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1.
Int J Mol Sci ; 24(8)2023 Apr 14.
Artículo en Inglés | MEDLINE | ID: mdl-37108459

RESUMEN

Acute respiratory distress syndrome is an acute respiratory failure caused by cytokine storms; highly pathogenic influenza A virus infection can induce cytokine storms. The innate immune response is vital in this cytokine storm, acting by activating the transcription factor NF-κB. Tissue injury releases a danger-associated molecular pattern that provides positive feedback for NF-κB activation. Exogenous mesenchymal stem cells can also modulate immune responses by producing potent immunosuppressive substances, such as prostaglandin E2. Prostaglandin E2 is a critical mediator that regulates various physiological and pathological processes through autocrine or paracrine mechanisms. Activation of prostaglandin E2 results in the accumulation of unphosphorylated ß-catenin in the cytoplasm, which subsequently reaches the nucleus to inhibit the transcription factor NF-κB. The inhibition of NF-κB by ß-catenin is a mechanism that reduces inflammation.


Asunto(s)
Gripe Humana , Síndrome de Dificultad Respiratoria , Humanos , FN-kappa B/metabolismo , Dinoprostona , beta Catenina/metabolismo , Síndrome de Liberación de Citoquinas , Vía de Señalización Wnt , Inmunidad Innata
2.
Hum Genomics ; 15(1): 29, 2021 05 17.
Artículo en Inglés | MEDLINE | ID: mdl-34001248

RESUMEN

BACKGROUND: Coronavirus disease 2019 (COVID-19) is a global health problem that causes millions of deaths worldwide. The clinical manifestation of COVID-19 widely varies from asymptomatic infection to severe pneumonia and systemic inflammatory disease. It is thought that host genetic variability may affect the host's response to the virus infection and thus cause severity of the disease. The SARS-CoV-2 virus requires interaction with its receptor complex in the host cells before infection. The transmembrane protease serine 2 (TMPRSS2) has been identified as one of the key molecules involved in SARS-CoV-2 virus receptor binding and cell invasion. Therefore, in this study, we investigated the correlation between a genetic variant within the human TMPRSS2 gene and COVID-19 severity and viral load. RESULTS: We genotyped 95 patients with COVID-19 hospitalised in Dr Soetomo General Hospital and Indrapura Field Hospital (Surabaya, Indonesia) for the TMPRSS2 p.Val160Met polymorphism. Polymorphism was detected using a TaqMan assay. We then analysed the association between the presence of the genetic variant and disease severity and viral load. We did not observe any correlation between the presence of TMPRSS2 genetic variant and the severity of the disease. However, we identified a significant association between the p.Val160Met polymorphism and the SARS-CoV-2 viral load, as estimated by the Ct value of the diagnostic nucleic acid amplification test. Furthermore, we observed a trend of association between the presence of the C allele and the mortality rate in patients with severe COVID-19. CONCLUSION: Our data indicate a possible association between TMPRSS2 p.Val160Met polymorphism and SARS-CoV-2 infectivity and the outcome of COVID-19.


Asunto(s)
COVID-19/genética , Predisposición Genética a la Enfermedad/genética , Polimorfismo de Nucleótido Simple , SARS-CoV-2/aislamiento & purificación , Serina Endopeptidasas/genética , Adulto , Alelos , COVID-19/diagnóstico , COVID-19/virología , Estudios Transversales , Femenino , Frecuencia de los Genes , Genotipo , Humanos , Indonesia , Masculino , Persona de Mediana Edad , SARS-CoV-2/fisiología , Carga Viral/genética
3.
BMC Infect Dis ; 20(1): 823, 2020 Nov 11.
Artículo en Inglés | MEDLINE | ID: mdl-33176722

RESUMEN

BACKGROUND: The highly pathogenic avian influenza A/H5N1 virus is one of the causative agents of acute lung injury (ALI) with high mortality rate. Studies on therapeutic administration of bone marrow-derived mesenchymal stem cells (MSCs) in ALI caused by the viral infection have been limited in number and have shown conflicting results. The aim of the present investigation is to evaluate the therapeutic potential of MSC administration in A/H5N1-caused ALI, using a mouse model. METHODS: MSCs were prepared from the bone marrow of 9 to 12 week-old BALB/c mice. An H5N1 virus of A/turkey/East Java/Av154/2013 was intranasally inoculated into BALB/c mice. On days 2, 4, and 6 after virus inoculation, MSCs were intravenously administered into the mice. To evaluate effects of the treatment, we examined for lung alveolar protein as an indicator for lung injury, PaO2/FiO2 ratio for lung functioning, and lung histopathology. Expressions of NF-κB, RAGE (transmembrane receptor for damage associated molecular patterns), TNFα, IL-1ß, Sftpc (alveolar cell type II marker), and Aqp5+ (alveolar cell type I marker) were examined by immunohistochemistry. In addition, body weight, virus growth in lung and brain, and duration of survival were measured. RESULTS: The administration of MSCs lowered the level of lung damage in the virus-infected mice, as shown by measuring lung alveolar protein, PaO2/FiO2 ratio, and histopathological score. In the MSC-treated group, the expressions of NF-κB, RAGE, TNFα, and IL-1ß were significantly suppressed in comparison with a mock-treated group, while those of Sftpc and Aqp5+ were enhanced. Body weight, virus growth, and survival period were not significantly different between the groups. CONCLUSION: The administration of MSCs prevented further lung injury and inflammation, and enhanced alveolar cell type II and I regeneration, while it did not significantly affect viral proliferation and mouse morbidity and mortality. The results suggested that MSC administration was a promissing strategy for treatment of acute lung injuries caused by the highly pathogenic avian influenza A/H5N1 virus, although further optimization and combination use of anti-viral drugs will be obviously required to achieve the goal of reducing mortality.


Asunto(s)
Lesión Pulmonar Aguda/etiología , Lesión Pulmonar Aguda/cirugía , Subtipo H5N1 del Virus de la Influenza A , Trasplante de Células Madre Mesenquimatosas , Infecciones por Orthomyxoviridae/complicaciones , Neumonía/etiología , Neumonía/cirugía , Lesión Pulmonar Aguda/prevención & control , Lesión Pulmonar Aguda/virología , Animales , Citocinas/metabolismo , Modelos Animales de Enfermedad , Pulmón/metabolismo , Pulmón/virología , Masculino , Ratones , Ratones Endogámicos BALB C , Infecciones por Orthomyxoviridae/virología , Neumonía/prevención & control , Neumonía/virología , Resultado del Tratamiento
4.
Arch Virol ; 163(8): 2099-2109, 2018 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-29633076

RESUMEN

We evaluated the anti-influenza-virus effects of Melia components and discuss the utility of these components. The effects of leaf components of Melia azedarach L. on viruses were examined, and plaque inhibition tests were performed. The in vivo efficacy of M. azedarach L. was tested in a mouse model. Leaf components of Melia azedarach L. markedly inhibited the growth of various influenza viruses. In an initial screening, multiplication and haemagglutination (HA) activities of H1N1, H3N2, H5, and B influenza viruses were inactivated by the liquid extract of leaves of M. azedarach L. (MLE). Furthermore, plaque inhibition titres of H1N1, H3N2, and B influenza viruses treated with MLE ranged from 103.7 to 104.2. MLE possessed high plaque-inhibitory activity against pandemic avian H5N1, H7N9, and H9N2 vaccine candidate strains, with a plaque inhibition titre of more than 104.2. Notably, the buoyant density decreased from 1.175 to 1.137 g/cm3, and spikeless particles appeared. We identified four anti-influenza virus substances: pheophorbide b, pheophorbide a, pyropheophorbide a, and pheophytin a. Photomorphogenesis inside the envelope may lead to removal of HA and neuraminidase spikes from viruses. Thus, MLE could efficiently remove floating influenza virus in the air space without toxicity. Consistent with this finding, intranasal administration of MLE in mice significantly decreased the occurrence of pneumonia. Additionally, leaf powder of Melia (MLP) inactivated influenza viruses and viruses in the intestines of chickens. MLE and MLP may have applications as novel, safe biological disinfectants for use in humans and poultry.


Asunto(s)
Antivirales/administración & dosificación , Virus de la Influenza A/efectos de los fármacos , Virus de la Influenza A/crecimiento & desarrollo , Virus de la Influenza B/efectos de los fármacos , Virus de la Influenza B/crecimiento & desarrollo , Gripe Aviar/tratamiento farmacológico , Melia azedarach/química , Extractos Vegetales/administración & dosificación , Animales , Antivirales/química , Antivirales/aislamiento & purificación , Embrión de Pollo , Pollos , Femenino , Virus de la Influenza A/genética , Virus de la Influenza A/metabolismo , Virus de la Influenza B/genética , Virus de la Influenza B/metabolismo , Gripe Aviar/virología , Ratones , Extractos Vegetales/química , Extractos Vegetales/aislamiento & purificación , Hojas de la Planta/química , Enfermedades de las Aves de Corral/virología
5.
J Infect Dis ; 214(12): 1929-1936, 2016 Dec 15.
Artículo en Inglés | MEDLINE | ID: mdl-27923953

RESUMEN

BACKGROUND: In Indonesia, highly pathogenic avian influenza A(H5N1) virus has become endemic in poultry and has caused sporadic deadly infections in human. Since 2012, we have conducted fixed-point surveillance of avian influenza viruses at a live-poultry market in East Java, Indonesia. In this study, we examined the seroprevalence of avian influenza A(H5N1) virus infection among market workers. METHODS: Sera were collected from 101 workers in early 2014 and examined for antibody activity against avian A(H5N1) Eurasian lineage virus by a hemagglutination-inhibition (HI) assay. RESULTS: By the HI assay, 84% of the sera tested positive for antibody activity against the avian virus. Further analysis revealed that the average HI titer in 2014 was 2.9-fold higher than in 2012 and that seroconversion occurred in 44% of paired sera (11 of 25) between 2012 and 2014. A medical history survey was performed in 2016; responses to questionnaires indicated that none of workers had had severe acute respiratory illness during 2013. CONCLUSIONS: This study provides evidence of a high prevalence of avian A(H5N1) virus infection in 2013 among workers at a live-poultry market. However, because no instances of hospitalizations were reported, we can conclude the virus did not manifest any clinical symptoms in workers.


Asunto(s)
Crianza de Animales Domésticos , Anticuerpos Antivirales/sangre , Infecciones Asintomáticas/epidemiología , Subtipo H5N1 del Virus de la Influenza A/inmunología , Gripe Humana/epidemiología , Exposición Profesional , Animales , Pruebas de Inhibición de Hemaglutinación , Humanos , Indonesia/epidemiología , Aves de Corral , Estudios Seroepidemiológicos
6.
Cell Mol Life Sci ; 72(2): 357-66, 2015 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-25001578

RESUMEN

Influenza is a serious respiratory disease among immunocompromised individuals, such as the elderly, and its prevention is an urgent social issue. Influenza viruses rely on neuraminidase (NA) activity to release progeny viruses from infected cells and spreading the infection. NA is, therefore, an important target of anti-influenza drugs. A causal relationship between bacteria and influenza virus infection has not yet been established, however, a positive correlation between them has been reported. Thus, in this study, we examined the biological effects of oral mitis group streptococci, which are predominant constituents of human oral florae, on the release of influenza viruses. Among them, Streptococcus oralis ATCC 10557 and Streptococcus mitis ATCC 6249 were found to exhibit NA activity and their culture supernatants promoted the release of influenza virus and cell-to-cell spread of the infection. In addition, culture supernatants of these NA-producing oral bacteria increased viral M1 protein expression levels and cellular ERK activation. These effects were not observed with culture supernatants of Streptococcus sanguinis ATCC 10556 which lacks the ability to produce NA. Although the NA inhibitor zanamivir suppressed the release of progeny viruses from the infected cells, the viral release was restored upon the addition of culture supernatants of NA-producing S. oralis ATCC 10557 or S. mitis ATCC 6249. These findings suggest that an increase in the number of NA-producing oral bacteria could elevate the risk of and exacerbate the influenza infection, hampering the efficacy of viral NA inhibitor drugs.


Asunto(s)
Antivirales/farmacología , Gripe Humana/tratamiento farmacológico , Gripe Humana/microbiología , Neuraminidasa/metabolismo , Streptococcus mitis/enzimología , Streptococcus oralis/enzimología , Zanamivir/farmacología , Análisis de Varianza , Western Blotting , Técnica del Anticuerpo Fluorescente Indirecta , Humanos , Zanamivir/antagonistas & inhibidores
7.
Biol Pharm Bull ; 36(7): 1152-8, 2013.
Artículo en Inglés | MEDLINE | ID: mdl-23628892

RESUMEN

Pyrrole-imidazole (PI) polyamides are a novel group of gene-silencing compounds, which bind to a minor groove of double stranded (ds)DNA in a sequence-specific manner. To explore the RNA binding properties of PI polyamides targeting rat transforming growth factor-ß1 (TGF-ß1 Polyamide) and influenza A virus (PA polyamide), we designed dsRNAs with an identical sequence to the target DNA and analyzed RNA binding properties of the polyamide. Biacore assay showed fast binding of TGF-ß1 Polyamide to the dsRNA, whereas mismatch polyamide did not bind to the dsRNA. Dissociation equilibrium constant (KD) value was 6.7×10(-7) of the target dsRNA. These results indicate that PI polyamide could bind to RNA with a 2 log lower binding affinity than its DNA-binding affinity. We designed a PI polyamide targeting the panhandle stem region of influenza A virus. KD value of the PI polyamide to dsRNA targeting influenza A virus was 4.6×10(-7). Gel-shift assay showed that TGF-ß1 and PA polyamides bound to the appropriate dsDNA, whereas these PI polyamides did not show obvious gel-shift with the appropriate dsRNA. Structural modeling suggests that PI polyamide binds to the appropriate B-form dsDNA in the minor groove, whereas it does not fit in the minor groove to dsRNA. Thus PI polyamides have a lower binding affinity with target dsRNA than they do with dsDNA. The distinct binding properties of PI polyamides to dsRNA and dsDNA may be associated with differences of secondary structure and chemical binding properties between target RNA and DNA.


Asunto(s)
Silenciador del Gen/efectos de los fármacos , Imidazoles/farmacología , Virus de la Influenza A/química , Nylons/farmacología , Pirroles/farmacología , ARN Bicatenario/química , Factor de Crecimiento Transformador beta1/química , Animales , ADN/química , ADN/genética , Ensayo de Cambio de Movilidad Electroforética , Virus de la Influenza A/genética , Simulación del Acoplamiento Molecular , Conformación de Ácido Nucleico , ARN Bicatenario/genética , Ratas , Resonancia por Plasmón de Superficie , Factor de Crecimiento Transformador beta1/genética
8.
IJID Reg ; 8(Suppl): S1-S7, 2023 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-37799539

RESUMEN

Objectives: To determine the lineage distribution of the virus during the first wave of the pandemic in North Sumatra, Indonesia. Methods: A total of 20 samples with positive results based on reverse transcription-polymerase chain reaction were selected for virus culture and then performed whole-genome sequence analysis using next-generation sequencing which was applied by the Illumina MiSeq instrument. Results: Whole-genome sequence analysis revealed that the majority of our samples belong to lineages B.1.468 (n = 10), B.1 (n = 5), B.1.1 (n = 2), B.1.1.398 (n = 2), and B.6 (n = 1). Other unique amino acid mutations found in our samples were present in A58T on non-structural protein (NSP3) (70%), P323L on NSP12 (95%), Q57H on NS3 protein (75%), and D614G on S (100%). Conclusion: The SARS-CoV-2 lineage B.1.468 may be the main virus variant circulating in North Sumatra at the beginning of the emergence of COVID-19 cases in this province.

9.
PeerJ ; 11: e14917, 2023.
Artículo en Inglés | MEDLINE | ID: mdl-36846456

RESUMEN

Background: Influenza A viruses are a major pathogen that causes significant clinical and economic harm to many animals. In Indonesia, the highly pathogenic avian influenza (HPAI) H5N1 virus has been endemic in poultry since 2003 and has caused sporadic deadly infections in humans. The genetic bases that determine host range have not yet been fully elucidated. We analyzed the whole-genome sequence of a recent H5 isolate to reveal the evolution toward its mammalian adaptation. Methods: We determined the whole-genome sequence of A/chicken/East Java/Av1955/2022 (hereafter, "Av1955") from a healthy chicken in April 2022 and conducted phylogenetic and mutational analysis. Results: Phylogenetic analysis revealed that Av1955 belonged to the H5N1 clade 2.3.2.1c (Eurasian lineage). The six gene segments (PB1, PB2, HA, NP, NA, and NS) out of the eight segments derived from viruses of H5N1 Eurasian lineage, one (PB2) from the H3N6 subtype and the remaining one (M) from the H5N1 clade 2.1.3.2b (Indonesian lineage). The donor of the PB2 segment was a reassortant among three viruses of H5N1 Eurasian and Indonesian lineages and the H3N6 subtype. The HA amino acid sequence contained multiple basic amino acids at the cleavage site. Mutation analysis revealed that Av1955 possessed the maximal number of mammalian adaptation marker mutations. Conclusions: Av1955 was a virus of H5N1 Eurasian lineage. The HA protein contains an HPAI H5N1-type cleavage site sequence, while the virus was isolated from a healthy chicken suggesting its low pathogenicity nature. The virus has increased mammalian adaptation markers by mutation and intra- and inter-subtype reassortment, gathering gene segments possessing the most abundant maker mutations among previously circulating viruses. The increasing mammalian adaptation mutation in avian hosts suggests that they might be adaptive to infection in mammalian and avian hosts. It highlights the importance of genomic surveillance and adequate control measures for H5N1 infection in live poultry markets.


Asunto(s)
Subtipo H5N1 del Virus de la Influenza A , Virus de la Influenza A , Gripe Aviar , Humanos , Animales , Gripe Aviar/epidemiología , Pollos , Subtipo H5N1 del Virus de la Influenza A/genética , Indonesia , Filogenia , Virus de la Influenza A/genética , Aves de Corral , Mamíferos
10.
Analyst ; 137(15): 3520-7, 2012 Aug 07.
Artículo en Inglés | MEDLINE | ID: mdl-22705905

RESUMEN

The surfaces of silica-based sensor chips, designed for evanescent-field-coupled waveguide-mode sensors, were functionalized using various surface chemistries. The immobilization of molecular entities on the functionalized silica surfaces was monitored using various microscopic techniques (scanning electron, fluorescence, and atomic force microscopies). Further, gold nanoparticle-based signal enhancement analyses were performed with protein conjugation on different functionalized surfaces using a waveguide-mode sensor. Based on these analyses, the sensor surfaces modified with glutaraldehyde (Glu) and carbonyldiimidazole were found to be good for molecules of different sizes. In addition, it can be inferred that the Glu-modified surface may be suitable for small molecules with diameters around 5 nm owing to its surface roughness. The modified surface with carbonyldiimidazole is suitable for the direct immobilization of larger molecules especially for biomolecular assemblies without intermediate chemical modifications.


Asunto(s)
Glutaral/química , Imidazoles/química , Análisis por Matrices de Proteínas/métodos , Dióxido de Silicio/química , Estreptavidina/análisis , Microscopía Fluorescente , Tamaño de la Partícula , Análisis por Matrices de Proteínas/instrumentación , Propiedades de Superficie
11.
Sci Rep ; 12(1): 4835, 2022 03 22.
Artículo en Inglés | MEDLINE | ID: mdl-35318357

RESUMEN

To halt the pandemic of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), governments around the world have imposed policies, such as lockdowns, mandatory mask wearing, and social distancing. The application of disinfecting materials in shared public facilities can be an additional measure to control the spread of the virus. Copper is a prominent material with antibacterial and antiviral effects. In this study, we synthesized copper nanoparticles (CuNPs) as a surface coating agent and assessed their antiviral activity against SARS-CoV-2. CuNPs with a mean size of 254 nm in diameter were synthesized from copper sulfate as a source and were predominantly composed of copper oxide. The synthesized CuNPs were mixed with resin-based paint (CuNP/paint) and sprayed on the surface of stainless steel remnants. SARS-CoV-2 lost 97.8% infectivity on the CuNP/paint-coated surface after 30 min of exposure and more than 99.995% infectivity after 1 h of exposure. The inactivation rate was approximately 36-fold faster than that on the paint alone-coated and uncoated surfaces. The CuNP/paint-coated surface showed powerful inactivation of SARS-CoV-2 infectivity, although further study is needed to elucidate the inactivation mechanisms. Applications of CuNP/paint coatings to public or hospital facilities and other commonly touched areas are expected to be beneficial.


Asunto(s)
COVID-19 , Nanopartículas , Antivirales/farmacología , Antivirales/uso terapéutico , Control de Enfermedades Transmisibles , Cobre , Humanos , SARS-CoV-2
12.
Cytometry A ; 79(8): 653-60, 2011 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-21710641

RESUMEN

The initiation of translation in hepatitis C virus (HCV) occurs at the internal ribosome entry site (IRES) located at the 5'-end of its genomic RNA. To study the function of HCV IRES, we constructed a reporter plasmid that generates a bicistronic mRNA encoding two fluorescent proteins: cap-dependent DsRed2 and IRES-dependent Azami Green (AG). We introduced the plasmid into Huh7.5.1 and HEK293 cells and measured the relative IRES activity from the ratio of AG's signal to DsRed2's in individual cells using flow cytometry. To compare our method and a conventional biochemical method, we constructed a structurally similar reporter in which Renilla and Firefly luciferases replace DsRed2 and AG, respectively. With these systems, we found that the IRES A164G substitution decreased its activity, that interferon alpha affected the IRES activity in a cell type-specific manner, and that a synthetic micro-RNA targeting IRES was able to suppress the gene expression. In conclusion, the two methods were comparable in sensitivity in the studies of IRES mutations and host cell types. We discussed the significance of our findings and potential advantage of the cytometric assay: application to the molecular study of the HCV translation and to screening anti-IRES drugs.


Asunto(s)
Citometría de Flujo/métodos , Hepacivirus/metabolismo , Luciferasas de Luciérnaga/metabolismo , Luciferasas de Renilla/metabolismo , Biosíntesis de Proteínas , Ribosomas/metabolismo , Proteínas Virales/biosíntesis , Expresión Génica , Proteínas Fluorescentes Verdes , Células HEK293 , Hepacivirus/genética , Hepatocitos , Humanos , Interferón-alfa/metabolismo , Proteínas Luminiscentes , MicroARNs/genética , Ribosomas/genética
13.
Jpn J Infect Dis ; 73(5): 336-342, 2020 Sep 24.
Artículo en Inglés | MEDLINE | ID: mdl-32350224

RESUMEN

In Indonesia, the highly pathogenic avian influenza A/H5N1 virus has become endemic and has been linked with direct transmission to humans. From 2013 to 2014, we isolated avian influenza A/H5N1 and A/H3N6 viruses from poultry in Indonesia. This study aimed to reveal their pathogenicity in mammals using a mouse model. Three of the isolates, Av154 of A/H5N1 clade 2.3.2.1c, Av240 of A/H5N1 clade 2.1.3.2b, and Av39 of A/H3N6, were inoculated into BALB/c mice. To assess morbidity and mortality, we measured body weight daily and monitored survival for 20 d. Av154- and Av240-infected mice lost 25% of their starting body weight by day 7, while Av39-infected mice did not. Most of the Av154-infected mice died on day 8, while the majority of the Av240-infected mice survived until day 20. A 50% mouse lethal dose was calculated to be 2.0 × 101 50% egg infectious doses for Av154, 1.1 × 105 for Av240 and > 3.2 × 106 for Av39. The Av154 virus was highly virulent and lethal in mice without prior adaptation, suggesting its high pathogenic potential in mammals. The Av240 virus was highly virulent but modestly lethal, whereas the Av39 virus was neither virulent nor lethal. Several mammalian adaptive markers of amino acid residues were associated with the highly virulent and lethal phenotypes of the Av154 virus.


Asunto(s)
Subtipo H5N1 del Virus de la Influenza A/patogenicidad , Gripe Aviar/virología , Gripe Humana/virología , Aves de Corral/virología , Secuencia de Aminoácidos , Animales , Peso Corporal , Femenino , Humanos , Indonesia , Virus de la Influenza A/clasificación , Virus de la Influenza A/patogenicidad , Gripe Aviar/mortalidad , Gripe Humana/mortalidad , Ratones , Ratones Endogámicos BALB C , Modelos Animales , Infecciones por Orthomyxoviridae/mortalidad , Infecciones por Orthomyxoviridae/virología , Fenotipo , Virulencia
14.
J Virol Methods ; 156(1-2): 162-5, 2009 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-19027795

RESUMEN

The distribution of the matrix (M1) protein of influenza virus in infected cells was examined using immunostaining. The fixation method influenced strongly the immunofluorescence pattern of the M1 protein. The M1 protein was distributed uniformly in both the cytoplasm and in nuclei when cells that had been infected with virus were fixed with paraformaldehyde. In cells that had been fixed with methanol, however, nuclear dots of the M1 protein were clearly visible. The dots were evident at 8h post-inoculation. Up to 6h post-inoculation, only a diffuse distribution of the M1 protein was observed. The dots were co-localized with promyelocytic leukemia (PML) protein, a major component of nuclear domain 10 (ND10), also called PML oncogenic domains (PODs) or PML-nuclear bodies (NBs). These results indicate that the nuclear dots of the M1 protein in cells that had been fixed with methanol are not artifacts of the fixation method. Furthermore, methanol fixation is preferred for localization of the influenza M1 protein in nuclei using immunostaining.


Asunto(s)
Fijadores/química , Virus de la Influenza A/química , Proteínas de la Matriz Viral/análisis , Animales , Línea Celular , Técnica del Anticuerpo Fluorescente , Formaldehído/química , Metanol/química , Polímeros/química
15.
J Gastroenterol Hepatol ; 24(4): 599-604, 2009 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-19054262

RESUMEN

BACKGROUND/AIMS: Hepatitis E virus (HEV) in Bangladesh has not been adequately documented. We report HEV RNA and genotype detection in Bangladesh. METHODS: In total, 82 samples were used; 36 sporadic acute hepatitis (AH), 12 fulminant hepatitis (FH), 14 chronic liver disease (CLD) and 20 from an apparently healthy population (HP) positive for both immunoglobulin (Ig) M and IgG specific anti-HEV antibodies (anti-HEV). The male/female ratio was 61/21, ages 12-67 (mean 30.4) years. RNA was extracted, transcribed to cDNA and amplified in nt 6345-6490 (ORF2) of HEV. Nucleic and amino acid sequences were determined. Homology comparison between Bangladesh clones and other representative HEV clones and phylogenetic tree analyses were done. Relations between HEV RNA-positivity and clinical factors were analyzed. RESULTS: HEV RNA was positive in 9/36 (25.0%) of AH cases, 4/12 (33.3%) FH, 3/14 (21.4%) CLD and 0/20 (0%) HP samples; total 16/82 (19.5%). Four factors correlated significantly with HEV RNA-positivity (Mann-Whitney U test); alanine aminotransferase (ALT) (P = 0.0229), aspartate aminotransferase (AST) (P = 0.0448), and titers of IgG (P = 0.0208) and IgM (P = 0.0095) specific anti-HEV. The 16 HEV clones were divided mainly into two groups, A and B, including six different cDNA sub-groups. CONCLUSION: HEV RNA was found in sporadic AH and FH and sub-clinical CLD cases, but not in HP. HEV RNA-positivity was significantly related to values of ALT and AST and titers of IgG and IgM specific anti-HEV, with IgM specific anti-HEV showing the most significant relationship. All clones were genotype I, which is prevalent in South Asia.


Asunto(s)
Virus de la Hepatitis E/genética , Hepatitis E/diagnóstico , ARN Viral/sangre , Enfermedad Aguda , Adolescente , Adulto , Anciano , Secuencia de Aminoácidos , Bangladesh/epidemiología , Secuencia de Bases , Niño , Enfermedad Crónica , Femenino , Genotipo , Anticuerpos Antihepatitis/sangre , Hepatitis E/epidemiología , Hepatitis E/virología , Virus de la Hepatitis E/inmunología , Humanos , Inmunoglobulina G/sangre , Inmunoglobulina M/sangre , Masculino , Persona de Mediana Edad , Datos de Secuencia Molecular , Filogenia , Adulto Joven
16.
Microbiol Resour Announc ; 8(17)2019 Apr 25.
Artículo en Inglés | MEDLINE | ID: mdl-31023807

RESUMEN

We isolated an avian influenza A/H9N2 virus from an apparently healthy chicken at a live-poultry market in January 2018. This is the first report of a whole-genome sequence of A/H9N2 virus in Indonesia. Phylogenetic analyses indicated that intrasubtype reassortment of genome segments is involved in the genesis of the A/H9N2 virus.

17.
J Virol ; 81(23): 13259-64, 2007 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-17881441

RESUMEN

In a previous study, we demonstrated that humanized NOD/SCID/IL2Rgamma(null) (hNOG) mice constructed with human hematopoietic stem cells (HSCs) allow efficient human immunodeficiency virus type 1 (HIV-1) infection. However, HIV-1 infection could be monitored for only 43 days in the animals due to their short life spans. By transplanting HSCs without any myeloablation methods, the mice successfully survived longer than 300 days with stable engraftment of human cells. The mice showed high viremia state for more than the 3 months examined, with systemic HIV-1 infection and gradual decrease of CD4+ T cells analogous to that in humans. These capacities of the hNOG mice are very attractive for modeling mechanisms of AIDS progression and therapeutic strategy.


Asunto(s)
Modelos Animales de Enfermedad , Infecciones por VIH/virología , VIH-1/crecimiento & desarrollo , Trasplante de Células Madre Hematopoyéticas , Animales , Recuento de Linfocito CD4 , Femenino , Ratones , Ratones Noqueados , Ratones SCID , Viremia
18.
Virus Res ; 131(2): 170-9, 2008 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-17963943

RESUMEN

The Epstein-Barr virus (EBV)-encoded oncoprotein latent membrane protein 1 (LMP1) has an essential role in B-lymphocyte transformation by the virus and is expressed in certain EBV-associated tumors and lymphoproliferative disorders. By using the Flp-In/TREx-inducible expression system, we introduced LMP1 into two human cell lines, Jurkat and HEK-293, and found that in both of them the putative cellular oncogene Bcl-3 is rapidly induced following the expression of LMP1. Bcl-3 was also induced in Ramos cells after in vitro EBV infection and after transfection with an LMP1 expression vector. This LMP1-induced Bcl-3 expression is considered to be mediated by the transcription factor NF-kappaB, because (1) deletion of a critical NF-kappaB-binding site in the Bcl-3 promoter abolished its responsiveness to LMP1, (2) an IkappaB mutant that specifically inhibits NF-kappaB activity suppressed the LMP1-induced activation of the Bcl-3 promoter, and (3) an LMP1 mutant lacking its effector domain CTAR2, required for the activation of NF-kappaB, is severely impaired in its ability to induce Bcl-3. Western blot analyses showed that all EBV-infected and LMP1-expressing lymphoid cell lines express Bcl-3. These results suggest the possibility that Bcl-3 is involved in the pathogenesis of certain EBV-associated malignancies and lymphoproliferative disorders.


Asunto(s)
Herpesvirus Humano 4/fisiología , FN-kappa B/metabolismo , Proteínas Proto-Oncogénicas/biosíntesis , Factores de Transcripción/biosíntesis , Proteínas de la Matriz Viral/metabolismo , Proteínas del Linfoma 3 de Células B , Sitios de Unión , Western Blotting , Línea Celular , Genes Reporteros , Humanos , Luciferasas/genética , Luciferasas/metabolismo , FN-kappa B/antagonistas & inhibidores , Proteínas Proto-Oncogénicas/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Eliminación de Secuencia , Factores de Transcripción/genética , Proteínas de la Matriz Viral/genética
19.
Respirology ; 13(2): 203-14, 2008 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-18339017

RESUMEN

BACKGROUND AND OBJECTIVE: Airway epithelial cells, which are the initial site of influenza virus (IV) infection, participate in the inflammatory process through the expression of various genes. In this process, mitogen-activated protein kinase (MAPK) may be associated with the expression of many genes, but its precise role remains unknown. METHODS: A comprehensive analysis was performed of gene expression in human bronchial epithelial cells upon IV infection, using an Affymetrix gene chip containing 12 000 genes. Regulation of gene expression by MAPK was also analysed. RESULTS: A total of 5998 genes were detected. Upon IV infection, 165 genes were upregulated and 49 of these were interferon-stimulated genes. The functions of 129 genes, including 14 apoptosis-related genes and 6 antiviral genes, were well characterized; however, those of 36 genes were unknown. The expression of 29 genes was inhibited either by SB 203580, a specific inhibitor of p38 MAPK, or by CEP-11004, a specific inhibitor of the c-Jun-N-terminal kinase (JNK) cascade, and the percentage inhibition by SB 203580 correlated with that by CEP-11004, suggesting that p38 and JNK participate in a common downstream pathway involved in the regulation of gene expression. p38 MAPK- or JNK-dependent genes were functionally classified into diverse categories. CONCLUSIONS: Although further studies are needed to obtain a more complete understanding of gene expression and the role of MAPK in gene expression, the present results are important in understanding the molecular mechanisms involved in the response of bronchial epithelial cells to IV infection.


Asunto(s)
Bronquios/enzimología , Células Epiteliales/enzimología , Gripe Humana/enzimología , Proteínas Quinasas JNK Activadas por Mitógenos/fisiología , Proteínas Quinasas p38 Activadas por Mitógenos/fisiología , Bronquios/patología , Técnicas de Cultivo de Célula , Células Cultivadas , Humanos , Gripe Humana/patología , Análisis de Secuencia por Matrices de Oligonucleótidos , Regulación hacia Arriba/fisiología
20.
J Ethnopharmacol ; 118(1): 108-12, 2008 Jun 19.
Artículo en Inglés | MEDLINE | ID: mdl-18456441

RESUMEN

AIM OF THE STUDY: This investigation evaluated anti-influenza virus activity of 50% ethanol extract of the fruit of Chaenomeles sinensis K(OEHNE), which is widely used as a traditional Chinese medicine to treat throat diseases. MATERIAL AND METHODS: Type A and B influenza viruses were treated with the extract at various concentrations for 1h at room temperature; then the plaque titers of the treated viruses were determined. The neutralizing component in the extract was partially purified using HP20 column chromatography. RESULTS: Treatment with the extract at concentrations greater than 5mg/ml reduced the plaque titers of the both viruses to less than 10% of those of untreated viruses. The treatment inhibited viral hemagglutination activity, too. When the 50mg/ml extract was added to the culture medium after inoculation of the virus, viral NS2 protein synthesis was selectively inhibited and progeny virus was not detected in the infected cell medium. Partial purification showed that the neutralizing component consisted of high molecular weight polyphenols. CONCLUSION: High molecular weight polyphenols in the fruits of C. sinensis neutralizes influenza virus by inhibiting hemagglutination activity and by suppressing NS2 protein synthesis.


Asunto(s)
Antivirales/farmacología , Extractos Vegetales/farmacología , Rosaceae/química , Animales , Antivirales/administración & dosificación , Antivirales/aislamiento & purificación , Línea Celular , Perros , Relación Dosis-Respuesta a Droga , Flavonoides/administración & dosificación , Flavonoides/aislamiento & purificación , Flavonoides/farmacología , Frutas , Pruebas de Inhibición de Hemaglutinación , Virus de la Influenza A/efectos de los fármacos , Virus de la Influenza B/efectos de los fármacos , Riñón/efectos de los fármacos , Riñón/virología , Fenoles/administración & dosificación , Fenoles/aislamiento & purificación , Fenoles/farmacología , Extractos Vegetales/administración & dosificación , Extractos Vegetales/química , Polifenoles , Proteínas no Estructurales Virales/efectos de los fármacos , Proteínas no Estructurales Virales/metabolismo
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