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The electrocatalytic carbon dioxide reduction (CO2RR) is one of the emerging technologies that can effectively transform carbon dioxide (CO2) into valuable products. Electrocatalysts deriving from green synthesis methods will significantly help to establish a new green carbon cycle. Herein, a green electrodeposition method without additional reducing agents was used to synthesize Cu-Ag bimetallic catalysts, and it is shown that the combination of Cu and Ag obviously affects the morphology of the Cu-Ag catalysts, resulting in the formation of elaborate tree-like Cu-Ag clusters. An as-deposited Cu-Ag/carbon fiber (Cu-Ag/CF) catalyst exhibits high activity, selectivity and stability toward the CO2RR; in particular, the elaborate dendritic Cu-Ag/CF can efficiently reduce CO2 to syngas with high selectivity (Faradaic efficiency (FE) > 95%) at a low onset potential (-0.5 V). This work provides a rational strategy to overcome the significantly different reaction capacities during the reduction of Ag+ and Cu2+, leading to the formation of a controlled morphology of Cu-Ag, which is favourable for the design and development of highly efficient Cu or Ag catalysts via green methods for electrocatalyzing the CO2RR.
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In the precision medicine of lung adenocarcinoma, the identification and prediction of tumor phenotypes for specific biomolecular events are still not studied in depth. Various earlier researches sheds light on the close correlation between genetic expression signatures and DNA copy number variations (CNVs), for which analysis of CNVs provides valuable information about molecular and phenotypic changes in tumorigenesis. In this study, we propose a comprehensive analysis combining genome-wide association analysis and an Elastic Net Regression predictive model, focus on predicting the levels of many gene expression signatures in lung adenocarcinoma, based upon DNA copy number features alone. Additionally, we predicted many other key phenotypes, including clinical features (pathological stage), gene mutations, and protein expressions. These Elastic Net prediction methods can also be applied to other gene sets, thereby facilitating their use as biomarkers in monitoring therapy.
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The Cu-In2O3/C nanocomposite was prepared by a simple solid-phase reduction method. The introduction of In2O3 into Cu/C to form the Cu-In2O3/C nanocomposite evidently enhances the electrocatalytic activity for the selective reduction of CO2 to CO. Specifically, the Cu-In2O3/C nanocomposite exhibits higher Faraday efficiency (FE = 86.7%) at -0.48 V vs. the reversible hydrogen electrode (RHE) in the electrocatalytic reduction of CO2 to CO and larger current densities (55 mA cm-2) under a low overpotential (-1.08 V vs. RHE). These indicate its superior performance over many of the reported Cu-based catalysts [1-4]. It was also found that by rationally adjusting the applied potential, tunable syngas can be formed, which can be used to synthesize formic acid, methyl ether, methanol, synthetic fuels, or other bulk chemicals through appropriate industrial processes. Furthermore, the Cu-In2O3/C nanocomposite maintains good stability in the electrocatalytic reduction of CO2. This work demonstrates a novel strategy to convert CO2 into desired products with high energy efficiency and large current density under low overpotential by the rational designing of non-precious metal catalysts.
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OBJECTIVE: Amyotrophic lateral sclerosis (ALS) is an adult-onset fatal neurodegenerative disease which lacks identified biological markers. A label-free plasma surface-enhanced Raman spectroscopy (SERS) method was developed to explore a simple and noninvasive test for ALS. METHODS: ALS patients were enrolled serially and plasma samples were collected at the time of diagnosis prior to the start of ALS treatment. SERS spectra were recorded using a Renishaw micro-Raman system. RESULTS: To exclude the interference by varying disease severity, we enrolled three groups of ALS patients, including ALS-1 (n = 60; ALSFRS-R ≥ 42 and time interval ≤ 12 months), ALS-2 (n = 61; ALSFRS-R < 42 and time interval ≤ 12 months), and ALS-3 (n = 61; ALSFRS-R ≥ 38 and time interval> 12 months). The SERS spectra were analyzed using principal component analysis (PCA), which showed that ALS-1, ALS-2, ALS-3, and control groups were separated significantly. Then, decision tree (DT) models and receiver operating characteristic curves were employed and identified that bands at 722 and 739 cm-1 , and ratios of 635-722 cm-1 and 635-739 cm-1 were able to distinguish ALS from controls significantly. Finally, we highlighted six metabolism pathways correlated with ALS, including phenylalanine-tyrosine-tryptophan biosynthesis, aminoacyl-tRNA biosynthesis, phenylalanine metabolism, pantothenate and CoA biosynthesis, porphyrin and chlorophyll metabolism, and pyrimidine metabolism. INTERPRETATION: Plasma SERS could be a promising tool for the detection of ALS. The bands at 722 and 739 cm-1 , and the ratios of 635-722 cm-1 and 635-739 cm-1 could serve as potential indicator for ALS.
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Esclerose Lateral Amiotrófica/sangue , Esclerose Lateral Amiotrófica/diagnóstico , Biomarcadores/análise , Doenças Neurodegenerativas/diagnóstico , Progressão da Doença , Feminino , Humanos , Pessoa de Meia-Idade , Doenças Neurodegenerativas/metabolismo , Plasma/metabolismo , Curva ROC , Índice de Gravidade de DoençaRESUMO
The original version of this article unfortunately contained mistakes in the affiliation section.
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An aqueous solution synthesis method was used to synthesize a Pt-Ni/GNs composite containing trace amounts of Ni species by the aid of self-etching or acid-etching process. Its component structure and morphology were characterized by X-ray diffraction (XRD), Transmission electron microscope (TEM), X-ray photoelectron spectroscopy (XPS), Inductively Coupled Plasma-Atomic Emission Spectroscopy(ICP-AES) and Raman spectra, etc. The obvious cracked Pt-Ni nanoclusters can be found in Pt-Ni (trace)/GNs which contains only trace Ni species. Electrochemical experiments indicate that Pt-Ni (trace)/GNs exhibits bi-functional electrocatalytic performance for MOR and ORR with the mass activity of 1009.98â¯mAâ¯mg-1 and 157.7â¯mAâ¯mg-1, respectively, which is superior to commercial Pt-Ru/C-JM. It is proven that the trace Ni species contribute to the enhanced electrocatalytic performance of the Pt-Ni/GNs composite.
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Autosomal recessive optic neuropathies (IONs) are extremely rare disorders affecting retinal ganglion cells and the nervous system. RTN4IP1 has recently been identified as the third known gene associated with the autosomal recessive ION optic atrophy 10 (OPA10). Patients with RTN4IP1 mutations show early-onset optic neuropathy that can be followed by additional neurological symptoms such as seizures, ataxia, mental retardation, or even severe encephalopathy. Here, we report two siblings from a Chinese family who presented with early-onset optic neuropathy, epilepsy, and mild intellectual disability. Using whole exome sequencing combined with Sanger sequencing, we identified novel compound heterozygous RTN4IP1 mutations (c.646G > A, p.G216R and c.1162C > T, p.R388X) which both co-segregated with the disease phenotype and were predicted to be disease-causing by prediction software. An in vitro functional study in urine cells obtained from one of the patients revealed low expression of the RTN4IP1 protein. Our results identify novel compound heterozygous mutations in RTN4IP1 which are associated with OPA10, highlighting the frequency of RTN4IP1 mutations in human autosomal recessive IONs. To our knowledge, this is the first report of RTN4IP1 carriers from China.
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Proteínas de Transporte/genética , Proteínas Mitocondriais/genética , Atrofia Óptica Hereditária de Leber/genética , Proteínas de Transporte/metabolismo , Criança , Feminino , Heterozigoto , Humanos , Proteínas Mitocondriais/metabolismo , Mutação , Atrofia Óptica Hereditária de Leber/patologia , Sequenciamento do ExomaRESUMO
Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease with a wide range of survival times. We aimed to explore prognostic factors related to short survival based on clinical features and plasma metabolic signatures using surface-enhanced Raman spectroscopy (SERS). One hundred and thirty-eight sporadic ALS cases were enrolled serially, including 62 for the short-duration group (≤3 years) and 76 for the long-duration group (>3 years). Multivariate analysis showed that an older age of onset (>60 years; odds ratio [OR] = 3.98, 95% CI: 1.09-14.53), lower body mass index (BMI) (<18.5; OR = 6.80, 95% CI: 1.36-33.92), and lower ALSFRS-R score (<35; OR = 6.03, 95% CI: 1.42-25.63) were associated with higher odds of tracheotomy or death, while a higher uric acid (UA) level showed a protective effect (>356.36 µmol/L; OR = 0.19, 95% CI: 0.05-0.73). SERS analysis showed significant differences between the two groups, and pathway analysis highlighted five main metabolic pathways, including metabolisms of glutathione, pyrimidine, phenylalanine, galactose, and phenylalanine-tyrosine-tryptophan biosynthesis. In conclusion, age of onset, BMI, ALSFRS-R score and UA, together with dysregulation of glucose, amino acid, nucleic acid, and antioxidant metabolism contributed to disease progression, and are therefore potential therapeutic targets for ALS.
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Esclerose Lateral Amiotrófica/sangue , Esclerose Lateral Amiotrófica/diagnóstico , Análise Espectral Raman , Feminino , Humanos , Processamento de Imagem Assistida por Computador , Estimativa de Kaplan-Meier , Masculino , Pessoa de Meia-Idade , PrognósticoRESUMO
Primary familial brain calcification (PFBC) is a rare neurological disorder. Mutations in five genes (SLC20A2, PDGFRB, PDGFB, XPR1, and MYORG) have been linked to PFBC. Here, we used SYBR green-based real-time quantitative polymerase chain reaction (PCR) assay and denaturing high-performance liquid chromatography analysis to detect copy number variants (CNVs) in 20 unrelated patients with PFBC, negatively sequenced for the five known genes. We identified three deletions in SLC20A2, including a large de novo full gene deletion and two exonic deletions confined to exon 2 and exon 6, respectively. Subsequent linked-read whole-genome sequencing of the patient with the large deletion showed a 1.7 Mb heterozygous deletion which removed the entire coding regions of SLC20A2 as well as 21 other genes. In the family with a deletion of exon 6, a missense variant of uncertain significance (SLC20A2: p.E267Q) also co-segregated with the disease. Functional assay showed the deletion could result in significantly impaired phosphate transport, whereas the p.E267Q variant did not. Our results confirm that deletion in SLC20A2 is a causal mechanism for PFBC and highlight the importance of functional study for classifying a rare missense variant as (likely) pathogenic.
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Doenças dos Gânglios da Base/diagnóstico , Doenças dos Gânglios da Base/genética , Calcinose/diagnóstico , Calcinose/genética , Estudos de Associação Genética , Predisposição Genética para Doença , Doenças Neurodegenerativas/diagnóstico , Doenças Neurodegenerativas/genética , Deleção de Sequência , Proteínas Cotransportadoras de Sódio-Fosfato Tipo III/genética , Adolescente , Adulto , Idoso , Alelos , Criança , Feminino , Genótipo , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Masculino , Repetições de Microssatélites , Pessoa de Meia-Idade , Linhagem , Fenótipo , Análise de Sequência de DNA , Receptor do Retrovírus Politrópico e Xenotrópico , Adulto JovemRESUMO
Primary familial brain calcification (PFBC) is a rare neurodegenerative disorder with four causative genes (SLC20A2, PDGFRB, PDGFB, and XPR1) that have been identified. Here, we aim to describe the mutational spectrum of four causative genes in a series of 226 unrelated Chinese PFBC patients. Mutations in four causative genes were detected in 16.8% (38/226) of PFBC patients. SLC20A2 mutations accounted for 14.2% (32/226) of all patients. Mutations in the other three genes were relatively rare, accounting for 0.9% (2/226) of all patients, respectively. Clinically, 44.8% of genetically confirmed patients (probands and relatives) were considered symptomatic. The most frequent symptoms were chronic headache, followed by movement disorders and vertigo. Moreover, the total calcification score was significantly higher in the symptomatic group compared to the asymptomatic group. Functionally, we observed impaired phosphate transport induced by seven novel missense mutations in SLC20A2 and two novel mutations in XPR1. The mutation p.D164Y in XPR1 might result in low protein expression through an enhanced proteasome pathway. In conclusion, our study further confirms that mutations in SLC20A2 are the major cause of PFBC and provides additional evidence for the crucial roles of phosphate transport impairment in the pathogenies of PFBC.
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Encefalopatias/genética , Calcinose/genética , Predisposição Genética para Doença , Mutação , Doenças Neurodegenerativas/genética , Adulto , Idoso , Alelos , Transporte Biológico , Biomarcadores , Encefalopatias/diagnóstico , Encefalopatias/metabolismo , Calcinose/diagnóstico , Calcinose/metabolismo , Linhagem Celular Tumoral , China , Feminino , Genes sis , Genótipo , Humanos , Masculino , Pessoa de Meia-Idade , Doenças Neurodegenerativas/diagnóstico , Doenças Neurodegenerativas/metabolismo , Neuroimagem , Fenótipo , Receptor beta de Fator de Crescimento Derivado de Plaquetas/genética , Receptores Acoplados a Proteínas G/genética , Receptores Virais/genética , Proteínas Cotransportadoras de Sódio-Fosfato Tipo III/genética , Tomografia Computadorizada por Raios X , Receptor do Retrovírus Politrópico e XenotrópicoRESUMO
Primary familial brain calcification (PFBC) is a genetically heterogeneous disorder characterized by bilateral calcifications in the basal ganglia and other brain regions. The genetic basis of this disorder remains unknown in a significant portion of familial cases. Here, we reported a recessive causal gene, MYORG, for PFBC. Compound heterozygous or homozygous mutations of MYORG co-segregated completely with PFBC in six families, with logarithm of odds (LOD) score of 4.91 at the zero recombination fraction. In mice, Myorg mRNA was expressed specifically in S100ß-positive astrocytes, and knockout of Myorg induced the formation of brain calcification at 9 months of age. Our findings provide strong evidence that loss-of-function mutations of MYORG cause brain calcification in humans and mice.
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Astrócitos/metabolismo , Encefalopatias/genética , Calcinose/genética , Glicosídeo Hidrolases/genética , Mutação com Perda de Função , RNA Mensageiro/metabolismo , Adulto , Idoso , Alelos , Animais , Estudos de Casos e Controles , Feminino , Humanos , Masculino , Camundongos , Camundongos Knockout , Pessoa de Meia-Idade , Mutação , LinhagemRESUMO
During an analysis of the virome of bats from Myanmar, a large number of reads were annotated to orthohepadnaviruses. We present the full genome sequence and a morphological analysis of an orthohepadnavirus circulating in bats. This virus is substantially different from currently known members of the genus Orthohepadnavirus and represents a new species.
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Quirópteros/virologia , Genoma Viral , Hepatite Viral Animal/epidemiologia , Orthohepadnavirus/genética , RNA Viral/genética , Animais , Hepatite Viral Animal/virologia , Mianmar/epidemiologia , Orthohepadnavirus/classificação , Orthohepadnavirus/isolamento & purificação , Filogenia , Reação em Cadeia da Polimerase , Prevalência , RNA Viral/classificação , RNA Viral/isolamento & purificação , Análise de Sequência de DNARESUMO
To develop a specific, rapid, and convenient immunochromatography assay (ICA) to detect the rabies antibody in clinical sample from immuned dogs by rabies vaccine. Colloidal gold particles labeled with purified rabies virus (CVS11) were used as the detector reagent. The staphylococcal protein A (SPA) and pured rabbit anti-rabies virus IgG were blotted on the test and control regions of nitrocellulose membrane. Then the strip was assembled with sample pad, absorbing pad, and dorsal shield. The assay samples (261 dog's serum) were collected from Wildlife Rabies Disease Diagnostic Laboratories of Ministry of Agriculture in China, Institute of Military Veterinary, Academy of Military Medical Sciences and other six provinces, including rabies virus positive and negative serum. The performance of the strip was compared to fluorescent antibody virus neutralization test. The neutralizing antibody titer could be detected above 0.5 IU. The strip did not change of performance when stored at room temperature for 12 months. It may offer reference of neutralizing antibody titer level after dogs immuned rabies vaccine and determin whether the dogs need to be immuned again.
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Anticorpos Antivirais/sangue , Vacina Antirrábica/imunologia , Raiva/veterinária , Fitas Reagentes , Animais , Anticorpos Neutralizantes/análise , Anticorpos Neutralizantes/sangue , Anticorpos Antivirais/análise , Cromatografia de Afinidade/métodos , Cães , Coloide de Ouro , Raiva/prevenção & controle , Vírus da Raiva/imunologia , Sensibilidade e Especificidade , VacinaçãoRESUMO
An immunochromatographic test strip (ICTS) for detecting antibodies to rabies virus was developed, using colloidal gold particles labeled with rabies virus glycoprotein as the tracer. The assay was evaluated using sera from dogs immunized with various commercial rabies vaccines, or from dogs in the clinics and sera from dogs immunized with vaccines against pathogens other than rabies virus, and negative sera from a wide variety of animal sources, including dogs, mice, and cats which had never been vaccinated. The ICTS was found to be highly specific for antibodies against rabies virus, with a detection limit of 0.5IU/ml as measured by the fluorescent antibody virus neutralization (FAVN) test. Compared with the FAVN test, the specificity and sensitivity of ICTS were 98.2% and 90.4%, respectively. There was an excellent agreement between results obtained by the ICTS and FAVN tests (kappa=0.888). Strips stored at 4°C in a plastic bag with a desiccant retained their specificity and sensitivity for at least 15 months, and strips stored at ambient temperature remained stable for 12 months. The immunochromatographic test strip may therefore be useful for clinical laboratories lacking specialized equipment and for diagnosis in the field for rapid detection of rabies virus-specific antibodies.
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Anticorpos Antivirais/sangue , Imunoensaio/métodos , Vírus da Raiva/imunologia , Raiva/veterinária , Animais , Western Blotting , Gatos , Cães , Eletroforese em Gel de Poliacrilamida , Imunofluorescência , Glicoproteínas/imunologia , Coloide de Ouro , Camundongos , Testes de Neutralização , Raiva/diagnóstico , Raiva/virologia , Vacina Antirrábica/imunologia , Fitas Reagentes , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Proteínas Virais/imunologiaRESUMO
Nelson Bay orthoreovirus (NBV) is a species in the genus Orthoreovirus, family Reoviridae, containing 4, possibly 5, members. Here, we report a putative sixth member, Xi River virus (XRV), isolated from fruit bats collected in a location near the Xi River, Guangdong Province, China. This virus showed the same electron microscopic morphology as NBV, fusogenic CPE, and a 10-segmented double-strand RNA genome, as well as high sequence identity to NBV members. It is the first bat reovirus isolated in China.
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Quirópteros/virologia , Orthoreovirus , Animais , Sequência de Bases , China , Efeito Citopatogênico Viral , Pulmão/virologia , Dados de Sequência Molecular , Orthoreovirus/classificação , Orthoreovirus/genética , Orthoreovirus/isolamento & purificação , Orthoreovirus/patogenicidade , Filogenia , RNA de Cadeia Dupla/genética , RNA Viral/genética , Análise de Sequência de DNARESUMO
The plaque-forming characteristics of Newcastle disease viruses of chickens and geese source were compared on various cells. The result showed that there were obvious differences of plaque formation between F48E9 and NA-1 on Vero cells, chicken embryo fibroblast cells (CEF) and goose embryo fibroblast cells (GEF). The plaque-forming ability of NA-1 was higher than F48E9 on GEF, but lower than F48E9 on CEF. On Vero cells, the plaque-forming ability of NA-1 was slightly stronger than F48E9. It demonstrated that the plaque-forming characteristics were consistent with host tropism of virus. The morphogenesis of F48E9 and NA-1 on Vero cells was observed with transmission electron microscope. There were different replication processes between F48E9 and NA-1 on cells at different stages. NA-1 had stronger adaptability to host than F48E9 according to budding processes and envelope integrity.
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Galinhas , Gansos , Interações Hospedeiro-Patógeno , Doença de Newcastle/virologia , Vírus da Doença de Newcastle/crescimento & desenvolvimento , Vírus da Doença de Newcastle/ultraestrutura , Doenças das Aves Domésticas/virologia , Animais , Embrião de Galinha , Chlorocebus aethiops , Vírus da Doença de Newcastle/isolamento & purificação , Vírus da Doença de Newcastle/fisiologia , Células Vero , Ensaio de Placa ViralRESUMO
A feline panleukopenia virus (FPV) mutant, monkey/BJ-22/2008/CHN, was isolated from intestinal contents of a diarrheic monkey in Beijing, China. The virus was identified by morphology and physicochemical characteristics, and specific fragments were obtained by PCR using consensus primers of parvovirus and specific primers of FPV. Sequence of the full-length VP2 gene of the isolated FPV was determined and analyzed by comparison with reference FPV and canine parvovirus (CPV) isolates, showing high homology with FPV (98.75%) and CPV (98.15%). Phylogenetic analysis indicated that the isolated FPV formed a monophyletic branch in FPV cluster which differed from the other 11 FPV isolates from China and other countries. The isolated virus caused typical clinical symptoms of FPV in cats. This is the first report on isolation of FPV from a monkey.
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Diarreia/veterinária , Surtos de Doenças/veterinária , Vírus da Panleucopenia Felina/genética , Doenças dos Macacos/virologia , Infecções por Parvoviridae/veterinária , Animais , Gatos , China/epidemiologia , Diarreia/virologia , Panleucopenia Felina/virologia , Vírus da Panleucopenia Felina/isolamento & purificação , Feminino , Masculino , Doenças dos Macacos/epidemiologia , Doenças dos Macacos/patologia , Infecções por Parvoviridae/virologia , FilogeniaRESUMO
Six recombinant proteins GST-A beta 28/A beta 35/A beta 42 and GST-I-A beta 28/A beta 35/A beta 42 [I was the abbreviation for an immunostimulatory sequence that consisted of pan HLA DR binding epitope (PADRE) and Tetanus toxin epitope (TT)] were used as antigens after expressed and purified to immunize mice. The strongest antibody response against A beta 42 (titer 1:3200) was achieved by GST-I-A beta 28 or GST-A beta 42 immunization. However, IgG1 and IgG2b were the predominant serum antibody isotype responses by GST-I-A beta 28 immunization, whereas did IgG2a by GST-A beta 42 immunization. Thus, it indicated that GST-I-A beta 28 immunization in a mouse mainly evoked a stronger Th-2-type response; whereas, GST-A beta 42 immunization mainly elicited a Th-1-type response. Moreover, GST-I-A beta 28-induced serum antibodies had higher specificity to A beta 42 monomers and oligomers than to protofibrils and mature fibrils and exhibited the highest efficacy to block A beta 42 aggregation or fibrillogenesis and to disassemble A beta 42 aggregates in vitro. GST-I-A beta 28-induced serum antibodies also showed the most protective and restorative effects on target cells in vitro by inhibiting or neutralizing A beta 42-induced cytotoxicity. All of the above results indicated that A beta 28 could be speculated to substitute for A beta 42 and would become a better antigenic peptide for Alzheimer's disease immunotherapy in the presence of additional Th-cell epitopes such as the immunostimulatory sequence (I) applied in this study.
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Peptídeos beta-Amiloides/antagonistas & inibidores , Peptídeos beta-Amiloides/imunologia , Fragmentos de Peptídeos/antagonistas & inibidores , Fragmentos de Peptídeos/imunologia , Proteínas Recombinantes de Fusão/farmacologia , Doença de Alzheimer/tratamento farmacológico , Doença de Alzheimer/imunologia , Animais , Anticorpos/sangue , Epitopos/imunologia , Imunização/métodos , Imunoglobulina G/sangue , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Fragmentos de Peptídeos/farmacologia , Fragmentos de Peptídeos/uso terapêutico , Placa Amiloide/efeitos dos fármacos , Placa Amiloide/imunologia , Placa Amiloide/metabolismo , Proteínas Recombinantes de Fusão/imunologia , Proteínas Recombinantes de Fusão/uso terapêutico , Células Th1/efeitos dos fármacos , Células Th1/imunologia , Células Th2/efeitos dos fármacos , Células Th2/imunologiaRESUMO
Two giant pandas (Ailuropoda melanoleuca) died of unknown causes in a Chinese zoo. The clinical disease profile suggested that the pandas may have suffered a viral infection. Therefore, a series of detection including virus isolation, electron microscopy, cytobiological assay, serum neutralization and RT-PCR were used to identify the virus. It was determined that the isolated virus was a canine coronavirus (CCV), on the basis of coronavirus, neutralization by canine anti-CCV serum, and 84.3% to 100% amino acid sequence similarity with CCV. The results suggest that the affected pandas had been infected with CCV.
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Doenças dos Animais/virologia , Animais de Zoológico/virologia , Infecções por Coronaviridae/veterinária , Coronavirus Canino/isolamento & purificação , Ursidae/virologia , Sequência de Aminoácidos , Animais , Infecções por Coronaviridae/virologia , Coronavirus Canino/genética , Evolução Fatal , Feminino , Masculino , Dados de Sequência Molecular , Alinhamento de Sequência , Homologia de Sequência de Aminoácidos , Proteínas Virais/químicaRESUMO
Giardia canis virus (GCV) is a double-stranded RNA (dsRNA) virus of the family Totiviridae. In this study, the full length cDNA of the G. canis virus was constructed in pPoly2/sfinot vector and RNA was transcribed in vitro. Virus-free G. canis trophozoites were transfected with in vitro transcribed GCV RNA by electroporation. Transfected trophozoites were cultured for 12, 24, 36, 48, 60, or 72h post transfection for analysis. The ultrastructures of the transfected trophozoites were determined by transmission electron microscopy. The viral particles were detectable sporadically in the cytoplasm as early as 24h post transfection, but became evident and wide-spread 36h post transfection. The number of viral particles increased dramatically from 48 to 60h. Viral particles were released into the culture medium starting at about 60h and detectable in nuclei 72h post transfection. Severe vacuolization was seen in transfected G. canis trophozoites as early as 36h post transfection and persisted throughout the course of this study. The results of the present study indicate that in vitro transcribed GCV transcripts were capable of infecting Giardia trophozoites, apparently replicated and packaged into mature infectious viral particles which were released from the host.