Sequential Adaptive Mutations Enhance Efficient Vector Switching by Chikungunya Virus and Its Epidemic Emergence

The adaptation of Chikungunya virus (CHIKV) to a new vector, the Aedes albopictus mosquito, is a major factor contributing to its ongoing re-emergence in a series of large-scale epidemics of arthritic disease in many parts of the world since 2004. Although the initial step of CHIKV adaptation to A. albopictus was determined to involve an A226V amino acid substitution in the E1 envelope glycoprotein that first arose in 2005, little attention has been paid to subsequent CHIKV evolution after this adaptive mutation was convergently selected in several geographic locations. To determine whether selection of second-step adaptive mutations in CHIKV or other arthropod-borne viruses occurs in nature, we tested the effect of an additional envelope glycoprotein amino acid change identified in Kerala, India in 2009. This substitution, E2-L210Q, caused a significant increase in the ability of CHIKV to develop a disseminated infection in A. albopictus, but had no effect on CHIKV fitness in the alternative mosquito vector, A. aegypti, or in vertebrate cell lines. Using infectious viruses or virus-like replicon particles expressing the E2-210Q and E2-210L residues, we determined that E2-L210Q acts primarily at the level of infection of A. albopictus midgut epithelial cells. In addition, we observed that the initial adaptive substitution, E1-A226V, had a significantly stronger effect on CHIKV fitness in A. albopictus than E2-L210Q, thus explaining the observed time differences required for selective sweeps of these mutations in nature. These results indicate that the continuous CHIKV circulation in an A. albopictus-human cycle since 2005 has resulted in the selection of an additional, second-step mutation that may facilitate even more efficient virus circulation and persistence in endemic areas, further increasing the risk of more severe and expanded CHIK epidemics

Yeast expressed recombinant Hemagglutinin protein of Novel H1N1 elicits neutralising antibodies in rabbits and mice

Currently available vaccines for the pandemic Influenza A (H1N1) 2009 produced in chicken eggs have serious impediments viz limited availability, risk of allergic reactions and the possible selection of sub-populations differing from the naturally occurring virus, whereas the cell culture derived vaccines are time consuming and may not meet the demands of rapid global vaccination required to combat the present/future pandemic. Hemagglutinin (HA) based subunit vaccine for H1N1 requires the HA protein in glycosylated form, which is impossible with the commonly used bacterial expression platform. Additionally, bacterial derived protein requires extensive purification and refolding steps for vaccine applications. For these reasons an alternative heterologous system for rapid, easy and economical production of Hemagglutinin protein in its glycosylated form is required. The HA gene of novel H1N1 A/California/04/2009 was engineered for expression in Pichia pastoris as a soluble secreted protein. The full length HA- synthetic gene having α-secretory tag was integrated into P. pastoris genome through homologous recombination. The resultant Pichia clones having multiple copy integrants of the transgene expressed full length HA protein in the culture supernatant. The Recombinant yeast derived H1N1 HA protein elicited neutralising antibodies both in mice and rabbits. The sera from immunised animals also exhibited Hemagglutination Inhibition (HI) activity. Considering the safety, reliability and also economic potential of Pichia expression platform, our preliminary data indicates the feasibility of using this system as an alternative for large-scale production of recombinant influenza HA protein in the face of influenza pandemic threat

Inflammatory Cytokine Expression Is Associated with Chikungunya Virus Resolution and Symptom Severity

The Chikungunya virus infection zones have now quickly spread from Africa to parts of Asia, North America and Europe. Originally thought to trigger a disease of only mild symptoms, recently Chikungunya virus caused large-scale fatalities and widespread economic loss that was linked to recent virus genetic mutation and evolution. Due to the paucity of information on Chikungunya immunological progression, we investigated the serum levels of 13 cytokines/chemokines during the acute phase of Chikungunya disease and 6- and 12-month post-infection follow-up from patients of the Italian outbreak. We found that CXCL9/MIG, CCL2/MCP-1, IL-6 and CXCL10/IP-10 were significantly raised in the acute phase compared to follow-up samples. Furthermore, IL-1β, TNF-α, Il-12, IL-10, IFN-γ and IL-5 had low initial acute phase levels that significantly increased at later time points. Analysis of symptom severity showed association with CXCL9/MIG, CXCL10/IP-10 and IgG levels. These data give insight into Chikungunya disease establishment and subsequent convalescence, which is imperative to the treatment and containment of this quickly evolving and frequently re-emerging disease

Progressive hemorrhage and myotoxicity induced by echis carinatus venom in murine model: neutralization by inhibitor cocktail of n,n,n `,n `-tetrakis (2-pyridylmethyl) ethane-1,2-diamine and silymarin

Viperbite is often associated with severe local toxicity, including progressive hemorrhage and myotoxicity, persistent even after the administration of anti-snake venom (ASV). In the recent past, investigations have revealed the orchestrated actions of Zn2+ metalloproteases (Zn(2+)MPs), phospholipase A(2)s (PLA(2)s) and hyaluronidases (HYs) in the onset and progression of local toxicity from the bitten site. As a consequence, venom researchers and medical practitioners are in deliberate quest of potent molecules alongside ASV to tackle the brutal local manifestations induced by aforesaid venom toxins. Based on these facts, we have demonstrated the protective efficacy of inhibitor cocktail containing equal ratios of N,N,N', N'-tetrakis (2-pyridylmethyl) ethane-1,2-diamine (TPEN) and silymarin (SLN) against progressive local toxicity induced by Echis carinatus venom (ECV). In our previous study we have shown the inhibitory potentials of TPEN towards Zn(2+)MPs of ECV (IC50: 6.7 mu M). In this study we have evaluated in vitro inhibitory potentials of SLN towards PLA(2)s (IC50: 12.5 mu M) and HYs (IC50: 8 mu M) of ECV in addition to docking studies. Further, we have demonstrated the protection of ECV induced local toxicity with 10 mM inhibitor cocktail following 15, 30 min (for hemorrhage and myotoxicity); 60 min (for hemorrhage alone) of ECV injection in murine model. The histological examination of skin and thigh muscle sections taken out from the site of ECV injection substantiated the overall protection offered by inhibitor cocktail. In conclusion, the protective efficacy of inhibitor cocktail is of high interest and can be administered locally alongside ASV to treat severe local toxicity

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Not AvailableTwo types of male sterile lines isolated in marigold were evaluated for their effective contribution for heterosis exploitation of biochemical components. Among seven male sterile lines used as seed parents in the study, three are petaloid sterile lines and four are apetaloid sterile lines. Three fertile pure lines, were used as pollen parents evaluated in different cross combination with all the 7 sterile lines. Line versus tester mean squares for combining ability were significant for all the biochemical components. The results of combining ability revealed that, total carotenoids, zeaxanthin and lutein appeared to be governed by non-additive gene action. Estimates of general combining ability effects showed that parent IIHRMO 9-7 was a good general combiner for lutein and total carotenoid. For total carotenoids and lutein, the highest positive values of heterotic effects were recorded in hybrid combination IIHRMO 9-8× IIHRMO 12-12 relative to the better parentNot Availabl

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Not AvailableThirty three genotypes of African marigold (Tagetes erecta) were evaluated for 27 characters contributing for yield, carotenoid and lutein. Based on the method of propagation, the study material comprised of three vegetative propagated and thirty seed propagated genotypes. Phenotypic coefficient of variability (PCV) and genotypic coefficient of variation (GCV) was maximum for fresh petal meal per flower and dry petal meal per flower. High broad sense heritability coupled with high genetic gain was observed for number of secondary branches, fresh petal meal per flower, dry petal meal per flower, dry weight per flower, flower weight per plant, flower number per plant, zea-xanthin content, lutein content, total carotenoid content, seed number per flower, 100 seed weight, number of seeds per gram and shelf life. It indicates that most likely the heritability due to additive gene effects and selection will be effective. Efficiency of hybrid seed production depends on ability of seed production. Attempts have been made to study seed production in both apetaloid and petaloid sterile lines. Variability analysis was attempted to analyse the characters contributing for seed yield and the results suggested that the characters that contribute for flower yield also contributed for seed yield.Not Availabl

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Not AvailableThirty three genotypes of African marigold (Tagetes erecta) were evaluated for 27 characters contributing for yield, carotenoid and lutein. Based on the method of propagation, the study material comprised of three vegetative propagated and thirty seed propagated genotypes. Phenotypic coefficient of variability (PCV) and genotypic coefficient of variation (GCV) was maximum for fresh petal meal per flower and dry petal meal per flower. High broad sense heritability coupled with high genetic gain was observed for number of secondary branches, fresh petal meal per flower, dry petal meal per flower, dry weight per flower, flower weight per plant, flower number per plant, zea-xanthin content, lutein content, total carotenoid content, seed number per flower, 100 seed weight, number of seeds per gram and shelf life. It indicates that most likely the heritability due to additive gene effects and selection will be effective. Efficiency of hybrid seed production depends on ability of seed production. Attempts have been made to study seed production in both apetaloid and petaloid sterile lines. Variability analysis was attempted to analyse the characters contributing for seed yield and the results suggested that the characters that contribute for flower yield also contributed for seed yield.Not Availabl

Thermal effects on rapid microwave synthesized Co: ZnAl2O4 spinel nanoparticles

A rapid (10 min) microwave-assisted synthesis of CoxZn(1-x)Al2O4 nanoparticles using metal organic precursors is reported for the first time. The nanoparticles formed at 185 degrees C were phase-pure in the asprepared form with an average crystallite size of similar to 5 nm. Sample with x = 0.0025 showed intense photoluminescence (PL) emission and was used to study the effect of annealing temperature on the structural and optical properties. Annealing in air until 1200 degrees C gradually (i) increased the crystallite size to as much as 31 nm (ii) change of colour tone from colourless to blue which has been attributed to the diffusion of Co2+ in host lattice (iii) increase in intensity of Co2+ bands in the diffuse reflectance spectra (DRS). The emission spectra comprised of the broad T-4(1)(P-4)/(4)A(2)(F-4) spin allowed transition pertaining to tetrahedral Co2+ at similar to 649 nm whose intensity increased with annealing temperature. The crystal field splitting parameter (D-q) and the Racah parameter B, were calculated from the excitation spectra and the Tanabe-Sugano diagrams. The novelty of this work lies in the simplicity and swiftness of the microwaveassisted synthesis that can be extended to other ceramic phosphors suitable for lasing and sensing applications. (C) 2017 Elsevier B.V. All rights reserved

Rapid annealing: A novel processing technique for Cr:ZnAl2O4 nanoparticles

Nanocrystalline Cr: ZnAl2O4 was synthesized for the first time by rapid thermal annealing the corresponding hydroxide precipitate in just 10 min. The XRD patterns showed that synthesis between 500 degrees C and 700 degrees C had smaller crystallites (similar to 2.4 - 4 nm), while at 800 degrees C and 900 degrees C, had similar to 10 nm crystallites. HRTEM images of the 900 degrees C sample showed the well-developed lattice fringes confirming the high crystallinity. The particles were polyhedral having an average size of similar to 16 nm. The diffuse reflectance spectra showed characteristic (4)A(2g) -> T-4(1g) and (4)A(2g) -> T-4(2g) transitions, while the photoluminescence emission spectrum comprised of the zero phonon line (R-line) along with other multi-phonon side bands that are characteristic of the spin - parity forbidden transitions of the Cr3+ ion. The emission spectra and photoluminescence lifetimes are comparable with reports on sensing and imaging applications. Our work demonstrates the suitability of this technique to quickly synthesise phosphors for a variety of applications. (C) 2017 Elsevier B. V. All rights reserved