Measles virus considers an important cause of child morbidity and mortality in some areas as Africa. Ribavirin's activity as a nucleoside analog can disclose the surprisingly broad spectrum action against several RNA viruses under laboratory cell culture conditions. The Current study aimed to investigate the antiviral activity of ribavirin Nano gold particles (AuNPs) against measles virus on vero cell line. Ribavirin- AuNPs was prepared, characterization and the cytotoxicity of ribavirin, AuNPs and ribavirin -AuNPs were tested on vero cells using MTT assay. Antiviral activiry of ribavirin, AuNPs and ribavirin- AuNPswere determined on vero cells using simultaneous, pre-infection and post-infection protocols. Results indicated safety of ribavirin and ribavirin-AuNPs on vero cells, there was a reduction by 78.1% when vero cells treated with ribavirin -AuNPs at 99.5µg/ml while, the viral reduction was 25.4% when ribavirin 500 µg /ml was used for the same viral concentration. Our results concluded that ribavirin - AuNPs had a higher antiviral activity with lower dose than ribavirin alone and the maximal activity showed when it used after the virus infection.
Background Rift Valley Fever Virus (RVFV) is an arbovirus, a zoonotic disease that resurfaces as a potential hazard beyond geographic boundaries. Fever that can proceed to encephalitis, retinitis, hemorrhagic fever, and death is the main manifestation observed in human infections. RVFV has no authorized medication. The RNA interference (RNAi) gene silencing pathway is extremely well conserved. By targeting specific genes, small interfering RNA (siRNA) can be used to suppress viral replication. The aim of this study was to design specific siRNAs against RVFV and evaluate their prophylactic and antiviral effects on the Vero cells. Methods and results Various siRNAs were designed using different bioinformatics tools. Three unique candidates were tested against an Egyptian sheep cell culture-adapted strain BSL-2 that suppressed RVFV N mRNA expression. SiRNAs were transfected a day before RVFV infection (pre-transfection), and 1 h after the viral infection (post-transfection), and were evaluated to detect the silencing activity and gene expression decrease using real-time PCR and a TCID50 endpoint test. The degree of N protein expression was determined by western blot 48 h after viral infection. D2 which targets the (488–506 nucleotides), the middle region of RVFV N mRNA was the most effective siRNA at 30 nM concentration, it almost eliminates N mRNA expression when utilized as antiviral or preventive therapy. siRNAs had a stronger antiviral silencing impact when they were post-transfected into Vero cells. Conclusion Pre and post-transfection of siRNAs significantly reduced RVFV titer in cell lines, offering novel and potentially effective anti-RVFV epidemics and epizootics therapy.
Legumes are very important species including herbs, shrubs and trees, they are cultivated worldwide for different economic uses.Parkiais genus belongs to the Legumefamily, Fabaceae. The most common known species are P. biglobosaBenth (African locust bean) and P. roxburghii (Tree Bean)which were introduced to Egypt since 1890. The population of the tree is rapidly declining as result conservation efforts is needed to prevent it from extinction. In an attempt to conserve this genetic resource, tissue culture studies were carried toestablish a callus culture of both plants from different explants and regeneration of the developed callus and to study the effect of explant type, media composition on the callus production and the regeneration. Apical bud,axillary bud and root explants from in vitro germinated seedlings of P. biglobosaBenthwere cultured on MS and B5 medium supplemented with (0.4., 0.8, 1.2, 1.6 or 2.0 mg l-1 NAA) + Glutamine 0.5 mg l-1 + Proline 0.5 mg l-1 + Casein 0.3 mg l-1 + 2,4D 2.5 mg l-1. Each medium was supplemented with 30 g l-1 sucrose, 3 g l-1 agar and PH 5.8.for callus induction .and (0.4.,0.8,1.2,1.6 or 2.0 mg l-1 NAA) + Glutamine 0.5 mg l-1 + Proline 0.5 mg l-1 + Casein 0.3 mg l-1 + 6-BAP 2 mg l-1. Each medium was supplemented with 30 g l-1 sucrose, 3 g l-1 agar and PH 5.8.for regeneration.The study presented here demonstrates the successful attempt at regeneration of plantlets of P. biglobosaBenth via indirect organogenesis.It can be concluded that the medium composition has a significant effect on the calli production together with the type of the explant which showed relatively higher significance. Seeds germination stage apical bud explants media 8 followed by media 10 showed the most favorable mean performances values for the studied characters, Number of days to callus initiation (day) , Callus weight (mg), Dead calli percentage (%),Albino plants percentage (%),Green plants percentage (%) and Root percentage (%). Moreover, axillary bud, showed remarkable increase in the mean performance of average. However, the overall response of MS medium was found to be superior to that of B5 medium. Explants cultured on MS medium fortified with combinations of 2,4-D and BAP induced rapidly proliferating calli that turned more friable and nodular.
Rift Valley Fever Virus (RVFV) is an arbovirus, zoonotic pathogen that repeatedly reinvigorates with a possible threat across geographic borders. There are no authorized drugs for RVFV. RNA interference gene silencing pathway is a highly conserved mechanism. Specific and different small interfering RNAs (siRNA) have been designed against RVFV Nucleoprotein genes by different bioinformatics tools to investigate their antiviral potentialities in the Vero cell line. Real-time PCR besides Endpoint assay was used to determine the silencing activity of siRNA and reduction in gene expression as all designed siRNAs abrogate mRNA replication by more than 90% by RT-PCR and significant inhibition in RVFV replication in cell line when used as antiviral as well prophylactic therapeutics. Western blot analysis was utilized for protein abundance detection after 48 hours. Consequently, specific siRNA can inhibit RVFV replication in a cell-based assay, introducing novel and promising RVFV epidemics and epizootics therapeutics.
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