280 research outputs found

    Genomska tipizacija i filogenetska analiza izolata pasjeg parvovirusa izdvojenih u državi Odisha u Indiji.

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    Canine parvovirus type 2 (CPV-2) comprises three major antigenic variants CPV-2a, CPV-2b and CPV-2c. Their mutated variants in geographically distinct locations need to be investigated to understand viral evolution and for development of effective management measures. In the present study, 71 faecal and 12 blood samples from suspected dogs in the state of Odisha, India were analyzed by PCR. Faecal lysate, extracted by the fast boiling method was found to be more sensitive as a template for PCR compared to DNA extracted from faecal samples by the phenol-chloroform method. The results revealed 29 positive cases (583 bp amplicon) out of 71 faecal samples, and 5 positive cases out of 12 blood samples examined, with a few variations in the results from blood and faecal samples in the same cases, thus suggesting the necessity of screening both blood and faecal samples for diagnosis. Restriction digestion of the 583 bp PCR amplicon with MboII (PCR-RFLP) confirmed the strain not to be CPV-2c. Further sequencing of the 583 bp fragments recognized the variant as one of the mutated CPV-2a strain. Interestingly, an additional presence of CPV-2a mutant of 525 bp was observed in eleven of the positive faecal samples, along with the 583 bp fragment in PCR that needs further characterization. These two CPV-2a variants shared a common clade with other CPV-2a variants in the phylogenetic tree separating CPV-2b and CPV-2c. Our results confirm the dynamic changes in CPV variants and emphasize the importance of CPV surveillance for understanding of viral epidemiology.Pasji parvovirus tip 2 (PPV-2) ima tri glavne antigenske varijante: PPV-2a, PPV-2b i PPV-2c. Radi razumijevanja njegove evolucije i razvijanja učinkovitih mjera suzbijanja potrebno je istražiti njegove mutante iz različitih geografskih područja. U ovom je radu lančanom reakcijom polimerazom bio pretražen 71 uzorak fecesa i 12 uzoraka krvi pasa sa sumnjom na parvovirusnu infekciju u državi Odisha u Indiji. Postupak dobivanja fekalnog lizata brzim ključanjem pri 100 °C pokazao se osjetljivijim u odnosu na ekstrakciju DNA iz uzoraka fecesa fenol-kloroformom. Rezultati su pokazali da je od 71 pretraženog uzorka fecesa 29 bilo pozitivnih (583 bp umnožak), a od 12 pretraženih uzoraka krvi sedam pozitivnih s različitim nalazima kod istih slučajeva, što govori da je za postavljanje dijagnoze potrebno pretražiti oba uzorka od iste životinje. Cijepanje odsječka 583 bp restrikcijskim enzimom MboII (PCR-RFLP) pokazalo je da izdvojeni soj ne pripada PPV-2c. Daljnjim sekvencioniranjem fragmenata od 583 bp pokazalo se da izolat pripada mutiranoj varijanti PPV-2a. Zanimljivo je da je u 11 pozitivnih uzoraka fecesa usporedno s fragmentima od 583 bp bila dokazana i prisutnost mutanta PPV-2a s fragmentom od 525 bp što iziskuje daljnju karakterizaciju. Te dvije varijante PPV-2a svrstane su u zajedničku skupinu u filogenetskom stablu, različitu od PPV-2b i PPV-2c. Naši rezultati potvrđuju dinamiku promjena varijanata PPV s naglaskom na važnost istraživanja za razumijevanje njegove epizootiologije

    Comparative and temporal transcriptome analysis of peste des petits ruminants virus infected goat peripheral blood mononuclear cells

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    Peste des petits ruminanats virus (PPRV), a morbillivirus causes an acute, highly contagious disease – peste des petits ruminants (PPR), affecting goats and sheep. Sungri/96 vaccine strain is widely used for mass vaccination programs in India against PPR and is considered the most potent vaccine providing long-term immunity. However, occurrence of outbreaks due to emerging PPR viruses may be a challenge. In this study, the temporal dynamics of immune response in goat peripheral blood mononuclear cells (PBMCs) infected with Sungri/96 vaccine virus was investigated by transcriptome analysis. Infected goat PBMCs at 48 h and 120 h post infection revealed 2540 and 2000 differentially expressed genes (DEGs), respectively, on comparison with respective controls. Comparison of the infected samples revealed 1416 DEGs to be altered across time points. Functional analysis of DEGs reflected enrichment of TLR signaling pathways, innate immune response, inflammatory response, positive regulation of signal transduction and cytokine production. The upregulation of innate immune genes during early phase (between 2-5 days) viz. interferon regulatory factors (IRFs), tripartite motifs (TRIM) and several interferon stimulated genes (ISGs) in infected PBMCs and interactome analysis indicated induction of broad-spectrum anti-viral state. Several Transcription factors – IRF3, FOXO3 and SP1 that govern immune regulatory pathways were identified to co-regulate the DEGs. The results from this study, highlighted the involvement of both innate and adaptive immune systems with the enrichment of complement cascade observed at 120 h p.i., suggestive of a link between innate and adaptive immune response. Based on the transcriptome analysis and qRT-PCR validation, an in vitro mechanism for the induction of ISGs by IRFs in an interferon independent manner to trigger a robust immune response was predicted in PPRV infection

    Modulation of Host miRNAs Transcriptome in Lung and Spleen of Peste des Petits Ruminants Virus Infected Sheep and Goats

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    Peste des petits ruminants (PPR) is one of the highly contagious viral disease, characterized by fever, sore mouth, conjunctivitis, gastroenteritis, and pneumonia, primarily affecting sheep and goats. Reports suggested variable host response in goats and sheep and this host response vis-a-vis the expression of microRNAs (miRNAs) has not been investigated. Here, miRNAs were sequenced and proteomics data were generated to identify the role of differentially expressed miRNA (DEmiRNA) in PPR virus (PPRV) infected lung and spleen tissues of sheep and goats. In lungs, 67 and 37 DEmiRNAs have been identified in goats and sheep, respectively. Similarly, in spleen, 50 and 56 DEmiRNAs were identified in goats and sheep, respectively. A total of 20 and 11 miRNAs were found to be common differentially expressed in both the species in PPRV infected spleen and lung, respectively. Six DEmiRNAs—miR-21-3p, miR-1246, miR-27a-5p, miR-760-3p, miR-320a, and miR-363 were selected based on their role in viral infections, apoptosis, and fold change. The target prediction analysis of these six selected DEmiRNAs from the proteome data generated, revealed involvement of more number of genes in lung and spleen of goats than in sheep. On gene ontology analysis of host target genes these DEmiRNAs were found to regulate several immune response signaling pathways. It was observed that the pathways viz. T cell receptor signaling, Rap1 signaling, Toll-like receptor signaling, and B cell receptor signaling governed by DEmiRNAs were more perturbed in goats than in sheep. The data suggests that PPRV-induced miR-21-3p, miR-320a, and miR-363 might act cooperatively to enhance viral pathogenesis in the lung and spleen of sheep by downregulating several immune response genes. The study gives an important insight into the molecular pathogenesis of PPR by identifying that the PPRV—Izatnagar/94 isolate elicits a strong host response in goats than in sheep

    Dysregulated miRNAome and Proteome of PPRV Infected Goat PBMCs Reveal a Coordinated Immune Response

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    In this study, the miRNAome and proteome of virulent Peste des petits ruminants virus (PPRV) infected goat peripheral blood mononuclear cells (PBMCs) were analyzed. The identified differentially expressed miRNAs (DEmiRNAs) were found to govern genes that modulate immune response based on the proteome data. The top 10 significantly enriched immune response processes were found to be governed by 98 genes. The top 10 DEmiRNAs governing these 98 genes were identified based on the number of genes governed by them. Out of these 10 DEmiRNAs, 7 were upregulated, and 3 were downregulated. These include miR-664, miR-2311, miR-2897, miR-484, miR-2440, miR-3533, miR-574, miR-210, miR-21-5p, and miR-30. miR-664 and miR-484 with proviral and antiviral activities, respectively, were upregulated in PPRV infected PBMCs. miR-210 that inhibits apoptosis was downregulated. miR-21-5p that decreases the sensitivity of cells to the antiviral activity of IFNs and miR-30b that inhibits antigen processing and presentation by primary macrophages were downregulated, indicative of a strong host response to PPRV infection. miR-21-5p was found to be inhibited on IPA upstream regulatory analysis of RNA-sequencing data. This miRNA that was also highly downregulated and was found to govern 16 immune response genes in the proteome data was selected for functional validation vis-a-vis TGFBR2 (TGF-beta receptor type-2). TGFBR2 that regulates cell differentiation and is involved in several immune response pathways was found to be governed by most of the identified immune modulating DEmiRNAs. The decreased luciferase activity in Dual Luciferase Reporter Assay indicated specific binding of miR-21-5p and miR-484 to their target thus establishing specific binding of the miRNAs to their targets.This is the first report on the miRNAome and proteome of virulent PPRV infected goat PBMCs

    Centrality evolution of the charged-particle pseudorapidity density over a broad pseudorapidity range in Pb-Pb collisions at root s(NN)=2.76TeV

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    Towards Understanding of Artificial Intelligence in Accounting Profession

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    Accountants have embraced the emission of automation over many years to get better the efficiency and effectiveness of their work. But technology has not been able to replace the need for expert knowledge and decision-making. Earlier generations of ‘intelligent systems have usually demonstrated the progressing power of human expertise and the restrictions of machines. In the upcoming decades, intelligent systems must take over more and better decision-making tasks from humans. While accountant has been using technology for a lot of years to improve what they do and deliver more value to businesses, this is an opportunity to reimagine and radically improve the quality of business and investment decisions which is the ultimate purpose of the profession. Accountants, as expert decision-makers, use both ways of thinking they apply their knowledge to specific situations to make reasoned decisions, although also make quick intuitive decisions based on extensive experience in their field. Today, AI is being used for image recognition, object identification, detection, classification, and automated geophysical feature detection. These are underlying tasks that once required the input of a human. Focusing on how artificial intelligence will impact accountants, AI will very soon help the organization to automate much of the routine and repetitive activities that are undertaken on a daily, weekly or annual basis. It will also help the organization to empower quick decision-making to create smart insights examine huge quantities of data with ease

    Observational constraint on interacting Tsallis holographic dark energy in logarithmic Brans–Dicke theory

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    In this paper, we investigate the dark energy phenomenon by studying the Tsallis holographic dark energy within the framework of Brans–Dicke (BD) scalar–tensor theory of gravity (Brans and Dicke in Phys. Rev. 124:925, 1961). In this context, we choose the BD scalar field ϕ\phi as a logarithmic function of the average scale factor a(t) and Hubble horizon as the IR cutoff (L=H1L=H^{-1}). We reconstruct two cases of non-interacting and interacting fluid (dark sectors of cosmos) scenario. The physical behavior of the models are discussed with the help of graphical representation to explore the accelerated expansion of the universe. Moreover, the stability of the models are checked through squared sound speed vs2v_s^2. The well-known cosmological plane i.e., ωdeωde\omega _{de}-\omega ^{\prime }_{de} is constructed for our models. We also include comparison of our findings of these dynamical parameters with observational constraints. It is also quite interesting to mention here that the results of deceleration, equation of state parameters and ωdeωde\omega _{de}-\omega ^{\prime }_{de} plane coincide with the modern observational data

    Genomska tipizacija i filogenetska analiza izolata pasjeg parvovirusa izdvojenih u državi Odisha u Indiji.

    Get PDF
    Canine parvovirus type 2 (CPV-2) comprises three major antigenic variants CPV-2a, CPV-2b and CPV-2c. Their mutated variants in geographically distinct locations need to be investigated to understand viral evolution and for development of effective management measures. In the present study, 71 faecal and 12 blood samples from suspected dogs in the state of Odisha, India were analyzed by PCR. Faecal lysate, extracted by the fast boiling method was found to be more sensitive as a template for PCR compared to DNA extracted from faecal samples by the phenol-chloroform method. The results revealed 29 positive cases (583 bp amplicon) out of 71 faecal samples, and 5 positive cases out of 12 blood samples examined, with a few variations in the results from blood and faecal samples in the same cases, thus suggesting the necessity of screening both blood and faecal samples for diagnosis. Restriction digestion of the 583 bp PCR amplicon with MboII (PCR-RFLP) confirmed the strain not to be CPV-2c. Further sequencing of the 583 bp fragments recognized the variant as one of the mutated CPV-2a strain. Interestingly, an additional presence of CPV-2a mutant of 525 bp was observed in eleven of the positive faecal samples, along with the 583 bp fragment in PCR that needs further characterization. These two CPV-2a variants shared a common clade with other CPV-2a variants in the phylogenetic tree separating CPV-2b and CPV-2c. Our results confirm the dynamic changes in CPV variants and emphasize the importance of CPV surveillance for understanding of viral epidemiology.Pasji parvovirus tip 2 (PPV-2) ima tri glavne antigenske varijante: PPV-2a, PPV-2b i PPV-2c. Radi razumijevanja njegove evolucije i razvijanja učinkovitih mjera suzbijanja potrebno je istražiti njegove mutante iz različitih geografskih područja. U ovom je radu lančanom reakcijom polimerazom bio pretražen 71 uzorak fecesa i 12 uzoraka krvi pasa sa sumnjom na parvovirusnu infekciju u državi Odisha u Indiji. Postupak dobivanja fekalnog lizata brzim ključanjem pri 100 °C pokazao se osjetljivijim u odnosu na ekstrakciju DNA iz uzoraka fecesa fenol-kloroformom. Rezultati su pokazali da je od 71 pretraženog uzorka fecesa 29 bilo pozitivnih (583 bp umnožak), a od 12 pretraženih uzoraka krvi sedam pozitivnih s različitim nalazima kod istih slučajeva, što govori da je za postavljanje dijagnoze potrebno pretražiti oba uzorka od iste životinje. Cijepanje odsječka 583 bp restrikcijskim enzimom MboII (PCR-RFLP) pokazalo je da izdvojeni soj ne pripada PPV-2c. Daljnjim sekvencioniranjem fragmenata od 583 bp pokazalo se da izolat pripada mutiranoj varijanti PPV-2a. Zanimljivo je da je u 11 pozitivnih uzoraka fecesa usporedno s fragmentima od 583 bp bila dokazana i prisutnost mutanta PPV-2a s fragmentom od 525 bp što iziskuje daljnju karakterizaciju. Te dvije varijante PPV-2a svrstane su u zajedničku skupinu u filogenetskom stablu, različitu od PPV-2b i PPV-2c. Naši rezultati potvrđuju dinamiku promjena varijanata PPV s naglaskom na važnost istraživanja za razumijevanje njegove epizootiologije

    Growth of textured nanocrystalline cobalt ferrite thin films by pulsed laser deposition

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    Cobalt ferrite thin films have been deposited on fused quartz substrates by pulsed laser deposition at various substrate temperatures, T-S (25 degrees C, 300 degrees C, 550 degrees C and 750 degrees C). Single phase, nanocrystalline, spinel cobalt ferrite formation is confirmed by X-ray diffraction (XRD) for T-S >= 300 degrees C. Conventional XRD studies reveal strong (111) texturing in the as deposited films with T-S >= 550 degrees C. Bulk texture measurements using X-ray orientation distribution function confirmed (111) preferred orientation in the films with T-S >= 550 degrees C. Grain size (13-16 nm for T-S >= 300 degrees C) estimation using grazing incidence X-ray line broadening analysis shows insignificant grain growth with increasing T-S, which is in good agreement with grain size data obtained from transmission electron microscopy

    <i><span style="font-size:11.0pt;font-family: "Times New Roman";mso-fareast-font-family:"Times New Roman";mso-bidi-font-family: Mangal;mso-ansi-language:EN-GB;mso-fareast-language:EN-US;mso-bidi-language: HI" lang="EN-GB">In vitro</span></i><span style="font-size:11.0pt;font-family: "Times New Roman";mso-fareast-font-family:"Times New Roman";mso-bidi-font-family: Mangal;mso-ansi-language:EN-GB;mso-fareast-language:EN-US;mso-bidi-language: HI" lang="EN-GB"> cloning of canine parvovirus NS1 gene and reporter gene GFP in eukaryotic expression vector pVIVO2-mcs and characterization of the double gene construct in mammalian cells</span>

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    41-46The use of chemotherapy and/or radiotherapy for treatment of cancer is limited due to genotoxic side effects on healthy cells, involvement of anti-apoptotic signal transduction pathways that prevent cell death, and requirement of functional p53 for induction of apoptosis in cancerous cells. Efforts are beings made worldwide to develop new anticancer therapies as an alternative to chemotherapy. And viral gene therapy is one of the most potent therapeutics that is being ventured worldwide. Canine parvovirus-2 (CPV-2) is one of those viruses that have an inherent oncolytic property. The non-structural protein-1 (NS1 protein) of CPV-2 plays a major role in parvoviral cytotoxicity and pathogenicity in permissive cells. The oncolytic potential of CPV2-NS1 has been established in vitro. Prior to taking up the in vivo studies, the present study was undertaken to clone Canine Parvovirus NS1 gene and reporter gene GFP in eukaryotic expression vector pVIVO2-mcs, and to characterize the double construct in mammalian cells. The genes were successfully cloned in pVIVO2-mcs and characterized for their expression as demonstrated by fluorescence microscopy and immunofluorescence staining. This characterized double gene construct will further be used to evaluate the oncolytic potential of CPV-2 NS1 in experimentally induced in vivo tumour model
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