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Porcine epidemic diarrhea virus (PEDV) is a highly pathogenic virus that can cause acute intestinal infectious diseases in both piglets and fattening pigs. The virus encodes at least 16 non-structural proteins, including nsp9, which has been shown to bind to single-stranded RNA. However, its function and mechanism remain unclear. In this study, we aimed to identify potential host proteins that interact with PEDV nsp9 using immunoprecipitation combined with mass spectrometry. The interactions were then confirmed by co-immunoprecipitation (Co-IP) and confocal laser scanning fluorescence techniques. The results showed that nsp9 interacts with HSPA8, Tollip, HSPA9 and TOMM70. Among them, overexpression of HSPA8 resulted in caused first upregulated and then down-regulated expression of nsp9, and promoted the proliferation of PEDV. Overexpression of Tollip significantly upregulated the expression of nsp9 and inhibited the proliferation of PEDV. Overexpression of TOMM70 significantly reduced the expression of nsp9, but did not show significant effect on the proliferation of PEDV. Overexpression of HSPA9 did not show significant effect on the expression of nsp9 and the proliferation of PEDV. These findings may facilitate further investigating the role of nsp9-interacting proteins in PEDV infection.
Subject(s)
Animals , Swine , Porcine epidemic diarrhea virus/genetics , Virus Replication , Proteins , Swine DiseasesABSTRACT
Rotavirus (RV) is one of the main pathogens causing diarrhea in children under five years old, but the mechanism of RV-infected diarrhea is still unclear. The RV genome encodes six structural proteins (VP1-VP4, VP6 and VP7) and six non-structural proteins (NSP1-NSP6), among which NSP4 can interact with other non-structural proteins or structural proteins of RV to produce corresponding biological functions, and is a key factor in the formation of RV morphology, the process of infection and the pathogenesis of diarrhea. In this paper, the current domestic and foreign studies on the structure and function of NSP4 are reviewed.
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@#Abstract：Objective To develop a colloidal gold immunochromatographic strip ⁃ based method for the rapid detection of Zika virus（ZIKV）NS1 antigen. Methods The gold nanoparticles modified with the anti⁃ZIKV NS1 monoclonal antibody as the detection probe were coated on the glass ⁃fiber pad. The anti ⁃ZIKV NS1 monoclonal antibody and the goat anti ⁃mouse polyclonal antibody were immobilized on a nitrocellulose membrane as the test line and the control line，respectively. In order to achieve critical results，the ratio of the optical density （OD）of the test line to that of the control line was compared. Serial diluted ZIKV NS1 standard antigen was applied to evaluate sensitivity of the immunoassay. The culture supernatant and serum samples for arboviruses（ZIKV，Dengue virus, Japanese encephalitis virus and Chikungunya virus） were utilized to demonstrate the specificity of the method. Results The detection result could read by naked eyes within 20 minutes. The visual cut ⁃off level for the test strip was achieved at 100 ng/mL of the Zika virus NS1 standard antigen. No cross⁃reactions with Dengue virus，Japanese encephalitis virus and Chikungunya virus were observed. The strip could remain good stability within 36 weeks whether stored in 4 ℃ or room temperature（22-25 ℃）. Conclusion Apart from stability， the method was convenient，rapid and specific for ZIKV NS1 antigen，which showed a promising potential in the point of care test and the screening test.
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Objective To investigate the molecular characteristics of immune response signaling molecules induced by transfection of coxsackievirus B2 ( CVB2 ) structural proteins into epithelial cells. Methods Recombinant eukaryotic expression plasmids containing the coding regions of CVB2 structural proteins VP1-VP4 were constructed and then transfected into 16HBE cells. Culture supernatants and cell ly-sates of the transfected 16HBE cells were collected. Expression of signaling molecules involved in innate im-mune responses in transfected 16HBE cells at mRNA level was detected by RT-Q-PCR. The proliferation of T cells co-cultured with culture supernatants and cell lysates of the transfected 16HBE cells was analyzed by ELISPOT. Results Expression of innate immunity-related signaling molecules such as TGF-β-activated ki-nase ( TAK) , NF-κB-inducing kinase ( NIK) , IκB kinase α ( IKKα) and IFN-β at mRNA level was up-regulated in 16HBE cells transfected with CVB2 structural proteins VP1-VP4. Both culture supernatants and cell lysates of the transfected 16HBE cells enhanced the proliferation of T cells. Conclusions CVB2 struc-tural proteins VP1-VP4 could enhance the expression of innate immunity-related signaling molecules to var-ying degrees and promote the activation of adaptive immunity.
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Urease decomposes urea to ammonia, and has application potential in agriculture and medical treatment. Urease proteins include structural proteins (UreA, UreB and UreC) and accessory proteins (UreD/UreH, UreE, UreF and UreG), each of them has its own unique role in urease maturation. The structural proteins form the active center of urease, and the accessory proteins are responsible for the delivery of nickel. We review here the structure and function of bacterial urease complexes, and how each protein interacts to complete the activation process. We hope to provide theoretical basis for the regulation of urease activity and the development of urease inhibitors.
Subject(s)
Bacterial Proteins , Nickel , Urease , MetabolismABSTRACT
Dengue virus (DENV) is one of the most significant mosquito-borne viral pathogens that spread in the tropical and subtropical areas causing severe diseases in humans such as dengue fever (DF), dengue hemorrhagic fever (DHF) and dengue shock syndrome (DSS). DENV RNA genome encodes three structural proteins and seven non-structural proteins. The non-structural protein 3 (NS3) contains two main functional domains: serine protease and RNA helicase. As a serine protease, NS3 together with host cell proteases can directly hydrolyze the ploy-protein that is translated from the viral genome into functional proteins. Functioning as a RNA helicase, it is closely related to the replication and transcriptional translation of the viral genome RNA. In this paper, the structure and functions of DENV NS3 and the research progress in related antiviral drugs were reviewed systematically.
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Bronchial asthma is a respiratory system disease char-acterized by airway remodeling as a pathological basis. Repeated inflammatory infiltration and tissue damage repair can lead to airway remodeling. At present,the mechanism of airway remod-eling is not comprehensive. Studies have shown that epithelial-mesenchymal transition (EMT) plays an important role in the genesis and development of airway remodeling. Airway epithelial cells can be induced to mesenchymal transition through a variety of secretion factors and signaling pathways,leading to airway re-modeling in asthma. This review summarizes the study of EMT and airway remodeling in asthma,providing a reference for clini-cal follow-up treatment and research.
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Resumen Introducción: El dengue perinatal es una patología de la que poco se sabe, los reportes disponibles describen riesgo de resultados perinatales adversos. Objetivo: Reportar un caso de dengue perinatal, como diagnóstico diferencial de sepsis neonatal, que debe tenerse en cuenta en zonas endémicas. Caso clínico: Recién nacido de una mujer de 23 años quien a las 36 semanas de gestación presentó cuadro de dengue con antígeno Non-Structural Protein 1 (NS1) positivo y anticuerpos anti-dengue negativos. Al sexto día de enfermedad dio a luz a un recién nacido sano, quien, al segundo día de vida, presentó fiebre sin otros hallazgos patológicos al examen físico, asociado a trombocitopenia severa (17.900 plaquetas/uL) y aumento de la proteína C reactiva, antígeno viral NS1 positivo e in-munoglobulina G (IgG) anti dengue positiva. Fue manejado con antibióticoterpia con ampicilina y gentamicina por protocolo de la institución para sepsis neonatal probable. El neonato mostró me joría clínica, con estabilidad hemodinámica y aumento significativo de plaquetas, siendo dado de alta. Conclusiones: El dengue en el embarazo trae consigo el riesgo de resultados perinatales adver sos, particularmente bajo peso al nacer y parto pre-término. Los hijos de madres diagnosticadas con dengue al final del embarazo deberían ser observados estrechamente con realización de hemograma seriado en los primeros días de vida, debido al riesgo de transmisión vertical.
Abstract Introduction: Few reports are available about perinatal dengue, with controversial results in regards the risk of perinatal outcome. Objective: To report a case of perinatal dengue as a differential diagno sis with neonatal sepsis, which must be considered in endemic areas. Clinical case: Male newborn of a 23 year-old female, who presented a Non-Structural Protein 1 (NS1) antigen positive to dengue at 36 weeks of gestation and negative anti-dengue antibodies. At day six of the illness a healthy newborn was born. On the second day of life the neonate presented fever with no other pathological findings on the physical exam, associated with severe thrombocytopenia (17,900 platelets/uL), increased C-reactive protein, a positive NS1 antigen, and positive anti-dengue immunoglobulin G (IgG). He was treated with ampicillin and gentamicin according the Institution protocol of neonatal sepsis. The newborn showed clinical improvement, with hemodynamic stability and significant increase of platelets, receiving the medical discharge. Conclusions: Dengue in pregnancy produces the risk of adverse perinatal outcomes, particularly low birth weight and preterm delivery. Children of mothers diagnosed with dengue at the end of pregnancy should be observed closely with serial hemograms during child's first days of life, due to the high risk of vertical transmission.
Subject(s)
Humans , Male , Female , Pregnancy , Infant, Newborn , Young Adult , Pregnancy Complications, Infectious/diagnosis , Infectious Disease Transmission, Vertical , Dengue/diagnosis , Dengue/transmission , Diagnosis, Differential , Neonatal Sepsis/diagnosisABSTRACT
The non-structural protein 1 is an important molecule of the viruses in flavivirus group including to Zika virus. Recently, the NS1 of Zika virus was discovered. There is still no complete information of the molecular interaction of NS1 of Zika virus which can be the clue for explanation for its pathogenesis and further drug search. Here the authors report the cleft analysis of NS1 of Zika virus and the result can be useful for future development of good diagnostic tool and antiviral drug finding for management of Zika virus.
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Objective:To prepare and preliminarily identify the monoclonal antibodies(mAbs) specifically against 3D protein of Enterovirus 71(EV71),using bioinformatics to predict the epitopes of 3D,with HBc protein as a carrier.Methods: Artificial screening of 3D protein epitope sequences by bioinformatic method,inserted into the major immunodominant region(MIR) area of Hepatitis B virus core protein(HBc),to construct the recombinant protein.BALB/c mice were immunized with the recombinant virus like particles(VLPs),to prepare the mAbs against 3D protein of EV71.Affinity chromatography technology was used to purify the mAb.The indirect ELISA,ELISPOT,immunofluorescence and immunohistochemistry staining methods were used to identify the characteristic of the mAb.Results: We displayed 3D(aa34-43),3D(aa61-76) and 3D(aa151-164) epitopes by constructing fusion protein using HBc VLPs as a vector,after hybridization,one positive hybridoma cell line(3E1) was selected by ELISA.The isotype of 3E1 was IgG2a.The results of immunofluorescence and immunohistochemistry staining assay showed that the mAb 3E1 could specifically recognize EV71.Conclusion: The prepared mAb 3E1 can specifically recognizes the EV71,which laid the foundation for the detection of virus and further study on 3D protein,and verified the bioinformatics technology combined with HBc carrier displaying peptides could prepare mAb quickly and efficiently.
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The non-structural protein 1 is an important molecule of the viruses in flavivirus group including to Zika virus. Recently, the NS1 of Zika virus was discovered. There is still no complete information of the molecular interaction of NS1 of Zika virus which can be the clue for explanation for its pathogenesis and further drug search. Here the authors report the cleft analysis of NS1 of Zika virus and the result can be useful for future development of good diagnostic tool and antiviral drug finding for management of Zika virus.
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Nonstructural protein 4 (NSP4) of Rotavirus has been designated as the first viral enterotoxin. Its role in viral replication is already well known. Intensive research over the past decade has shown the involvement of this protein in many cellular activities both in ‘expressed’ as well as ‘secreted’ form. It is responsible for increased intracellular calcium levels in the infected cell leading to a cascade of events which involve phospholipase C mediated secretion of Chloride ions. NSP4 also inhibits intestinal disaccharidases and sodium glucose symporter (SGLT1) so that complex sugars are retained resulting in malabsorption. It also alters actin in the villi resulting in their flattening and overall decreased absorptive area. NSP4 is associated with extracellular proteins and is hypothesized to have paracrine effects on neighbouring cells. Recent research has found it to be an activator of enteric nervous system too. All these factors contribute to the pathogenesis of diarrhoea which looks multifactorial and certainly very different from the bacterial toxin mediated diarrhoeas of E. coli and V. cholerae. We still don’t have the final word on this intriguing protein which is now a potential candidate for a vaccine against rotavirus. The aim of this review is to put forward the salient features of the research done to elucidate the functions of NSP4.
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Rotavirus is the most important pathogen causing severe diarrhea in infants and young children worldwide,and for rotavirus diarrhea there is no specific treatment so far.At present,there are a variety of claims on the pathogenesis of rotavirus infection,but none is conclusive.In the process of infection,rotavirus pathogens must overcome antiviral innate immune response that can replicate in host cell.Rotavirus encoded proteins play an important role in the process of rotavirus infection.This paperreviews the role of rotavirus encoding proteins in the pathogenesis of rotavirus infection.
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Dengue is known for its serious life-threatening complications. New rapid kits available recently in India target circulating non-structural protein (NS1) antigen from day one onwards. The sensitivity and specifi city of a newly introduced rapid combo kit against two conventional ELISA kits is assessed. The performance of this kit is quite satisfactory since excellent agreement of 94.26% was observed with particular reference to NS1 antigen detection among all three kits namely Rapid SD Bioline dengue Duo (SD Korea), InBios DENV Detect NS1 ELISA, USA and dengue Early ELISA, Panbio, Australia. The false positivity of the rapid kit is very low since its specifi city as for as NS1 antigen detection is concerned is 98.33%. The use of combination kit helps to detect additional cases of dengue, which are negative for NS1 antigen but positive for IgM and/or IgG antibodies, thus facilitating early diagnosis in remote areas and small laboratories.
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Aim To investigate the function of fenofi-brate on PAN ( puromycin aminonucleoside )-induced podocyte injury. Methods SD female rats of 18-week-old were randomly assigned into 3 groups ( n =6 ) . Mice in PAN group and fenofibrate treated group received a single intravenous injection of PAN ( 65 mg ·kg-1 ) , while those in control group received equal volume of saline. Mice in fenofibrate treated group re-ceived 40 mg · kg-1 · d-1 of fenofibrate ( intragastric administration ) on day 1 after PAN injection , while those in PAN group and control group received equal volume of vehicle. 24 hours urine samples from all group were collected on day 0(1 day before PAN injec-tion), day 6, day 10. The 24 hours urine protein was detected by Bradford assay. All the rats were sacrificed 10 days after the induction of podocyte injury, and glo-merulus sample were collected. The expression of podocyte injury marker and transcription level in apop-tosis, podocyte cytoskeleton protein, slit diaphragm protein were evaluated by Western blot and real-time PCR. Results Compared with the control group, 10 days after injection of PAN, 24 hours urine protein was obviously increased, and the expression and transcrip-tion level of podocyte injury marker desmin, apoptosis, podocyte cytoskeleton protein, slit diaphragm protein were upregulated greatly, however, those were signifi-cantly lower in fenofibrate treated group as compared with those in PAN group. Conclusions PPAR-α ago-nist fenofibrate can ameliorate PAN-induced glomerulus podocyte injury, and the mechanism involved may be associated with inhibition of the mitochondria apopto-sis, TGF-β/Smad pathway and p38 pathway.
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In the present study, 89 porcine reproductive and respiratory syndrome virus (PRRSV) isolates in China during 2007 to 2012 were randomly selected from the GenBank genetic sequence database. Evolutionary characteristics of these isolates were analyzed based on the sequences of non-struc-tural protein 2 (Nsp2) and glycoprotein 5 (GP5). The genetic variations of the isolates were also compared with six representative strains. The results showed that a high degree of genetic diversity exists among the PRRSV population in China. Highly pathogenic PRRSV isolates, with a discon-tinuous deletion of a 30 amino acid residue in the Nsp2 region, remained the most dominant virus throughout 2007-2012 in China. Owing to the extensive use of representative vaccine strains, natu-ral recombination events occurred between strains. Three isolates-HH08, DY, and YN-2011-were more closely related to vaccine strains than the other isolates. Both YN-2011 and DY were the evolu-tionary products of recombination events between strains SP and CH-1R. The results of the present study provide useful information for the epidemiology of PRRSV as well as for vaccine development.
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Out of 200 serum samples collected from cattle (142) and buffaloes (58) of various ages and sexand subjected to latex agglutination test (LAT) using serotype specific peptides (O, A, Asia 1) and also with peptide for non-structural protein 2B (NSP-2B), 114 (70%) samples were positive against FMDV type ‘O’, 102 (51%) against serotype ‘A’ and 104 (52%) against serotype ‘Asia 1’. With NSP-2B peptide a total of 71 (35.5%) samples were positive. The results suggest that LAT could be used for the diagnosis of foot and mouth disease virus as it is easy, cheap and effective test.
Subject(s)
Amino Acid Sequence , Animals , Cattle , Foot-and-Mouth Disease/immunology , Foot-and-Mouth Disease Virus/classification , Latex Fixation Tests/methods , Microspheres , Molecular Sequence Data , Peptides/chemistry , Peptides/immunology , Serotyping , Vaccination , Viral Nonstructural Proteins/immunologyABSTRACT
Parvoviruses are small, 260-Å-diameter, icosahedral, non-enveloped, single-stranded DNA viruses with a genome of approximately 5 kb. Non structural protein, (NS-1) is especially relevant, being both essential for virus replication and the main factor responsible for virus pathogenicity and cytotoxicity. This protein has also been reported to possess the property of killing of transformed cells. The present study was carried out to clone, characterize and express the NS-1 gene of canine parvovirus. NS-1 complete CDS 2020bp was amplified, cloned into eukaryotic expression vector pcDNA 3.1(+), sequenced and characterized by in vitro expression analysis. Functional activity of recombinant construct, pcDNA.cpv.NS-1, was evaluated by RT-PCR and flow cytometry for the expression of NS-1 specific mRNA and NS-1 protein, respectively, in transfected HeLa cells. This recombinant plasmid may serve as an important tool to evaluate the apoptotic potential of NS-1 protein of canine parvovirus in cultured HeLa cells.
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The A Iran 05 foot-and-mouth disease virus (FMDV) subtype was detected in Iran during 2005 and has proven to be highly virulent. This study was undertaken to focus on molecular and phylogenetic analysis of 3A and 3B coding-regions in the A Iran 05 field isolate. To assess the genetic relatedness of A Iran 05 isolate the nucleotide and predicted amino acid sequences of the 3AB region of type A FMDV isolates were compared with twenty previously described type A FMDV isolates. The phylogenetic tree based on the 672 bp 3AB gene sequences of type A FMDV from thirteen different locations clustered them into five distinct lineages. The A Iran 05 isolate clustered in lineage A along with four type A variants and was closely matched with viruses isolated in Turkey and Pakistan during 2005~2006. The number of protein sequence differences exhibited by each of the isolates revealed that A Iran 05 isolate contains three amino acid substitutions at positions 47 and 119 of 3A and 27 of the 3B coding region. The nucleotide identity between A Iran 05 and the other four isolates of lineage A was estimated to be 98%.
Subject(s)
Amino Acid Sequence , Base Sequence , Cluster Analysis , DNA Primers/genetics , Foot-and-Mouth Disease Virus/genetics , Iran , Molecular Sequence Data , Phylogeny , Reverse Transcriptase Polymerase Chain Reaction , Sequence Alignment , Sequence Analysis, DNA , Sequence Homology , Species Specificity , Viral Nonstructural Proteins/geneticsABSTRACT
Viruses including baculoviruses are obligatory parasites, as their genomes do not encode all the proteins required for replication. Therefore, viruses have evolved to exploit the behavior and the physiology of their hosts and often eoevolved with their hosts over millions of years. Recent comparative analyses of complete genome sequences of baculoviruses revealed the patterns of gene acquisitions and losses that have occurred during baculovirus evolution. In addition, knowledge of virus genes has also provided understanding of the mechanism of baculovirus infection including replication, species-specific virulence and host range. The Bm8 gene of Bombyx mori nucleopolyhedrovirus (NPV) and its homologues are found only in group I NPV genomes. The Autographa californica NPV Acl6 gene is a homologue of Bm8 and, encodes a viral structural protein. It has been shown that Bm8/Ac 16 interacts with baculoviral and cellular proteins. Bm8/Ac 16 interacts with baculoviral IE1 that is facilitated by coiled coil domains, and the interaction with IE1 is important for Bin8 function. Ac16 also forms a complex with viral FP25 and cellular actin and associates with membranes via palmitoylation. These data suggested that this gene family encodes a multifunctional protein that accomplishes specific needs of group INPVs.