Background: Replication of influenza virus to high titer is a prerequisite for successful Cell-based vaccine production. Entry of virus into the Cell depends on the cleavage of the hemagglutinin precursor (HA0) protein mediated by trypsin. Objectives: The aim of the present study was to apply a technique to establish MDCK/FX manipulated Cell, which may provide a new platform for developing influenza vaccine based on the Cell culture approaches. Methods: Chicken embryo FX expressed into the pCDNA3. 1 vector was transfected into the MDCK Cell line. The longevity of the generated Cell and the viable Cell density were evaluated for 17 passages prior to virus inoculation. Then, the ability of MDCK/FX Cell for efficient replication of H9N2 influenza virus was evaluated by viral titration and quantitative RT-PCR. Results: RT-PCR data revealed that FX was stably expressed in the Cell after the subsequent passages without any change in the rate of culture’ s confluency. Growth kinetic of H9N2 virus analysis demonstrated that MDCK/FX Cell supported high-titer growth of the virus in which the viral titer is comparable to the virus grown in MDCK Cells supplemented with TPCK-trypsin. Quantification of influenza infectious particles in the Cell culture revealed the equivalents viral RNA copies and viral titers. Conclusions: The results indicated a potential application for the MDCK/FX in influenza virus replication procedure and related studies.

Studies of interaction between viruses and mitochondria have shown that they can affect the mitochondria and induce mitochondrial alterations. Sometimes this interaction leads to induction or inhibition of apoptosis in infected Cells. Interaction between adenoviruses and mitochondria showed the first evidence that a viral infection could affect the Cell and induces apoptotic process. In addition, linkage between human diseases and mitochondrial dysfunction has been revealed. A Previously work showed that poliovirus infection led to decrease in total Cellular respiration induced by inhibition of mitochondrial electron transport chain. We have screened different human viruses to observe their effect on mitochondrial respiration using Oxygen Electrode (OE). Here we report the effect of influenza A virus (IAV) on mitochondrial Cell respiration. The addition of antimycin A (AA) to respiring Cells completely blocked Cellular respiration that indicates nearly all of the Cellular consumption of oxygen is attributed to the mitochondria. Our result showed human influenza A virus (IAV) decreases total Cell respiration in infected MDCK Cells.  

Objective(s): The protective effect of ethyl acetate fraction (EAF) of Boerhavia diffusa roots against Tacrolimus (TAC) induced nephrotoxicity was studied using MDCK Cell lines. Materials and Methods: Ethanolic root extract of B. diffusa was fractionated using the liquid-liquid partition method. The cytotoxic effect of TAC and protective effect of EAF co-treatment were studied in MDCK Cell lines by measuring ROS, LPO, and NO levels; collagen accumulation, effect on mitochondrial membrane integrity and Cell cycle analysis were studied. The active component in EAF was quantified by HPLC analysis. Results: TAC induced toxicity, leading to apoptosis and necrosis, was significantly reduced (P<0. 001) in EAF co-treatment, with reversal of Cell cycle arrest and reduced Cell population at sub G0/G1 phase. Further, ROS (P<0. 05), LPO and NO (P<0. 001), were significantly reduced with EAF co-treatment compared with TAC individually treated Cells. TAC induced mitochondrial membrane integrity loss was found to be significantly reduced in co-treated Cells, as measured by rhodamine123 (P<0. 05) and translocation of cytochrome c (P<0. 001) from nucleus to cytoplasm, and caspase 3 release (P<0. 001). The same was confirmed through annexin-FITC and PI staining (P<0. 05) with reduced apoptotic and necrotic death in co-treated population. Interestingly, EAF co-treatment decreased collagen accumulation (P<0. 001) with significant increase in the Cell survival of tubular epithelial Cells. HPLC analysis showed the presence of Quercetin (87. 5 mg/g) in EAF, which may be responsible for the nephroprotective role. Conclusion: Thus, these results provide sound evidence that EAF may be an effective adjuvant therapy to prevent nephrotoxicity induced by TAC.

Background: Tannic acid plays a significant role in modifying silver nanoparticles due to its unique properties and functionalities. Objective(s) : In this work, demonstrate the tannic acid modified silver nanoparticles exhibit different cytotoxicity compared to other silver nanoparticles due to the presence of the tannic acid coating. The tannic acid modification with silver nanoparticles can decrease their ability to inhibit the proliferation of MDCK Cells line. Tannic acid itself has antioxidant properties and can scavenge free radicals, which may help mitigate the cytotoxic effects of silver nanoparticles. Materials and Methods: A green biosynthesis was used to prepare successful product and establish a tannic acid and silver nanoparticles (TA-AgNPs). TA-AgNPs was prepared in several concentrations (3.1, 6.25, 12.5, 25, 50) μg/mL and evaluated for cytotoxicity effect in vitro normal MDCK Cell line. on the surface of silver nanoparticles tannic acid was effectively modified and characterized via UV Visible Spectroscopy, Fourier transform infrared (FTIR) spectroscopy, AFM, XRD, SEM and TEM. Results: The observations with SEM indicated that produced TA-AgNPs had a spherical shape with a mean particle size of 13.55±5.9 nm. The cytotoxic effect and The anti-proliferative activity of TA-AgNPs treated MDCK Cells showed the TA-AgNPs have less cytotoxicity and high bioavailability rate (90.66±0.7212) than AgNPs (90.65±0.8864)in the same concentration (50 μg/mL). Conclusion: This study revealed that tannic acid modified silver nanoparticles have a highly effective metal nanoparticles with promising new applications. Because of its significant unique properties, tannic acid may be referred to as the cross-linker in the future.

Introduction: Inactivated influenza vaccines are traditionally produced in chicken embryonated eggs but its limitations in producing the required doses in pandemic outbreaks quickly enough has made searching for alternative modes of production necessary. The use of Cell culture-based vaccine production is one way of overcoming the limitations of the egg-based method and securing a more rapid response. Although Vero Cells are suitable for production of influenza vaccine, but their anchorage-dependency limits their production capability. In this study adherent Vero and MDCK Cells were transfected with human siat7e gene in order to convert anchorage-dependent Cells to those capable of growing in suspension. Methods: Human siat7e gene was amplified with primers containing restriction sites for Xho I and Hind III and the product was cloned into pEGFP-N1 vector upstream of GFP sequence. The Cells were transfected with the construct containing the siat7e gene and a medium containing G418 was used to select stably transfected Cells which were then evaluated using inverted immunofluorescence microscopy. Results: Anchorage-dependent Cells exhibited changes in Cell-Cell adhesion and Cell spreading behavior following transfection. Vero Cells showed a higher longevity compared to MDCK Cells as viability of the latter Cells declined after 50 h. Conclusion: The data showed that adherent Vero Cells can successfully be converted to anchorage-independent Cells capable of growing in suspension through transfection with human siat7e gene.

The influenza viruses are major etiologic agents of human respiratory infections, and inflict a sizable health and economic burden. This study examines the antiinfluenza virus activity of hydroalcoholic extract of olive leaves (OLHE). Olive leaves were collected from gardens around the city of Shiraz, characterized, dried, ground to powder, and its hydroalcoholic extract was prepared. The influenza viruses were isolated from patients and characterized by standard antiinfluenza sera. Virucidal effects of OLHE (10-1 to 103 μg/ml) were examined in pretreatment, treatment and incubation protocols using quantal assay after incubation for 72 h. All experiments were performed three times in quadruplicates. Pretreatment of the Cell line with OLHE for one hour followed by the addition of the virus was associated with virucidal effects (1 to 1000 μg/ml). OLHE added one hour after incubation of the virus with Cell did not show antiviral effects. OLHE incubated with the virus for one hour, and then added to the Cell line did have antiviral activity (1 to 1000 μg/ml). The findings indicate that antiviral activity of OLHE occurred extraCellularly, probably by changing the properties of membrane of the virus, rather than that of the Cell, to prevent the virus from attaching and penetrating the Cell line.