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1.
mSphere ; 6(6): e0082021, 2021 12 22.
Artigo em Inglês | MEDLINE | ID: mdl-34935443

RESUMO

The upper respiratory tract is the primary site of infection by porcine hemagglutinating encephalomyelitis virus (PHEV). In this study, primary porcine respiratory epithelial cells (PRECs) were cultured in an air-liquid interface (ALI) to differentiate into a pseudostratified columnar epithelium, proliferative basal cells, M cells, ciliated cells, and mucus-secreting goblet cells. ALI-PRECs recreates a cell culture environment morphologically and functionally more representative of the epithelial lining of the swine trachea than traditional culture systems. PHEV replicated actively in this environment, inducing cytopathic changes and progressive disruption of the mucociliary apparatus. The innate immunity against PHEV was comparatively evaluated in ALI-PREC cultures and tracheal tissue sections derived from the same cesarean-derived, colostrum-deprived (CDCD) neonatal donor pigs. Increased expression levels of TLR3 and/or TLR7, RIG1, and MyD88 genes were detected in response to infection, resulting in the transcriptional upregulation of IFN-λ1 in both ALI-PREC cultures and tracheal epithelia. IFN-λ1 triggered the upregulation of the transcription factor STAT1, which in turn induced the expression of the antiviral IFN-stimulated genes OAS1 and Mx1. No significant modulation of the major proinflammatory cytokines interleukin-1ß (IL-1ß), IL-6, and tumor necrosis factor alpha (TNF-α) was detected in response to PHEV infection. However, a significant upregulation of different chemokines was observed in ALI-PREC cultures (CCL2, CCL5, CXCL8, and CXCL10) and tracheal epithelium (CXCL8 and CXCL10). This study shed light on the molecular mechanisms driving the innate immune response to PHEV at the airway epithelium, underscoring the important role of respiratory epithelial cells in the maintenance of respiratory homeostasis and on the initiation, resolution, and outcome of the infectious process. IMPORTANCE The neurotropic betacoronavirus porcine hemagglutinating encephalomyelitis virus (PHEV) primarily infects and replicates in the swine upper respiratory tract, causing vomiting and wasting disease and/or encephalomyelitis in suckling pigs. This study investigated the modulation of key early innate immune genes at the respiratory epithelia in vivo, on tracheal tissue sections from experimentally infected pigs, and in vitro, on air-liquid interface porcine respiratory cell cultures. The results from the study underscore the important role of respiratory epithelial cells in maintaining respiratory homeostasis and on the initiation, resolution, and outcome of the PHEV infectious process.


Assuntos
Betacoronavirus 1/fisiologia , Interferons/genética , Interleucina-8/imunologia , Receptores de Reconhecimento de Padrão/genética , Mucosa Respiratória/imunologia , Mucosa Respiratória/virologia , Replicação Viral , Animais , Animais Recém-Nascidos , Betacoronavirus 1/imunologia , Infecções por Coronavirus/imunologia , Infecções por Coronavirus/virologia , Imunidade Inata/genética , Imunidade Inata/imunologia , Interferons/imunologia , Interleucina-8/genética , Mucosa Respiratória/patologia , Suínos , Regulação para Cima , Replicação Viral/imunologia
2.
J Virol ; 95(19): e0085121, 2021 09 09.
Artigo em Inglês | MEDLINE | ID: mdl-34287052

RESUMO

Uncoordinated 51-like kinase 1 (ULK1) is a well-characterized initiator of canonical autophagy under basal or pathological conditions. Porcine hemagglutinating encephalomyelitis virus (PHEV), a neurotropic betacoronavirus (ß-CoV), impairs ULK1 kinase but hijacks autophagy to facilitate viral proliferation. However, the machinery of PHEV-induced autophagy initiation upon ULK1 kinase deficiency remains unclear. Here, the time course of PHEV infection showed a significant accumulation of autophagosomes (APs) in nerve cells in vivo and in vitro. Utilizing ULK1-knockout neuroblastoma cells, we have identified that ULK1 is not essential for productive AP formation induced by PHEV. In vitro phosphorylation studies discovered that mTORC1-regulated ULK1 activation stalls during PHEV infection, whereas AP biogenesis was controlled by AMPK-driven BECN1 phosphorylation. A lack of BECN1 is sufficient to block LC3 lipidation and disrupt recruitment of the LC3-ATG14 complex. Moreover, BECN1 acts as a bona fide substrate for ULK1-independent neural autophagy, and ectopic expression of BECN1 somewhat enhances PHEV replication. These findings highlight a novel machinery of noncanonical autophagy independent of ULK1 that bypasses the conserved initiation circuit of AMPK-mTORC1-ULK1, providing new insights into the interplay between neurotropic ß-CoV and the host. IMPORTANCE The ongoing coronavirus disease 2019 (COVID-19) pandemic alongside the outbreaks of severe acute respiratory syndrome (SARS) and Middle East respiratory syndrome (MERS) pose Betacoronavirus (ß-CoV) as a global public health challenge. Coronaviruses subvert, hijack, or utilize autophagy to promote proliferation, and thus, exploring the cross talk between ß-CoV and autophagy is of great significance in confronting future ß-CoV outbreaks. Porcine hemagglutinating encephalomyelitis virus (PHEV) is a highly neurotropic ß-CoV that invades the central nervous system (CNS) in pigs, but understanding of the pathogenesis for PHEV-induced neurological dysfunction is yet limited. Here, we discovered a novel regulatory principle of neural autophagy initiation during PHEV infection, where productive autophagosome (AP) biogenesis bypasses the multifaceted regulation of ULK1 kinase. The PHEV-triggered noncanonical autophagy underscores the complex interactions of virus and host and will help in the development of therapeutic strategies targeting noncanonical autophagy to treat ß-CoV disease.


Assuntos
Proteína Homóloga à Proteína-1 Relacionada à Autofagia/genética , Proteína Homóloga à Proteína-1 Relacionada à Autofagia/metabolismo , Autofagia/fisiologia , Betacoronavirus 1/metabolismo , Animais , Autofagossomos/metabolismo , Proteína Beclina-1/metabolismo , COVID-19 , Linhagem Celular , Técnicas de Inativação de Genes , Peptídeos e Proteínas de Sinalização Intracelular/metabolismo , Masculino , Alvo Mecanístico do Complexo 1 de Rapamicina/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Neurônios/metabolismo , Fosforilação , SARS-CoV-2
3.
Comp Immunol Microbiol Infect Dis ; 77: 101668, 2021 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-34004508

RESUMO

In 2020, an outbreak of equine coronavirus (ECoV) infection occurred among 41 horses at a riding stable in Tokyo, Japan. This stable had 16 Thoroughbreds and 25 horses of other breeds, including Andalusians, ponies and miniature horses. Fifteen horses (37 %) showed mild clinical signs such as fever, lethargy, anorexia and diarrhoea, and they recovered within 3 days of onset. A virus neutralization test showed that all 41 horses were infected with ECoV, signifying that 26 horses (63 %) were subclinical. The results suggest that subclinical horses played an important role as spreaders. A genome sequence analysis revealed that the lengths from genes p4.7 to p12.7 or NS2 in ECoV differed from those of ECoVs detected previously, suggesting that this outbreak was caused by a virus different from those that caused previous outbreaks among draughthorses in Japan. Among 30 horses that tested positive by real-time RT-PCR, ECoV shedding periods of non-Thoroughbreds were significantly longer than those of Thoroughbreds. The difference in shedding periods may indicate that some breeds excrete ECoV longer than other breeds and can contribute to the spread of ECoV.


Assuntos
Betacoronavirus 1 , Infecções por Coronavirus , Doenças dos Cavalos , Animais , Infecções por Coronavirus/epidemiologia , Infecções por Coronavirus/veterinária , Surtos de Doenças/veterinária , Doenças dos Cavalos/epidemiologia , Cavalos , Japão/epidemiologia , Tóquio
4.
J Virol ; 95(12)2021 05 24.
Artigo em Inglês | MEDLINE | ID: mdl-33762411

RESUMO

Porcine hemagglutinating encephalomyelitis virus (PHEV) is a betacoronavirus that causes vomiting and wasting disease and/or encephalomyelitis in suckling pigs. This study characterized PHEV infection, pathogenesis, and immune response in cesarean-derived, colostrum-deprived (CDCD) neonatal pigs. Infected animals developed mild respiratory, enteric, and neurological clinical signs between 2 to 13 days postoronasal inoculation (dpi). PHEV did not produce viremia, but virus shedding was detected in nasal secretions (1 to 10 dpi) and feces (2 to 7 dpi) by reverse transcriptase quantitative PCR (RT-qPCR). Viral RNA was detected in all tissues except liver, but the detection rate and RT-qPCR threshold cycle (CT ) values decreased over time. The highest concentration of virus was detected in inoculated piglets necropsied at 5 dpi in turbinate and trachea, followed by tonsils, lungs, tracheobronchial lymph nodes, and stomach. The most representative microscopic lesions were gastritis lymphoplasmacytic, moderate, multifocal, with perivasculitis, and neuritis with ganglia degeneration. A moderate inflammatory response, characterized by increased levels of interferon alpha (IFN-α) in plasma (5 dpi) and infiltration of T lymphocytes and macrophages were also observed. Increased plasma levels of interleukin-8 (IL-8) were detected at 10 and 15 dpi, coinciding with the progressive resolution of the infection. Moreover, a robust antibody response was detected by 10 dpi. An ex vivo air-liquid CDCD-derived porcine respiratory cells culture (ALI-PRECs) system showed virus replication in ALI-PRECs and cytopathic changes and disruption of ciliated columnar epithelia, thereby confirming the tracheal epithelia as a primary site of infection for PHEV.IMPORTANCE Among the ∼46 virus species in the family Coronaviridae, many of which are important pathogens of humans and 6 of which are commonly found in pigs, porcine hemagglutinating encephalomyelitis remains one of the least researched. The present study provided a comprehensive characterization of the PHEV infection process and immune responses using CDCD neonatal pigs. Moreover, we used an ex vivo ALI-PRECs system resembling the epithelial lining of the tracheobronchial region of the porcine respiratory tract to demonstrate that the upper respiratory tract is a primary site of PHEV infection. This study provides a platform for further multidisciplinary studies of coronavirus infections.


Assuntos
Betacoronavirus 1/imunologia , Infecções por Coronavirus/imunologia , Interferon-alfa/imunologia , Interleucina-8/imunologia , Doenças dos Suínos/imunologia , Linfócitos T/imunologia , Animais , Linhagem Celular , Infecções por Coronavirus/patologia , Infecções por Coronavirus/veterinária , Especificidade de Órgãos/imunologia , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Suínos , Doenças dos Suínos/patologia , Linfócitos T/patologia , Linfócitos T/virologia
5.
Vet Microbiol ; 255: 109015, 2021 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-33640682

RESUMO

Porcine hemagglutinating encephalomyelitis virus (PHEV) displays neurotropism and induces atypical autophagy. However, the exact mechanisms mediating autophagy induced by PHEV remains uncharacterized. Transcription factor EB (TFEB) is a master transcriptional regulator playing a key role in autophagy and its activity is regulated by MTORC1 kinase on the surface of lysosomes. We first found that PHEV infection decreases TFEB expression, while it activates TFEB by inhibiting MTORC1 activation, indicating that TFEB plays a complex role in the process of PHEV-induced autophagy through opposite regulation of its expression and activity. Furthermore, this study preliminarily demonstrated that PHEV replication is dependent on TFEB expression.


Assuntos
Autofagia/fisiologia , Fatores de Transcrição de Zíper de Leucina e Hélice-Alça-Hélix Básicos/metabolismo , Betacoronavirus 1 , Regulação da Expressão Gênica , Glicoproteínas de Membrana Associadas ao Lisossomo/metabolismo , Replicação Viral/fisiologia , Transporte Ativo do Núcleo Celular , Animais , Fatores de Transcrição de Zíper de Leucina e Hélice-Alça-Hélix Básicos/genética , Linhagem Celular , Glicoproteínas de Membrana Associadas ao Lisossomo/genética , Alvo Mecanístico do Complexo 1 de Rapamicina/genética , Alvo Mecanístico do Complexo 1 de Rapamicina/metabolismo , Camundongos , Suínos
6.
Vet Microbiol ; 253: 108958, 2021 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-33387911

RESUMO

Porcine hemagglutinating encephalomyelitis virus (PHEV) is the cause of acute outbreaks of vomiting and wasting disease and/or encephalomyelitis in neonatal pigs, with naïve herds particularly vulnerable to clinical episodes. PHEV infections in older pigs are generally considered to be subclinical, but are poorly characterized in the refereed literature. In this study, twelve 7-week-old pigs were oronasally inoculated with 0.5 mL (1:128 HA titer) PHEV (Mengeling strain) and then followed through 42 days post inoculation (dpi). Fecal and oral fluid specimens were collected daily to evaluate viral shedding. Serum samples were tested for viremia, isotype-specific antibody responses, cytokine, and chemokine responses. Peripheral blood mononuclear cells were isolated to evaluate phenotype changes in immune cell subpopulations. No clinical signs were observed in PHEV inoculated pigs, but virus was detected in oral fluid (1-28 dpi) and feces (1-10 dpi). No viremia was detected, but a significant IFN-α response was observed in serum at 3 dpi, followed by the detection of IgM (dpi 7), and IgA/IgG (dpi 10). Flow cytometry revealed a one-off increase in cytotoxic T cells at 21 dpi. This study demonstrated that exposure of grower pigs to PHEV results in subclinical infection characterized by active viral replication and shedding followed by an active humoral and cell-mediated immune response that attenuates the course of the infection and results in viral clearance.


Assuntos
Betacoronavirus 1/isolamento & purificação , Infecções por Coronavirus/veterinária , Doenças dos Suínos/virologia , Animais , Anticorpos Antivirais/sangue , Infecções por Coronavirus/imunologia , Infecções por Coronavirus/virologia , Interferon-alfa/biossíntese , Interferon-alfa/sangue , Suínos , Doenças dos Suínos/sangue , Doenças dos Suínos/imunologia , Viremia
7.
J Virol Methods ; 289: 114016, 2021 03.
Artigo em Inglês | MEDLINE | ID: mdl-33290788

RESUMO

Porcine hemagglutinating encephalomyelitis virus (PHEV) is a member of the genus Betacoronavirus and is the etiologic agent of encephalomyelitis or vomiting and wasting disease in neonatal pigs. Although there are only a few epidemiological studies that document the seroprevalence of PHEV infection, there are reports of sporadic outbreaks, including recent documentation of an influenza-like respiratory disease associated with PHEV in the United States. To address this issue, we have developed a new indirect enzyme linked immunosorbent assay (ELISA) for use in sero-epidemiological research of PHEV infection. One hundred and fifty porcine serum samples that were determined as antibody-positive or antibody-negative in virus neutralization (VN) tests were used in conjunction with PHEV-specific antigen extracted from virus-infected FS-L3 cells using RBS buffer containing 0.2 % NP-40 to develop this assay. The ELISA showed a high sensitivity (95.35 %) and specificity (96.88 %) by receiver operating characteristic (ROC) analysis, with an area under the curve (AUC) of 0.996 attesting to its accuracy. Our results revealed a strong correlation between the results of the indirect ELISA and VN test (R = 0.850, P < 0.05), with near-perfect agreement (kappa value = 0.932). These results indicate that this new indirect ELISA might be useful for diagnosis and sero-epidemiological tracking of PHEV infection.


Assuntos
Anticorpos Antivirais/sangue , Betacoronavirus 1/imunologia , Infecções por Coronavirus/veterinária , Ensaio de Imunoadsorção Enzimática/métodos , Doenças dos Suínos/diagnóstico , Animais , Linhagem Celular , Infecções por Coronavirus/diagnóstico , Sensibilidade e Especificidade , Estudos Soroepidemiológicos , Suínos , Doenças dos Suínos/virologia
8.
Vet Microbiol ; 252: 108918, 2021 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-33191000

RESUMO

Porcine haemagglutinating encephalomyelitis virus (PHEV) is a member of coronavirus that causes acute infectious disease and high mortality in piglets. The transcription factor IRF3 is a central regulator of type I interferon (IFN) innate immune signalling. Here, we report that PHEV infection of RAW264.7 cells results in strong suppression of IFN-ß production in the early stage. A comparative analysis of the upstream effector of IFN-ß transcription demonstrated that deactivation of IRF3, but not p65 or ATF-2 proteins, is uniquely attributed to failure of early IFN-ß induction. Moreover, the RIG-I/MDA5/MAVS/TBK1-dependent protective response that regulates the IRF3 pathway is not disrupted by PHEV and works well underlying the deactivated IRF3-mediated IFN-ß inhibition. After challenge with poly(I:C), a synthetic analogue of dsRNA used to stimulate IFN-ß secretion in the TLR-controlled pathway, we show that PHEV and poly(I:C) regulate IFN-ß-induction via two different pathways. Collectively, our findings reveal that deactivation of IRF3 is a specific mechanism that contributes to termination of type I IFN signalling during early infection with PHEV independent of the conserved RIG-I/MAVS/MDA5/TBK1-mediated innate immune response.


Assuntos
Betacoronavirus 1/imunologia , Infecções por Coronavirus/veterinária , Fator Regulador 3 de Interferon/genética , Interferon beta/imunologia , Animais , Betacoronavirus 1/genética , Infecções por Coronavirus/imunologia , Infecções por Coronavirus/virologia , Imunidade Inata , Fator Regulador 3 de Interferon/imunologia , Camundongos , Poli I-C/farmacologia , Células RAW 264.7 , Transdução de Sinais/imunologia
9.
Mol Neurobiol ; 57(12): 5299-5306, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-32876841

RESUMO

Lysosomes are involved in pathogenesis of a variety of neurodegenerative diseases and play a large role in neurodegenerative disorders caused by virus infection. However, whether virus-infected cells or animals can be used as experimental models of neurodegeneration in humans based on virus-related lysosomal dysfunction remain unclear. Porcine hemagglutinating encephalomyelitis virus displays neurotropism in mice, and neural cells are its targets for viral progression. PHEV infection was confirmed to be a risk factor for neurodegenerative diseases in the present. The findings demonstrated for the first time that PHEV infection can lead to lysosome disorders and showed that the specific mechanism of lysosome dysfunction is related to PGRN expression deficiency and indicated similar pathogenesis compared with human neurodegenerative diseases upon PHEV infection. Trehalose can also increase progranulin expression and rescue abnormalities in lysosomal structure in PHEV-infected cells. In conclusion, these results suggest that PHEV probably serve as a disease model for studying the pathogenic mechanisms and prevention of other degenerative diseases.


Assuntos
Betacoronavirus 1/fisiologia , Lisossomos/patologia , Doenças Neurodegenerativas/patologia , Doenças Neurodegenerativas/virologia , Animais , Linhagem Celular Tumoral , Infecções por Coronavirus/patologia , Infecções por Coronavirus/virologia , Proteínas de Ligação a DNA/metabolismo , Modelos Animais de Doenças , Lisossomos/efeitos dos fármacos , Masculino , Camundongos Endogâmicos BALB C , Progranulinas/metabolismo , Suínos , Trealose/farmacologia
11.
mSphere ; 5(3)2020 05 06.
Artigo em Inglês | MEDLINE | ID: mdl-32376700

RESUMO

Members of family Coronaviridae cause a variety of diseases in birds and mammals. Porcine hemagglutinating encephalomyelitis virus (PHEV), a lesser-researched coronavirus, can infect naive pigs of any age, but clinical disease is observed in pigs ≤4 weeks of age. No commercial PHEV vaccines are available, and neonatal protection from PHEV-associated disease is presumably dependent on lactogenic immunity. Although subclinical PHEV infections are thought to be common, PHEV ecology in commercial swine herds is unknown. To begin to address this gap in knowledge, a serum IgG antibody enzyme-linked immunosorbent assay (ELISA) based on the S1 protein was developed and evaluated on known-status samples and then used to estimate PHEV seroprevalence in U.S. sow herds. Assessment of the diagnostic performance of the PHEV S1 ELISA using serum samples (n = 924) collected from 7-week-old pigs (n = 84; 12 pigs per group) inoculated with PHEV, porcine epidemic diarrhea virus, transmissible gastroenteritis virus, porcine respiratory coronavirus, or porcine deltacoronavirus showed that a sample-to-positive cutoff value of ≥0.6 was both sensitive and specific, i.e., all PHEV-inoculated pigs were seropositive from days postinoculation 10 to 42, and no cross-reactivity was observed in samples from other groups. The PHEV S1 ELISA was then used to estimate PHEV seroprevalence in U.S. sow herds (19 states) using 2,756 serum samples from breeding females (>28 weeks old) on commercial farms (n = 104) with no history of PHEV-associated disease. The overall seroprevalence was 53.35% (confidence interval [CI], ±1.86%) and herd seroprevalence was 96.15% (CI, ±3.70%).IMPORTANCE There is a paucity of information concerning the ecology of porcine hemagglutinating encephalomyelitis virus (PHEV) in commercial swine herds. This study provided evidence that PHEV infection is endemic and highly prevalent in U.S. swine herds. These results raised questions for future studies regarding the impact of endemic PHEV on swine health and the mechanisms by which this virus circulates in endemically infected populations. Regardless, the availability of the validated PHEV S1 enzyme-linked immunosorbent assay (ELISA) provides the means for swine producers to detect and monitor PHEV infections, confirm prior exposure to the virus, and to evaluate the immune status of breeding herds.


Assuntos
Anticorpos Antivirais/sangue , Betacoronavirus 1/isolamento & purificação , Infecções por Coronavirus/epidemiologia , Infecções por Coronavirus/veterinária , Doenças dos Suínos/epidemiologia , Animais , Anticorpos Antivirais/imunologia , Betacoronavirus 1/imunologia , Infecções por Coronavirus/diagnóstico , Reações Cruzadas/imunologia , Ensaio de Imunoadsorção Enzimática/métodos , Imunoglobulina G/sangue , Imunoglobulina G/imunologia , Coronavirus Respiratório Porcino/imunologia , Vírus da Diarreia Epidêmica Suína/imunologia , Estudos Soroepidemiológicos , Suínos , Doenças dos Suínos/diagnóstico , Vírus da Gastroenterite Transmissível/imunologia , Estados Unidos/epidemiologia
12.
J Equine Vet Sci ; 87: 102906, 2020 04.
Artigo em Inglês | MEDLINE | ID: mdl-32172908

RESUMO

Equine coronavirus (ECoV) is a known cause of fever, anorexia, and lethargy in adult horses. Although there are multiple reports of ECoV outbreaks, less is known about the clinical presentation of individual horses during a nonoutbreak situation. The purpose of this study was to describe the clinical presentation of horses diagnosed with ECoV infection that were not associated with an outbreak. Medical records of all horses admitted to Washington State University, Veterinary Teaching Hospital, during an 8-year period were reviewed (2010-2018). The five horses included in this study were older than 1 year of age, were diagnosed with colitis, tested positive for ECoV using real-time polymerase chain reaction, and were negative to other enteric pathogens. Interestingly, 4 of 5 horses had moderate to severe diarrhea, 3 had abnormal large colon ultrasonography, 2 had transient ventricular tachycardia and 2 had clinicopathologic evidence of liver dysfunction. ECoV should be included as a differential diagnosis for individual horses presenting with anorexia, fever, lethargy, and colitis. Early identification of ECoV cases is key to implement appropriate biosecurity measures to prevent the potential spread of this disease.


Assuntos
Betacoronavirus 1 , Colite/veterinária , Doenças dos Cavalos , Animais , Cavalos , Estudos Retrospectivos , Washington
13.
Arch Virol ; 165(2): 345-354, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31834525

RESUMO

Porcine hemagglutinating encephalomyelitis virus (PHEV) is a typical neurotropic coronavirus that mainly invades the central nervous system (CNS) in piglets and causes vomiting and wasting disease. Emerging evidence suggests that PHEV alters microRNA (miRNA) expression profiles, and miRNA has also been postulated to be involved in its pathogenesis, but the mechanisms underlying this process have not been fully explored. In this study, we found that PHEV infection upregulates miR-142a-3p RNA expression in N2a cells and in the CNS of mice. Downregulation of miR-142a-3p by an miRNA inhibitor led to a significant repression of viral proliferation, implying that it acts as a positive regulator of PHEV proliferation. Using a dual-luciferase reporter assay, miR-142a-3p was found to bind directly bound to the 3' untranslated region (3'UTR) of Rab3a mRNA and downregulate its expression. Knockdown of Rab3a expression by transfection with an miR-142a-3p mimic or Rab3a siRNA significantly increased PHEV replication in N2a cells. Conversely, the use of an miR-142a-3p inhibitor or overexpression of Rab3a resulted in a marked restriction of viral production at both the mRNA and protein level. Our data demonstrate that miR-142a-3p promotes PHEV proliferation by directly targeting Rab3a mRNA, and this provides new insights into the mechanisms of PHEV-related pathogenesis and virus-host interactions.


Assuntos
Betacoronavirus 1/genética , Proliferação de Células/genética , Infecções por Coronavirus/genética , MicroRNAs/genética , Suínos/virologia , Proteína rab3A de Ligação ao GTP/genética , Regiões 3' não Traduzidas/genética , Animais , Linhagem Celular , Linhagem Celular Tumoral , Infecções por Coronavirus/veterinária , Infecções por Coronavirus/virologia , Regulação para Baixo/genética , Células HEK293 , Humanos , Camundongos , RNA Mensageiro/genética , RNA Interferente Pequeno/genética , Regulação para Cima/genética
14.
Ciênc. Saúde Colet ; 25(9): 3567-3571, Mar. 2020.
Artigo em Português | LILACS, Coleciona SUS, Sec. Est. Saúde SP | ID: biblio-1133151

RESUMO

Resumo O Ministério da Saúde declarou em 03 de fevereiro de 2020 estado de emergência em saúde pública de importância nacional em decorrência da pandemia pelo novo coronavírus SARS-CoV-2. Com isso, o IBGE adiou a realização do Censo Demográfico de 2020 e passou a formular uma PNAD COVID-19. O inquérito contou com uma amostra total de 349 mil pessoas em cerca de 200 mil domicílios. Do total da população-residente brasileira, o IBGE estimou em maio/2020 que 24,0 milhões (11,4%) tiveram pelo menos um dos sintomas de síndrome gripal (SG). Desse contingente, 20,2 milhões (84,3% do total dos sintomáticos) não procuraram unidade de saúde. As inovações trazidas para a vigilância em saúde e o pioneirismo do IBGE demonstram ser possível, em um país continental e que vem experimentando diversas epidemias locais em momentos diferentes em seu território, que outros países também desenvolvam inquéritos domiciliares semelhantes, com coleta de dados semanal (referida às semanas epidemiológicas) por telefone de forma inovadora e tempestiva. A PNAD COVID-19 trouxe ainda uma nova tecnologia para o Instituto, resgatando o papel de avaliador externo do Sistema Único de Saúde (SUS).


Abstract On February 3, 2020, the Brazilian Ministry of Health declared a state of emergency in public health of national relevance due to the pandemic caused by the new coronavirus SARS-CoV-2. As a result, IBGE postponed the 2020 Demographic Census and started to formulate a COVID-19 PNAD. The survey included a total sample of 349 thousand people in about 200 thousand households. Of the total Brazilian resident population, the IBGE estimated in May/2020 that 24.0 million (11.4%) had at least one of the flu-like syndrome symptoms. Of this contingent, 20.2 million (84.3% of all symptomatic patients) did not seek health care. The innovations brought to health surveillance and the IBGE's pioneering spirit show that it is possible, in a continental country that has been experiencing several local epidemics at different times in its territory, that other countries also develop similar household surveys, with weekly data collection (referred to epidemiological weeks) by telephone in an innovative and timely manner. The COVID-19 PNAD also brought new technology to the Institute, reviving its role as an external evaluator of the Unified Health System (SUS).


Assuntos
Humanos , Pneumonia Viral/epidemiologia , Saúde Pública , Inquéritos e Questionários , Infecções por Coronavirus/epidemiologia , Vigilância em Saúde Pública/métodos , Pneumonia Viral/terapia , Pneumonia Viral/virologia , Brasil , Aceitação pelo Paciente de Cuidados de Saúde/estatística & dados numéricos , Infecções por Coronavirus , Infecções por Coronavirus/terapia , Infecções por Coronavirus/virologia , Tecnologia Biomédica , Atenção à Saúde/organização & administração , Pandemias , Betacoronavirus , Betacoronavirus 1/isolamento & purificação
17.
Artigo em Espanhol | LILACS | ID: biblio-1100478

RESUMO

Lay bioethics is an applied ethic that allows us to face the problems generated by the tension between science and the human being.The current pandemic caused by an unknown infectious agent has highlighted the need for the application of this applied ethic, not located on the laurels of the ethereal or purely intellectual, but rather based in day-to-day reality. Decision-making from all areas of health care requires this fundamental frame of reference; the application of correct precepts in epidemiological, sanitary decisions and political measures cannot and should not be done without this frame of reference; or the result will be catastrophic.


La Bioética laica es en principio una ética aplicada que permite afrontar los problemas generados por la tensión existente entre la ciencia y el ser humano. La actual pandemia causada por un agente infeccioso no conocido ha puesto de relieve la necesidad de la aplicación de esta ética aplicada, no ubicada en los laureles de lo etéreo o puramente intelectual sino más bien asentada en la realidad del día a día. La toma de decisiones desde todos los ámbitos del quehacer sanitario requiere este marco referencial fundamental; aplicación de correctos preceptos en las decisiones epidemiológicas, sanitarias y medidas políticas no puede ni debe hacerse sin este marco referencial; otrora el resultado será catastrófico.(AU)


Assuntos
Humanos , Saúde Global/estatística & dados numéricos , Temas Bioéticos/normas , Betacoronavirus 1/imunologia , Bioética , Regulamento Sanitário Internacional/estatística & dados numéricos
18.
Viruses ; 11(12)2019 11 30.
Artigo em Inglês | MEDLINE | ID: mdl-31801275

RESUMO

Equine coronavirus (ECoV) is considered to be involved in enteric diseases in foals. Recently, several outbreaks of ECoV infection have also been reported in adult horses from the USA, France and Japan. Epidemiological studies of ECoV infection are still limited, and the seroprevalence of ECoV infection in Europe is unknown. In this study, an indirect enzyme-linked immunosorbent assay (ELISA) method utilizing ECoV spike S1 protein was developed in two formats, and further validated by analyzing 27 paired serum samples (acute and convalescent sera) from horses involved in an ECoV outbreak and 1084 sera of horses with unknown ECoV exposure. Both formats showed high diagnostic accuracy compared to virus neutralization (VN) assay. Receiver-operating characteristic (ROC) analyses were performed to determine the best cut-off values for both ELISA formats, assuming a test specificity of 99%. Employing the developed ELISA method, we detected seroconversion in 70.4% of horses from an ECoV outbreak. Among the 1084 horse sera, seropositivity varied from 25.9% (young horses) to 82.8% (adult horses) in Dutch horse populations. Further, sera of Icelandic horses were included in this study and a significant number of sera (62%) were found to be positive. Overall, the results demonstrated that the ECoV S1-based ELISA has reliable diagnostic performance compared to the VN assay and is a useful assay to support seroconversion in horses involved with ECoV outbreaks and to estimate ECoV seroprevalence in populations of horses.


Assuntos
Anticorpos Antivirais/sangue , Betacoronavirus 1/isolamento & purificação , Infecções por Coronavirus/veterinária , Doenças dos Cavalos/diagnóstico , Testes Sorológicos/veterinária , Glicoproteína da Espícula de Coronavírus/imunologia , Animais , Anticorpos Neutralizantes/sangue , Betacoronavirus 1/imunologia , Infecções por Coronavirus/diagnóstico , Infecções por Coronavirus/epidemiologia , Ensaio de Imunoadsorção Enzimática/veterinária , Doenças dos Cavalos/epidemiologia , Cavalos , Islândia/epidemiologia , Países Baixos/epidemiologia , Curva ROC , Estudos Soroepidemiológicos , Testes Sorológicos/métodos
19.
Viruses ; 11(10)2019 10 14.
Artigo em Inglês | MEDLINE | ID: mdl-31615132

RESUMO

The objective of this study was to investigate the presence of equine coronavirus (ECoV) in clinical samples submitted to a diagnostic laboratory in Ireland. A total of 424 clinical samples were examined from equids with enteric disease in 24 Irish counties between 2011 and 2015. A real-time reverse transcription polymerase chain reaction was used to detect ECoV RNA. Nucleocapsid, spike and the region from the p4.7 to p12.7 genes of positive samples were sequenced, and sequence and phylogenetic analyses were conducted. Five samples (1.2%) collected in 2011 and 2013 tested positive for ECoV. Positive samples were collected from adult horses, Thoroughbred foals and a donkey foal. Sequence and/or phylogenetic analysis showed that nucleocapsid, spike and p12.7 genes were highly conserved and were closely related to ECoVs identified in other countries. In contrast, the region from p4.7 and the non-coding region following the p4.7 gene had deletions or insertions. The differences in the p4.7 region between the Irish ECoVs and other ECoVs indicated that the Irish viruses were distinguishable from those circulating in other countries. This is the first report of ECoV detected in both foals and adult horses in Ireland.


Assuntos
Betacoronavirus 1/genética , Infecções por Coronavirus/veterinária , Doenças dos Cavalos/epidemiologia , Filogenia , Animais , Betacoronavirus 1/classificação , Infecções por Coronavirus/epidemiologia , DNA Viral/genética , Fezes/virologia , Doenças dos Cavalos/diagnóstico , Doenças dos Cavalos/virologia , Cavalos , Irlanda/epidemiologia , Análise de Sequência de DNA , Deleção de Sequência
20.
Vet Res ; 50(1): 63, 2019 Sep 18.
Artigo em Inglês | MEDLINE | ID: mdl-31533860

RESUMO

Widespread geographic movement and extensive comingling of exhibition swine facilitates the spread and transmission of infectious pathogens. Nasal samples were collected from 2862 pigs at 102 exhibitions and tested for five pathogens. At least one pathogen was molecularly detected in pigs at 63 (61.8%) exhibitions. Influenza A virus was most prevalent and was detected in 498 (17.4%) samples. Influenza D virus was detected in two (0.07%) samples. More than one pathogen was detected in 165 (5.8%) samples. Influenza A virus remains a top threat to animal and human health, but other pathogens may be disseminated through the exhibition swine population.


Assuntos
Doenças Respiratórias/veterinária , Doenças dos Suínos/epidemiologia , Animais , Betacoronavirus 1/isolamento & purificação , Infecções por Coronavirus/epidemiologia , Infecções por Coronavirus/veterinária , Infecções por Coronavirus/virologia , Vírus da Influenza A/isolamento & purificação , Infecções por Orthomyxoviridae/epidemiologia , Infecções por Orthomyxoviridae/veterinária , Infecções por Orthomyxoviridae/virologia , Síndrome Respiratória e Reprodutiva Suína/epidemiologia , Síndrome Respiratória e Reprodutiva Suína/virologia , Vírus da Síndrome Respiratória e Reprodutiva Suína/isolamento & purificação , Prevalência , Doenças Respiratórias/epidemiologia , Doenças Respiratórias/virologia , Respirovirus/isolamento & purificação , Infecções por Respirovirus/epidemiologia , Infecções por Respirovirus/veterinária , Infecções por Respirovirus/virologia , Sus scrofa , Suínos , Doenças dos Suínos/virologia , Thogotovirus/isolamento & purificação , Estados Unidos/epidemiologia
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