearing impairment and will further explore the residual hearing protection provided by HA treatment in the tinnitus group. TRIAL REGISTRATION: ClinicalTrials.gov NCT05343026. Registered on April 25, 2022.
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Sordera , Audífonos , Pérdida Auditiva Sensorineural , Pérdida Auditiva , Acúfeno , Humanos , Acúfeno/diagnóstico , Acúfeno/terapia , Acúfeno/complicaciones , Calidad de Vida , Estudios Prospectivos , Resultado del Tratamiento , Audición , Pérdida Auditiva Sensorineural/diagnóstico , Pérdida Auditiva Sensorineural/terapia , Percepción , Ensayos Clínicos Controlados Aleatorios como AsuntoRESUMEN
Five novel chicken astrovirus (CAstV) strains, designated ZDF, MHC, WSC, WSW and MHW, were successfully isolated from chickens with gout, and were subjected to full genome sequencing characterization and tested for their pathogenic effects in specific pathogen-free (SPF) chicken embryos and chickens. The complete genomes of the five isolated strains were approximately 7436â nt to 7511â nt in length. Phylogenetic analysis revealed that strains ZDF and MHC were clustered in a clade with strains isolated in China and that the others were clustered with strains from other countries. Based on the amino acids of ORF2, strains MHW and WSW belonged to subgroup Ai, strain WSC belonged to Bii, and strains ZDF and MHC belonged to Bi. The pathogenicity of strains MHW, MHC and WSC, all belonging to different subgroups was studied. The results showed that the mortality of the chicken embryos was 100% when infected with any strain at a dose of more than 103 TCID50, 35% in SPF chickens infected with strain WSC, 25% with MHC and 15% with MHW. The body weights of chickens and embryos infected with 0.2 ml 10 TCID50 were significantly reduced after hatching. SPF chickens infected with any of the strains had similar lesions characterized by urate deposits on the epicardium and kidney, and necrotic spots on the liver. This study identified the three types of genotypic CAstV prevalent in China, with high mortality in embryonated chicken eggs and leading to white chick syndrome, retarded growth and visceral gout in infected chicks.
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Infecciones por Astroviridae , Avastrovirus , Gota , Enfermedades de las Aves de Corral , Animales , Infecciones por Astroviridae/veterinaria , Embrión de Pollo , Pollos , China/epidemiología , Variación Genética , Gota/veterinaria , Filogenia , VirulenciaRESUMEN
Age-related changes in human vaginal microbiota composition have been reported, and such changes might be influenced by humidity, external stimuli, hormone levels, drug to use, and other factors. However, there is no report about the vaginal microbiota composition of female beagles at different ages. To investigate the effects of aging on the vaginal microbiota independent of other effects, we analyzed the vaginal microbiomes of 23 beagles at a wide range of ages from 1 year to 7 years old (except the 3rd year), 1-2 y were categorized into youth stage (YS), 4-5 y were categorized into middle stage (MS), and 6-7 y were categorized into elderly stage (ES) based on age. Samples were collected by scraping the vaginal mucosa of YS (n = 8), MS (n = 5) and ES (n = 10), and analyzed by 16S-rRNA gene high-throughput-sequencing. The diversity of the vaginal microbiome in female beagles was found to continuously change with age. We also found associations between age and specific members and functions of the vaginal microbiome. The metabolism of terpenoids and polyketide and the cell motility are significantly enhanced with age. Our results suggest that the proportion of Tenericutes might be a biomarker which could distinguish between YS and others.
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Microbiota , Animales , Perros , Femenino , Secuenciación de Nucleótidos de Alto Rendimiento/veterinaria , Humanos , ARN Ribosómico 16S/genética , VaginaRESUMEN
Fowl adenovirus (FAdV) infections in chickens have resulted in global economic losses in the poultry industry. Infectious bursal disease virus (IBDV) and chicken anemia virus (CAV) infections lead to immunosuppression in chickens, and concomitant co- infection with FAdV usually produces severe and lethal infections. These co-infections are common occurrences on chicken farms and affect large number of chickens. Thus, a rapid, sensitive and specific diagnostic test for these viruses becomes a prerequisite to effective control and isolation measures. We developed a triplex nanoparticle-assisted PCR (nano-PCR) assay that can simultaneously detect these 3 viruses in a single assay tube using PCR primers directed at respective specific genes of each virus. The assay was specific for FAdVs, CAV and IBDV, and it did not amplify Newcastle disease virus, infectious bronchitis virus, egg drop syndrome virus or Marek's disease virus. The minimum detection limit was 27.2 femtogram (fg) for all three viruses and was 1000-fold more sensitive than multiplex PCR using identical primers. Screening of 69 clinical samples from 40 to 50 days old chickens with obvious lesions in liver using the nano-PCR compared with a multiplex PCR yielded identical results. Of the 69 samples, 13 were detected positive including 4 for FAdV, 4 for IBDV and 6 for CAV single virus infections, respectively, as well as 5 for FAdV/CAV, 2 for FAdV/IBDV and 3 for IBDV/CAV co-infections. The triple nano-PCR assay developed in our laboratory is a sensitive, specific and simple method that can be used for detection of FAdV, CAV and IBDV as single or mixed infections.
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Aviadenovirus , Virus de la Anemia del Pollo , Virus de la Enfermedad Infecciosa de la Bolsa , Nanopartículas , Enfermedades de las Aves de Corral , Animales , Aviadenovirus/genética , Virus de la Anemia del Pollo/genética , Pollos , Virus de la Enfermedad Infecciosa de la Bolsa/genética , Reacción en Cadena de la Polimerasa Multiplex , Enfermedades de las Aves de Corral/diagnósticoRESUMEN
OBJECTIVE: We sought to determine the biocompatibility of electrospun regenerated silk fibroin (RSF) mats with inner ear progenitors, especially their effect on the differentiation of inner ear progenitors into hair cells. METHODS: Neonatal mouse cochleae (n = 20) were collected and digested and allowed to form spheres over several days. Cells digested from the spheres were then seeded onto aligned or random RSF mats, with laminin-coated coverslips serving as controls. The inner ear progenitor cell mortality was examined by TUNEL labeling, and the adhesion of cells to the RSF mats or coverslip was determined by scanning electron microscopy. Finally, the number of hair cells that differentiated from inner ear progenitors was determined by Myosin7a expression. Unpaired Student's t-tests and one-way ANOVA followed by a Dunnett's multiple comparisons test were used in this study (p < 0.05). RESULTS: After 5 days of culture, the inner ear progenitors had good adhesion to both the aligned and random RSF mats and there was no significant difference in TUNEL+ cells between the mats compared to the coverslip (p > 0.05). After 7 days of in vitro differentiation culture, the percentage of differentiated hair cells on the control, aligned, and random RSF mats was 2.5 ± 0.5%, 2.7 ± 0.4%, and 2.4 ± 0.2%, respectively, and there was no significant difference between Myosin7a+ cells on either RSF mat compared to controls (p > 0.05). CONCLUSION: The aligned and random RSF mats had excellent biocompatibility with inner ear progenitors and helped the inner ear progenitors maintain their stemness. Our results thus indicate that RSF mats represent a useful scaffold for the development of new strategies for inner ear tissue engineering research.
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Oído Interno , Fibroínas , Animales , Materiales Biocompatibles , Humanos , Ratones , Seda , Adherencias Tisulares , Ingeniería de TejidosRESUMEN
Monolepta hieroglyphica (Motschulsky) (Coleoptera: Chrysomelidae) is an important agricultural insect pest. In this study, the complete mitochondrial genome of M. hieroglyphica was sequenced using Illumina HiSeq X Ten. The mitogenome was 16,213 bp long and comprised 13 protein-coding genes (PCGs), two ribosomal RNA genes (rRNAs), 22 transfer RNA genes (tRNAs) and a putative control region (CR). The nucleotide composition of the M. hieroglyphica mitochondrial genome was significantly biased (A, G, C and T accounted for 41.04%, 8.01%, 11.76% and 39.18%, respectively) with 80.23% A + T content. Two rRNAs were located between tRNA-Leu and the CR, separated by tRNA-Val. The CR, located between 12 s rRNA and tRNA-Ile, was 1661 bp long. The length of the 22 tRNAs ranged from 61 to 71 bp. Phylogenetic analyses of 29 Chrysomelidae-Galerucinae species based on 13 mitochondrial protein-coding genes reconstructed using Bayesian 3.2.0 showed that the M. hieroglyphica mitogenome was clustered with the existing three different species of the Monolepta genus mitogenomes in a monophyletic manner. The M. hieroglyphica mitogenome provides an important data resource for further studies and contributes to our understanding of the phylogeny of this group.
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Monolepta hieroglyphica (Motschulsky) (Coleoptera: Chrysomelidae) is an important agricultural insect pest. In this study, the complete mitochondrial genome of M. hieroglyphica (GenBank accession number MW732714) was sequenced using Illumina HiSeq X Ten. The mitogenome was 16,213 bp long and comprised 13 protein-coding genes (PCGs), two ribosomal RNA genes (rRNAs), 22 transfer RNA genes (tRNAs) and a putative control region (CR). The nucleotide composition of the M. hieroglyphica mitochondrial genome was significantly biased (A, G, C and T accounted for 41.04%, 8.01%, 11.76% and 39.18%, respectively) with 80.23% A + T content. Two rRNAs were located between tRNA-Leu and the CR, separated by tRNA-Val. The CR, located between 12 s rRNA and tRNA-Ile, was 1,661 bp long. The length of the 22 tRNAs ranged from 61 to 71 bp. Phylogenetic analyses of 29 Chrysomelidae-Galerucinae species based on 13 mitochondrial protein-coding genes reconstructed using Bayesian 3.2.0 showed that the M. hieroglyphica mitogenome was clustered with the existing three different species of the Monolepta genus mitogenomes in a monophyletic manner. The M. hieroglyphica mitogenome provides an important data resource for further studies and contributes to our understanding of the phylogeny of this group.
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H9N2 Avian influenza virus (AIV) is regarded as a principal donor of viral genes through reassortment to co-circulating influenza viruses that can result in zoonotic reassortants. Whether H9N2 virus can maintain sustained evolutionary impact on such reassortants is unclear. Since 2013, avian H7N9 virus had caused five sequential human epidemics in China; the fifth wave in 2016-2017 was by far the largest but the mechanistic explanation behind the scale of infection is not clear. Here, we found that, just prior to the fifth H7N9 virus epidemic, H9N2 viruses had phylogenetically mutated into new sub-clades, changed antigenicity and increased its prevalence in chickens vaccinated with existing H9N2 vaccines. In turn, the new H9N2 virus sub-clades of PB2 and PA genes, housing mammalian adaptive mutations, were reassorted into co-circulating H7N9 virus to create a novel dominant H7N9 virus genotype that was responsible for the fifth H7N9 virus epidemic. H9N2-derived PB2 and PA genes in H7N9 virus conferred enhanced polymerase activity in human cells at 33°C and 37°C, and increased viral replication in the upper and lower respiratory tracts of infected mice which could account for the sharp increase in human cases of H7N9 virus infection in the 2016-2017 epidemic. The role of H9N2 virus in the continual mutation of H7N9 virus highlights the public health significance of H9N2 virus in the generation of variant reassortants of increasing zoonotic potential.IMPORTANCEAvian H9N2 influenza virus, although primarily restricted to chicken populations, is a major threat to human public health by acting as a donor of variant viral genes through reassortment to co-circulating influenza viruses. We established that the high prevalence of evolving H9N2 virus in vaccinated flocks played a key role, as donor of new sub-clade PB2 and PA genes in the generation of a dominant H7N9 virus genotype (G72) with enhanced infectivity in humans during the 2016-2017 N7N9 virus epidemic. Our findings emphasize that the ongoing evolution of prevalent H9N2 virus in chickens is an important source, via reassortment, of mammalian adaptive genes for other influenza virus subtypes. Thus, close monitoring of prevalence and variants of H9N2 virus in chicken flocks is necessary in the detection of zoonotic mutations.
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Jiang, Tao, Liping Zhao, Yanbo Yin, Huiqian Yu, and Qingzhong Li. Evaluation of middle ear function by tympanometry and the influence of lower barometric pressure at high altitude. High Alt Med Biol. 22:70-76, 2021. Objective: To evaluate middle ear function in residents of high-altitude areas in comparison to sea-level participants. Design: Prospective observational study. Setting: All experiments were performed by experienced audiologists with a calibrated tympanometry machine. Participants: Young adults between the age of 17 and 23 were recruited. Seventy-five participants from Shanghai (altitude 4 m) and 133 participants from the Shigatse area (altitude 4,040 m) were recruited. Any participant with any otological disorder was excluded. Main Outcome Measure: Four indexes of the tympanogram were evaluated in the two groups from different altitudes. Results: Our results showed that the peak of static compliance for the participants in Shigatse was smaller, but the absolute compliance of tympanic membrane remained the same. Similarly, the ear canal volume (ECV) from tympanometry was also affected by the elevated altitude in Tibet. In addition, the tympanometric peak pressure was decreased in high-altitude residents, which suggests a slightly declined function of the Eustachian tube at lower barometric pressure. However, no difference was found in the tympanometric width (TW). Conclusion: Our results indicate that tympanograms were affected by decreased atmospheric pressure at high altitude. Therefore, other than pressure-related indexes, TW is better for evaluating the function of the middle ear in high-altitude regions.
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Pruebas de Impedancia Acústica , Altitud , Presión Atmosférica , China , Oído Medio , Humanos , TibetRESUMEN
Ostrich diseases characterized by paralysis have been breaking out in broad areas of China since 2015, causing major damage to the ostrich breeding industry in China. This report describes a parvovirus detected in ostriches from four different regions. The entire genomes of four parvovirus strains were sequenced following amplification by PCR, and we conducted comprehensive analysis of the ostrich parvovirus genome. Results showed that the length genomes of the parvovirus contained two open reading frames. Ostrich parvovirus (OsPV) is a branch of goose parvovirus (GPV). Genetic distance analysis revealed a close relationship between the parvovirus and goose parvovirus strains from China, with the closest being the 2016 goose parvovirus RC16 strain from Chongqing. This is the first report of a parvovirus in ostriches. However, whether OsPV is the pathogen of ostrich paralysis remains uncertain. This study contributes new information about the evolution and epidemiology of parvovirus in China, which provides a new way for the study of paralysis in ostriches.
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Evolución Molecular , Genoma Viral , Infecciones por Parvoviridae/virología , Parvovirus/fisiología , Struthioniformes/virología , Animales , Secuencia de Bases , Pruebas Genéticas , Genómica/métodos , Infecciones por Parvoviridae/diagnóstico , Filogenia , Reacción en Cadena de la PolimerasaRESUMEN
Though blastocyst production in vitro has been successful in several animal species, a culture system to produce viable and normal canine blastocysts in vitro remains to be established. In this study, we report the development of an in vitro culture system for canine preimplantation embryos produced via parthenogenetic activation (PA) and somatic cell nucleus transfer (SCNT). Our results show that the medium developed by us, named "Qingdao Agricultural University's (QAU)-4 medium", successfully breaks the developmental arrest observed at the eight-cell stage in canine embryos grown in other culture systems. The blastocysts produced in QAU-4 displayed normal blastocyst structures, including a clear inner cell mass and blastocyst cavity. We also found that blastocyst formation in PA embryos cultured in QAU-4 medium was quite high, though this was not so in the case of SCNT embryos. However, supplementation of QAU-4 medium with 100â¯nM of scriptaid caused a sharp increase in blastocyst formation in SCNT embryos. After culture, hatched blastocysts were also observed to successfully adhere to collagen-coated dishes, where further growth and differentiation occurred. To our knowledge, this is the first in vitro canine preimplantation embryo culture system that can successfully produce canine blastocysts.