Production of Peste des Petits Ruminants (PPR) Vaccine in Tide Motion Bioreactor Using Vero Cells

2021 ◽  
pp. 15-21
Author(s):  
Amitha Reena Gomes ◽  
R. Apsana ◽  
J. V. Savitha ◽  
M. Sobha Rani ◽  
S. M. Byregowda ◽  
...  
Keyword(s):  
Vero Cells ◽  
Peste Des Petits Ruminants

scholarly journals Recovery and Characterization of a Chimeric Rinderpest Virus with the Glycoproteins of Peste-des-Petits-Ruminants Virus: Homologous F and H Proteins Are Required for Virus Viability

2000 ◽  
Vol 74 (19) ◽  
pp. 9039-9047 ◽  
Author(s):  
Subash C. Das ◽  
Michael D. Baron ◽  
Thomas Barrett
Keyword(s):  
Clinical Signs ◽  
Vero Cells ◽  
Virus Genome ◽  
Peste Des Petits Ruminants ◽  
Live Virus ◽  
Domestic Ruminants ◽  
Rescue System ◽  
Ppr Virus ◽  
Rescue Procedure

ABSTRACT Rinderpest (RP) and peste-des-petits-ruminants (PPR) are two important diseases of domestic ruminants. To improve on currently available vaccines against PPR, we have created cDNA copies of the RP virus genome in which either the fusion (F) or hemagglutinin (H) gene, or both, was replaced with the corresponding gene from PPR virus. It was necessary to develop a modified rescue system in which the T7 RNA polymerase was provided by a recombinant fowlpox virus and the entire rescue procedure took place in Vero cells before we could obtain live virus from these chimeric constructs. No virus was recovered when only one of the glycoprotein genes was changed, but a chimeric virus containing both F and H genes from PPR virus was reproducibly rescued from cDNA, indicating that a virus-specific functional interaction takes place between the F and H proteins. The rescued virus expressing the PPR glycoproteins grew more slowly in tissue culture than either parental virus and formed abnormally large syncytia. Goats infected with the chimera showed no adverse reaction, as assessed by clinical signs, temperature, leukocyte count, virus isolation, and serology, and were protected from subsequent challenge with wild-type PPR virus.

scholarly journals Antiviral Effectivity of Favipiravir Against Peste Des Petits Ruminants Virus Is Mediated by the JAK/STAT and PI3K/AKT Pathways

2021 ◽  
Vol 8 ◽  
Author(s):  
Weifeng Zhang ◽  
Hualong Deng ◽  
Yanfen Liu ◽  
Shaohong Chen ◽  
You Liu ◽  
...  
Keyword(s):  
Western Blotting ◽  
Effective Means ◽  
Small Ruminants ◽  
Antiviral Agent ◽  
Vero Cells ◽  
Host Cells ◽  
Peste Des Petits Ruminants ◽  
Expression Levels ◽  
Qrt Pcr

Peste des petits ruminants virus (PPRV), belonging to the genus Morbillivirus in the family Paramyxoviridae, causes severe infectious disease in small ruminants and has been rapidly spreading in many parts of Africa, the Middle East, and Asia. Although vaccination is considered to be an effective means of controlling PPR, the heat-sensitive nature of the vaccines against PPRV greatly limits their application in areas with a hot climate. In the present study, we investigated the anti-PPRV effects of favipiravir and sought to identify the underlying mechanisms in vitro using the Vero cell line. MTT assays, Western blotting, indirect immunofluorescence assays, virus plaque formation assays, and qRT-PCR were used to assess the effects of favipiravir on the life cycle of PPRV and the expression of RNA-dependent RNA polymerase (RdRp). Additionally, the expression levels of JAK1, STAT1, phosphorylated (p)-STAT1, PI3K, AKT, and p-AKT, as well as those of signaling molecules acting downstream of the JAK/STAT and PI3K/AKT signaling pathways, were determined by Western blotting and qRT-PCR. The results indicated that, in PPRV-infected, favipiravir-treated Vero cells, the attachment, invasion, replication, and release of PPRV were significantly inhibited, as was the expression of RdRp, when compared with that in untreated PPRV-infected cells. Furthermore, in favipiravir-treated cells, the expression of JAK1 and STAT1 was downregulated, whereas that of p-STAT1 was significantly upregulated. Similarly, the expression levels of PKR, IRF9, ISG54, and MxA proteins that are associated with innate antiviral activity in host cells were also markedly increased. Moreover, with favipiravir treatment, the expression of PI3K and p-AKT and the p-AKT/AKT ratio were significantly decreased, whereas the expression of AKT was noticeably upregulated. The expression of GSK3, NF-κB p65, p-NF-κB p65, and BAD was also increased with favipiravir treatment, while the expression of CREB, p-CREB, p-GSK3, and Bcl-2 was slightly decreased. In addition, all the p-GSK3/GSK3, p-CREB/CREB, p-NF-κB/NF-κB, and p-BAD/BAD ratios were significantly reduced in favipiravir-treated cells. These results implied that the antiviral effectivity of favipiravir against PPRV is mediated by the JAK/STAT and PI3K/AKT pathways and that favipiravir has potential for use as an effective antiviral agent against PPRV.

Quantitative Analysis of Lysine Acetylation in Vero Cells Infected With Peste Des Petits Ruminants Virus

2021 ◽  
Author(s):  
Xuelian Meng ◽  
Xueliang Zhu ◽  
Rui Zhang ◽  
Zhidong Zhang
Keyword(s):  
Rna Virus ◽  
Small Ruminants ◽  
Vero Cells ◽  
Host Cells ◽  
Lysine Acetylation ◽  
Peste Des Petits Ruminants ◽  
Bioinformatics Analyses ◽  
Protein Protein Interaction ◽  
Quantification Analysis ◽  
And Function

Abstract Background: Peste des petits ruminants virus (PPRV) is a negative-stranded RNA virus belonging to the Paramyxoviridae family and causes acute, highly contagious disease in small ruminants. Lysine acetylation plays central role in regulating gene expression. However, the extent and function of lysine acetylation in host cells during PPRV infection remains unknown. Methods: Lysine acetylation of PPRV-infected Vero cells was tested and differentially expressed lysine acetylation was found. The acetylated peptides were enriched using specific antibody and labeled with demethylation. Proteins with acetylation sites were identified. Subsequently, intensive bioinformatics analysis of succinylome of PPRV-infected Vero cells was were performed. In this study, intensive proteomic quantification analysis of the proteome and acetylome of PPRV-infected Vero cells was performed using dimethylation labeling-based quantitative proteomics. Results: We identified 4729 cellular proteins and 1068 proteins with 2641 modification sites quantifiable detected by mass spectrometry, of which 304 proteins with 410 acetylation sites were significantly acetylated in response to PPRV infection. Bioinformatics analyses revealed that the differentially acetylated proteins mainly participated in carbohydrate catabolic and DNA metabolic process, and were associated with multifarious functions, suggesting that intracellular activities were extensively changed after PPRV infection. Protein-protein interaction (PPI) network of the identified proteins further indicated that a variety of chaperone and ribosome processes were modulated by acetylation. Conclusions: To our knowledge, this is the first study on acetylome in host cell infected with PPRV. It provides an important baseline to future study the roles of acetylation in the host response to PPRV replication.

scholarly journals Peste des petits ruminants virus infection of goats in Bangladesh: Pathological investigation, molecular detection and isolation of the virus

1970 ◽  
Vol 28 (1) ◽  
pp. 1-7 ◽  
Author(s):  
MA Rahman ◽  
I Shadmin ◽  
M Noor ◽  
R Parvin ◽  
EH Chowdhury ◽  
...  
Keyword(s):  
Lymph Node ◽  
Vero Cells ◽  
Viral Rna ◽  
Sub Saharan Africa ◽  
Tissue Homogenate ◽  
Clinical Samples ◽  
Peste Des Petits Ruminants ◽  
Rt Pcr ◽  
Sub Saharan ◽  
Bronchial Lymph Node

Peste des petits ruminants (PPR), an economically important morbillivirus infection of sheep and goats, is widely distributed in sub-Saharan Africa, Middle East and western and southern Asia including Bangladesh.  A small flock of Black Bengal goats contracted PPR following introduction of new animals. A pathological investigation was conducted on the outbreak, the viral RNA corresponding to F gene was detected by RT-PCR and the virus was isolated in Vero cells. Out of 37 goats 19 (51%) developed clinnical disease, of which 5 (13.5%) died. Goats under one year of age had highest morbidity and mortality with typical signs and lesions of PPR. Viral RNA should be detected in mesenteric and bronchial lymph node tissues. Typical cytopathic effects (CPE) in Vero cells following inoculation of lymph node tissue homogenate were visible at the third passage. However, the replication of virus in cel culture was detected by RT-PCR at the first and second passage in the absence of visible CPE. RT-PCR appears to be a very useful and sensitive tool not only for the detection of PPR virus in clinical samples but also for monitoring the growth or virus in cell culture following inoculation.DOI: http://dx.doi.org/10.3329/bvet.v28i1.8808 Bangl. vet. 2011. Vol. 28, No. 1, 1–7

Antibacterial and Cytotoxic Effects of Silver Nanoparticles on Staphylococcus aureus and Normal Vero Cells

2019 ◽  
Vol 22 (10) ◽  
pp. 184-190
Author(s):  
Rasha Hadi Saleh ◽  
Entisar J. Al-Mukhtar ◽  
Zaytoon A. Al-Khafaji ◽  
Mohammed H. Al Hasnawy ◽  
Huda H. Al-Hasnawy
Keyword(s):  
Staphylococcus Aureus ◽  
Silver Nanoparticles ◽  
Vero Cells ◽  
Cytotoxic Effects

Phytochemical screening, cytotoxicity and anti-inflammatory activities of the leaf extracts from Lawsonia inermis of Indian origin to explore its potential for medicinal uses

2020 ◽  
Vol 16 ◽  
Author(s):  
Anju Manuja ◽  
Nitu Rathore ◽  
Shalki Chaudhary ◽  
Balvinder Kumar
Keyword(s):  
Nitric Oxide ◽  
Cytotoxic Activity ◽  
Cytotoxic Effect ◽  
Vero Cells ◽  
Nitric Oxide Production ◽  
Lawsonia Inermis ◽  
Leaf Extracts ◽  
Oxide Production ◽  
Indian Origin ◽  
Anti Inflammatory

Background: Lawsonia inermis Linn popularly known as the Henna has played an important role in ayurvedic or natural herbal medicines. The presence of phyto-constituents in henna, that may affect the animal or human health adversely, need to be elucidated for L. inermis Linn species grown in India. Introduction: Introduction: The aim of this research was to perform phytochemical, cytotoxicity and anti-inflammatory studies to understand the potential of leaves of Lawsonia inermis of Indian origin to provide a way forward for therapeutic use in medicine. Methods: We assessed the phytochemical profile for presence of phyto-constituents (alkaloids, carbohydrates, glycosides, steroids, flavonoids, saponins, tannins, proteins/amino acids and gums/mucilage) from various extracts of the plant leaves’. The extracts were further purified by column chromatography for the isolation of plant constituents and monitored by TLC, analyzed by Fourier transform infrared FT-IR spectroscopy, H1NMR, and GC-MS analysis. Fractions were assessed for cytotoxicity and anti-inflammatory properties at various concentrations. We assessed the anti-inflammatory activity by nitric oxide production in various leaf extracts determined by Griess assay. Results: All the spectral results suggest that the compounds from the extract contain aromatic nucleus and OH group along with methoxy group, allyl as well as vinyl group. Fractions of chloroform/methanolic (7:3) leaf extract of Lawsonia inermis confirmed the presence of the two constituents i.e. fraxetin and 1(3H)-isobenzofuranone. We observed significant difference in cytotoxicity at higher concentrations in methanol and chloroform:methanol (8:2) leaf extracts (p>0.05), we could not find any significant differences amongst other leaf extracts at different concentrations. Some leaf extracts have potential cytotoxic activity on vero cells. Reducing the chloroform concentration during extraction decreases the cytotoxic effect on the cells. The nitric oxide levels decreased from 1000 µg/ml concentration to lower concentrations with varying degree. Overall the highest nitric oxide production by CHCl3 (70%)/ MeOH (30%) was observed amongst various fractions at different concentrations. Conclusion: The phytochemical, cytotoxicity and anti-inflammatory studies indicating the potential of leaves of the plant to provide a way further for their use in medicine. Fraxetin 1(3H)-isobenzofuranone structures were confirmed in fractions of CHCl3 (70%)/ MeOH (30%) extract as observed as a potent constituents. Some leaf extracts have potential cytotoxic activity on vero cells. Reducing the chloroform concentration during extraction decreases the cytotoxic effect on the cells.The cytotoxicity studies indicates the presence of cytotoxic compounds in some of these extracts, warranting research for fabrication of suitable formulations comprising these constituents to reduce its dose/toxicity for the use of beneficial effects of the plant components.

scholarly journals Bufadienolides from the Skin Secretions of the Neotropical Toad Rhinella alata (Anura: Bufonidae): Antiprotozoal Activity against Trypanosoma cruzi

Molecules ◽  
2021 ◽  
Vol 26 (14) ◽  
pp. 4217
Author(s):  
Candelario Rodriguez ◽  
Roberto Ibáñez ◽  
Luis Mojica ◽  
Michelle Ng ◽  
Carmenza Spadafora ◽  
...  
Keyword(s):  
Vero Cells ◽  
Chemical Diversity ◽  
Hydroxyl Group ◽  
Antitrypanosomal Activity ◽  
The Family ◽  
Toad Venom ◽  
Skin Secretions ◽  
Pharmacological Potential ◽  
First Time ◽  
Mcf 7

Toads in the family Bufonidae contain bufadienolides in their venom, which are characterized by their chemical diversity and high pharmacological potential. American trypanosomiasis is a neglected disease that affects an estimated 8 million people in tropical and subtropical countries. In this research, we investigated the chemical composition and antitrypanosomal activity of toad venom from Rhinella alata collected in Panama. Structural determination using mass spectrometry (MS) and nuclear magnetic resonance (NMR) spectroscopy led to the identification of 10 bufadienolides. Compounds identified include the following: 16β-hydroxy-desacetyl-bufotalin-3-adipoyl-arginine ester (1), bufotalin (2), 16β-hydroxy-desacetyl-bufotalin-3-pimeloyl-arginine ester (3), bufotalin-3-pimeloyl-arginine ester (4), 16β-hydroxy-desacetyl-bufotalin-3-suberoyl-arginine ester (5), bufotalin-3-suberoyl-arginine ester (6), cinobufagin-3-adipoyl-arginine ester (7), cinobufagin-3-pimeloyl-arginine ester (8), cinobufagin-3-suberoyl-arginine ester (9), and cinobufagin (10). Among these, three new natural products, 1, 3, and 5, are described, and compounds 1–10 are reported for the first time in R. alata. The antitrypanosomal activity assessed in this study revealed that the presence of an arginyl-diacid attached to C-3, and a hydroxyl group at C-14 in the structure of bufadienolides that is important for their biological activity. Bufadienolides showed cytotoxic activity against epithelial kidney Vero cells; however, bufagins (2 and 10) displayed low mammalian cytotoxicity. Compounds 2 and 10 showed activity against the cancer cell lines MCF-7, NCI-H460, and SF-268.

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