A single amino acid change resulting in loss of fluorescence of eGFP in a viral fusion protein confers fitness and growth advantage to the recombinant vesicular stomatitis virus

Using a recombinant vesicular stomatitis virus encoding eGFP fused in-frame with an essential viral replication protein, the phosphoprotein P, we show that during passage in culture, the virus mutates the nucleotide C289 within eGFP of the fusion protein PeGFP to A or T, resulting in R97S/C amino acid substitution and loss of fluorescence. The resultant non-fluorescent virus exhibits increased fitness and growth advantage over its fluorescent counterpart. The growth advantage of the non-fluorescent virus appears to be due to increased transcription and replication activities of the PeGFP protein carrying the R97S/C substitution. Further, our results show that the R97S/C mutation occurs prior to accumulation of mutations that can result in loss of expression of the gene inserted at the G-L gene junction. These results suggest that fitness gain is more important for the recombinant virus than elimination of expression of the heterologous gene

A single amino acid change resulting in loss of fluorescence of eGFP in a viral fusion protein confers fitness and growth advantage to the recombinant vesicular stomatitis virus

Using a recombinant vesicular stomatitis virus encoding eGFP fused in-frame with an essential viral replication protein, the phosphoprotein P, we show that during passage in culture, the virus mutates the nucleotide C289 within eGFP of the fusion protein PeGFP to A or T, resulting in R97S/C amino acid substitution and loss of fluorescence. The resultant non-fluorescent virus exhibits increased fitness and growth advantage over its fluorescent counterpart. The growth advantage of the non-fluorescent virus appears to be due to increased transcription and replication activities of the PeGFP protein carrying the R97S/C substitution. Further, our results show that the R97S/C mutation occurs prior to accumulation of mutations that can result in loss of expression of the gene inserted at the G-L gene junction. These results suggest that fitness gain is more important for the recombinant virus than elimination of expression of the heterologous gene

Interferon Inducible Protein IFI35 Negatively Regulates RIG-I Antiviral Signaling and Supports Vesicular Stomatitis Virus Replication

In a genome-wide siRNA screen, we recently identified the interferon (IFN) inducible protein 35 (IFI35, also known as IFP35) as a factor required for VSV infection. Studies reported here were conducted to further understand the role and requirement of IFI35 in VSV infection. Consistent with the siRNA screening data, we found that depletion of IFI35 led to reduced VSV replication at the level of viral gene expression. Although no direct interaction of IFI35 with the viral replication machinery was observed, we found that IFI35 negatively regulated the host innate immune response and rescued poly(I:C)-induced inhibition of VSV replication. Promoter-driven reporter gene assays demonstrated that IFI35 overexpression suppressed the activation of IFNβ and ISG56 promoters, whereas its depletion had opposite effect. Further investigation revealed that IFI35 specifically interacted with RIG-I and negatively regulated its activation through mechanisms that include: (i) suppression of dephosphorylation (activation) of RIG-I and (ii) proteasome-mediated degradation of RIG-I via K48-linked ubiquitination. Overall, the results presented here suggest a novel role for IFI35 in negative regulation of RIG-I mediated antiviral signaling, which will have implications for diseases associated with excessive immune signaling

Growth, Nutrient Utilization and Carcass Characteristics in Broiler Chickens Fed Raw and Alkali Processed Solvent Extracted Karanj (Pongamia glabra) Cake as Partial Protein Supplement

An experiment was conducted to evaluate the effect of dietary incorporation of raw and alkali [1.5% NaOH and 3% Ca (OH)2, w/w] treated solvent extracted karanj (Pongamia glabra) cake (SKC) on growth performance and carcass characteristics in broiler chickens during 0 to 6 weeks of age. A basal reference diet was formulated containing soybean meal (SBM) as the major protein source. Another six isonitrogenous and isocaloric test diets were formulated incorporating SKC, 1.5% NaOH treated SKC (NaOH-SKC) and 3% Ca (OH)2 treated SKC (Ca (OH)2-SKC) at 6.43 or 5.5% during starter phase and 12.86 or 11.0% during finisher phase replacing SBM nitrogen of reference diet at 12.5 and 25%, respectively. Each diet was offered ad. libitum to 4 replicates of 10 chicks each. The body weight gain and feed efficiency of broilers fed 6.43% NaOH-SKC incorporated diet was comparable with reference diet during 0 to 28d. However, weight gain and feed efficiency reduced in all karanj cakes incorporated diet during 29 to 42d. The overall weight gain during 0 to 42d was significantly higher in the reference diet. Significantly higher nutrient intake was observed in the reference group except 6.43% NaOH-SKC diet during both the balance trials and 6.43% Ca (OH)2-SKC during 2nd trial. The percent retention of DM, N, Ca, P and GE did not differ significantly. Higher liver weight was observed due to dietary incorporation of SKC and NaOH-SKC at 25%, and Ca (OH)2-SKC. The gizzard weight was significantly higher in SKC incorporated diet, at both the levels of replacement compared to reference diet. The breast yield lowered significantly when soybean meal nitrogen was replaced with processed or unprocessed karanj cake at 25% level. The findings suggested that 1.5% NaOH (w/w) treated SKC could be incorporated upto 6.43% level, replacing 12.5% of soybean nitrogen of reference diet in broiler chicken upto 4 weeks of age

Induction of Interferon and Interferon Signaling Pathways by Replication of Defective Interfering Particle RNA in Cells Constitutively Expressing Vesicular Stomatitis Virus Replication Proteins▿

We show here that replication of defective interfering (DI) particle RNA in HEK293 cells stably expressing vesicular stomatitis virus (VSV) replication proteins potently activates interferon (IFN) and IFN signaling pathways through upregulation of IFN-β promoter, IFN-stimulated response element (ISRE) promoter, and NF-κB promoter activities. Replication of DI particle RNA, not mere expression of the viral replication proteins, was found to be critical for induction of IFN and IFN signaling. The stable cells supporting replication of DI RNA described in this report will be useful in further examining the innate immune signaling pathways and the host cell functions in viral genome replication

In-situ deposited CdS NPs on pH induced fully exfoliated layered titanate-biopolymeric composite and its photocatalytic activity

Herein, pH mediated in-situ formation, stabilization, exfoliation of layered titanate has been explored for various types of exfoliation of the stacked layered titanate (H2Ti2O5·H2O). Based on exfoliation, a CdS NPs/exfoliated layered titanate based crosslinked biopolymeric nanohybrid has been developed via in-situ approach. The synthetic strategy of polymeric hybrid-composite follows simultaneous increase of the degree of polymerisation, in-situ controlled decomposition/hydrolysis of the pre-stage stacked layered titanate and its full exfoliation, followed by the growth of CdS NPs on the surface of exfoliated titanate nanosheets at 75 °C. Further, the investigation on photocatalytic degradation has been carried out in details to correlate the advantages of exfoliation and deposition of CdS NPs on stacked/unstacked titanate towards degradation of toxic 2,4-dichlorophenol under sunlight irradiation

Photocatalytic TiO2 incorporated PVDF-co-HFP UV-cleaning mixed matrix membranes for effective removal of dyes from synthetic wastewater system via membrane distillation

View PDF Journal of Environmental Chemical Engineering Volume 9, Issue 5, October 2021, 105904 Photocatalytic TiO2 incorporated PVDF-co-HFP UV-cleaning mixed matrix membranes for effective removal of dyes from synthetic wastewater system via membrane distillation Author links open overlay panelAnshulYadavacPreranaSharmaacAsit BaranPandabcVinod KumarShahiac a Membrane Science and Separation Technology Division, CSIR-Central Salt & Marine Chemicals Research Institute, Gijubhai Badheka Marg, Bhavnagar 364002, India b Advanced Materials and Processes Division, CSIR-National Metallurgical Laboratory, Jamshedpur 831007, India c Academy of Scientific and Innovative Research (AcSIR), Ghaziabad 201002, India Received 9 March 2021, Revised 15 June 2021, Accepted 17 June 2021, Available online 24 June 2021. Editor: Despo Kassinos crossmark-logo https://doi.org/10.1016/j.jece.2021.105904 Get rights and content Highlights • UV-cleaning PVDF-co-HFP/TiO2 mixed matrix membrane for DCMD. • Controllable large-scale synthesis of porous TiO2 sheets by spray drying process. • Effective removal of MB and CR dye from wastewater system by membrane distillation. • High performance DCMD for five days without significant flux and rejection decline. Abstract We report membranes with UV-cleaning properties by incorporating photocatalytic porous TiO2 sheets (PTS) (0–5 wt%) in the PVDF-co-HFP matrix. PTS were synthesized using controllable large-scale synthesis protocols by spray drying followed by calcination. These membranes were assessed for the removal of Congo red (CR) and methylene blue (MB) from synthetic textile industry wastewater system (CR: 100 mg l−1 + MB: 100 mg l−1 + 4% NaCl) via direct contact membrane distillation (DCMD) configuration and their UV-cleaning properties. The PTS were characterized by FE-SEM, TEM and different spectral techniques, while membranes were assessed by surface morphology (FE-SEM, AFM), thermal stability, hydrophobicity, permeate flux, and dye rejection performance. The surface morphology study of PTS revealed its micro-sized sheet with the porous surface. With increased PTS concentration in the membrane matrix, the flux and dye rejection of mixed matrix membranes improved. Suitably optimized PT3 (3 wt% PTS in PVDF-co-HFP matrix) membrane showed ~100% dye removal efficiency (MB and CR) and 6.1 kg m−2 h−1 vapour flux. Long run (5 days) DCMD experiments showed >91% flux recovery ratio (FRR) after UV-cleaning. This study suggests the suitability of the prepared membranes to treat textile wastewater and their UV-cleaning properties

Biarsenical Labeling of Vesicular Stomatitis Virus Encoding Tetracysteine-Tagged M Protein Allows Dynamic Imaging of M Protein and Virus Uncoating in Infected Cells▿

A recombinant vesicular stomatitis virus (VSV-PeGFP-M-MmRFP) encoding enhanced green fluorescent protein fused in frame with P (PeGFP) in place of P and a fusion matrix protein (monomeric red fluorescent protein fused in frame at the carboxy terminus of M [MmRFP]) at the G-L gene junction, in addition to wild-type (wt) M protein in its normal location, was recovered, but the MmRFP was not incorporated into the virions. Subsequently, we generated recombinant viruses (VSV-PeGFP-ΔM-Mtc and VSV-ΔM-Mtc) encoding M protein with a carboxy-terminal tetracysteine tag (Mtc) in place of the M protein. These recombinant viruses incorporated Mtc at levels similar to M in wt VSV, demonstrating recovery of infectious rhabdoviruses encoding and incorporating a tagged M protein. Virions released from cells infected with VSV-PeGFP-ΔM-Mtc and labeled with the biarsenical red dye (ReAsH) were dually fluorescent, fluorescing green due to incorporation of PeGFP in the nucleocapsids and red due to incorporation of ReAsH-labeled Mtc in the viral envelope. Transport and subsequent association of M protein with the plasma membrane were shown to be independent of microtubules. Sequential labeling of VSV-ΔM-Mtc-infected cells with the biarsenical dyes ReAsH and FlAsH (green) revealed that newly synthesized M protein reaches the plasma membrane in less than 30 min and continues to accumulate there for up to 2 1/2 hours. Using dually fluorescent VSV, we determined that following adsorption at the plasma membrane, the time taken by one-half of the virus particles to enter cells and to uncoat their nucleocapsids in the cytoplasm is approximately 28 min

Membrane distillation crystallization for simultaneous recovery of water and salt from tannery industry wastewater using TiO2 modified poly (vinylidene fluoride-co-hexafluoropropylene) nanocomposite membranes

Membrane distillation crystallization (MDCr) is a promising technology for simultaneous water recovery and salt crystallization from tannery industry wastewater. We synthesized TiO2 modified poly(vinylidene fluoride-cohexafluoropropylene) (PVDF-HFP) nanocomposite flat sheet membranes to treat tannery industry wastewater by MDCr. The influence of TiO2 nanoparticles morphologies on membrane performance (salt rejection and water flux) was investigated. The nanocomposite membranes were analyzed for surface morphology, thermal stability, porosity, water flux, and salt rejection. It was demonstrated that the TiO2 modified PVDF-HFP nanocomposite membrane showed improved hydrophobicity, porosity and number of pores; thus, water flux significantly increased without any deterioration in salt rejection. Among different membranes, decanoic acid-modified TiO2 nanoparticles (bipyramidal morphology) nanocomposite membrane (M-1B) showed 6.1 kg m(-2) h(-1) water flux with similar to 100% salt rejection with mixed synthetic salt solution (1% NaCl and Na2SO4, each) feed. Using real tannery industry wastewater, the M-1B membrane showed 5.9 kg m(-2) h(-1) water flux and 99.97% salt rejection. The crystallization of salts revealed cubic NaCl crystals and monoclinic Na2SO4 crystals in the wastewater. The XRD spectra revealed a high purity of salts and ruled out any co-crystallization. These findings suggested the potential candidature of the M-1B membrane for the treatment of tannery industry wastewater by the MDCr process. The recovered salt and water can also be re-used in the tannery industry

Structural tailoring of ceria nanoparticles for fabricating fouling resistant nanocomposite membranes with high flux distillation

This study stems from the morphological design of ceria (CeO2) filler into the membrane matrix, which might provide a varied method for building nanocomposite membranes with superior performance. We synthesized nanostructured CeO2 with controlled size and shape using aqueous ceric ammonium carbonate complex (precursor) and decanoic acid under reflux and hydrothermal conditions. Poly(vinylidene fluoride-co-hexafluoropropylene) (PVDF-co-HFP)/CeO2 nanocomposite membranes were fabricated through the phase inversion technique for desalination via direct contact membrane distillation (DCMD) using the synthesized CeO2 of various morphologies. The influence of CeO2 morphology on the performance of the membrane was investigated by various morphological and spectral techniques along with transmembrane flux and salt rejection. Among the fabricated membranes, (18% PVDF-co-HFP/1%) CeO2 (synthesized through hydrothermal route with 1:16 ratio of ceric ammonium nitrate to decanoic acid) -M-HT1:16 membrane was adjudged the most suitable with 5.33 L m(-2) h(-1) permeate flux and 99.98% salt rejection for desalination of 8% NaCl solution at 60 degrees C feed temperature. The smooth spherical morphology of CeO2 nanoparticles improved surface roughness, membrane porosity, and hydrophobicity for the M-HT1:16 membrane. A mathematical model was also proposed which describe the vapor flux profile of the DCMD process. The model was validated by experimental data with reasonable accuracy. Exceptional thermal stability, anti-fouling nature, along with feasible scale-up make MHT1:16 nanocomposite membrane a promising candidate for seawater desalination by the DCMD process