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        The effective role of positive charge saturation in bioluminescence color and thermostability of firefly luciferase

        Alipour, Bagher Said,Hosseinkhani, Saman,Ardestani, Sussan K.,Moradi, Ali Korean Society of Photoscience 2009 Photochemical & photobiological sciences Vol.8 No.6

        Luciferases are the enzymes that catalyze the reactions that produce light in bioluminescence. The bioluminescence color of firefly luciferases is determined by the luciferase structure and assay conditions. Amongst different beetle luciferases, those from phrixothrix rail-road worm with a unique additional residue (Arg353) emit red bioluminescence color naturally. Insertion of $Arg^{356}$ in Lampyris turkestanicus luciferase changed the emitted light to red with a bimodal bioluminescence spectrum. By insertion and substitution of positively-charged residues, different specific mutation (E354R/$Arg^{356}$, E354K/$Arg^{356}$, E354R, E354K) lead to changes of the bioluminescence color. Bioluminescence emission spectra indicate that substitution of E354 by R along with insertion of $Arg^{356}$ produces a luciferase that emits red light with a single peak bioluminescence spectrum. The comparison of mutants with native luciferase shows that mutations of firefly luciferase resulted in structural and functional thermostability. Comparative study of native and mutant luciferase (E354R/$Arg^{356}$) by intrinsic and extrinsic fluorescence, CD spectropolarimetry, and homology modeling revealed mutation brought about an increase in content of secondary structure and globular compactness of L. turkestanicus luciferase. On the other hand, $pK_a$ of amino acids in the flexible loop decreased upon introducing of positive charges.

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        TB Trifusion Antigen Adsorbed on Calcium Phosphate Nanoparticles Stimulates Strong Cellular Immunity in Mice

        Yousef Amini,Bagher Moradi,Mohsen Tafaghodi,Zahra Meshkat,Kiarash Ghazvini,Mahdi Fasihi-Ramandi 한국생물공학회 2016 Biotechnology and Bioprocess Engineering Vol.21 No.5

        Among various vaccine candidates, subunit vaccines play an important role in immune protection against tuberculosis (TB). Calcium phosphate (CP) is considered as a strong inorganic adjuvant due to its great potential in increasing immune responses. The purpose of this study was to evaluate specific immune responses following the administration of trifusion-CP nanoparticles. The physiochemical properties of these nanoparticles, including morphology, particle size, zeta potential and adsorption rate, were measured in vitro. Subcutaneous immunization was performed three times on days 0, 14, and 28. Two weeks after the last administration, IFN-gamma, IL-4, and TGF-beta levels were measured by indirect enzyme linked immunosorbent assay (ELISA). The trifusion protein was successfully adsorbed onto calcium phosphate nanoparticles. The mean sizes of the resultant trifusion- CPN and CPN were 97.84 ± 12.08 and 67 ± 11.85 nm, respectively. CPN containing trifusion had stronger ability to induce IFN-gamma than the control groups. IL-4 and TGF-beta secretions in trifusion and trifusion-CPN groups were higher than those in the PBS group. However, there was no significant (p > 0.05) difference in IL-4 and TGFbeta concentrations between trifusion group and trifusion- CPN group. Therefore, calcium phosphate nanoparticles are good candidates for immunization against TB because antigen can be easily adsorbed onto CPN and strong cellular immune responses against CPN-antigen can be stimulated.

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        Application of Taguchi method to satellite image fusion for object-oriented mapping of Barchan dunes

        Vahid Moosavi,Seyed Rashid Fallah Shamsi,Hamidreza Moradi,Bagher Shirmohammadi 한국지질과학협의회 2014 Geosciences Journal Vol.18 No.1

        In this study, a wavelet-based method and some standard fusion methods were performed on IRS Panchromatic and multi-spectral images to benefit from its spatial and spectral advantages simultaneously. Selection of the best fusion parameters using a trial and error approach is often very time-consuming. So the best wavelet-based fusion parameters were determined using optimization Taguchi method and the importance degree of these parameters in wavelet-based image fusion efficiency were figured out. As a preliminary result, image fusion with Db4 as the wavelet type in which images were decomposed in level 4 with the Maximum and Mean functions as fusion methods for approximations and details is the best wavelet-based fusion method. The method has been compared with several standard fusion techniques according to spectral and spatial characteristics of participating bands. The results indicate that the wavelet based method presents a better spectral performance than the standard ones (R = 0.96), but PCA fusion method has a better spatial performance (RMSE = 1.06). This study also indicates that Taguchi method can be used accurately in order to optimize the performance of wavelet-based image fusion and determine the importance degree of parameters used in this method. Finally, a pixel-based and an object-oriented classification procedure have been employed to delineate barchan dunes using the fused images. The results show that object-oriented approach outperforms pixel-based approach in classifying barchan dunes (Kappa = 0.73 for pixel-based and Kappa = 0.87 for object-oriented). It is also demonstrated that the wavelet-based fused images are better inputs for classification processes than the others.

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