Three-dimensional (3D) models for the 65-kDa activated Cry4A and Cry4B $\delta$-endotoxins from Bacillus thuringiensis subsp. israelensis that are specifically toxic to mosquito-larvae were constructed by homology modeling, based on atomic coordinates of the Cry1Aa and Cry3Aa crystal structures. They were structurally similar to the known structures, both derived 3D models displayed a three-domain organization: the N-terminal domain (I) is a seven-helix bundle, while the middle and C-terminal domains are primarily comprise of anti-parallel $\beta$-sheets. Circular dichroism spectroscopy confirmed the secondary structural contents of the two homology-based Cry4 structures. A structural analysis of both Cry4 models revealed the following: (a) Residues Arg-235 and Arg-203 are located in the interhelical 5/6 loop within the domain I of Cry4A and Cry4B, respectively. Both are solvent exposed. This suggests that they are susceptible to tryptic cleavage. (b) The unique disulphide bond, together with a proline-rich region within the long loop connecting ${\alpha}4$ and ${\alpha}5$ of Cry4A, were identified. This implies their functional significance for membrane insertion. (c) Significant structural differences between both models were found within domain II that may reflect their different activity spectra. Structural insights from this molecular modeling study would therefore increase our understanding of the mechanic aspects of these two closely related mosquito-larvicidal proteins.
Crystal structural analyses of the API-TLCK complex revealed that the ${\epsilon}$-amino group (NZ) of the lysyl part of TLCK forms hydrogen bonds with OD1 of $Asp^225$ which is a substrate specificity determinant of API, OG of $Ser^214$, O of $Ser^214$, OG1 of $Thr^189$, and O of $Thr^189$ l89/. The ${\beta}$-carboxyl oxygen of $Asp^225$ forms hydrogen bonds with the NE1 of $Trp^182$. From these observations, it is thought that besides $Asp^225$, $Thr^189$, $Ser^214$, and $Trp^182$ may also contribute to the steric specificity for lysine and high proteolytic activity of API. The side-chain hydroxyl groups of $Thr^189$ and $Ser^214$ were removed to elucidate the role of these hydrogen bonds in the $S_1$-pocket. The $k_{cat}$/$K_m$ of T189V, S214A, and T189V.S214A were decreased to 1/4, 1/3, and 1/46, respectively, of the value for native API. The decreased activities were mainly due to the increase of $K_m$. The CD and fluoroscence spectra of the three mutants were similar to those of wild-type API. With regards to the kinetic parameters ($K_i\;and\;k_2$) of mutants for the reaction involving TLCK and DFP, $k_2$decreased by increase of $K_1$ only. These results suggest that the decreased catalytic activity of these mutants is caused by the partial loss of the hydrogen bond network in the $S_1$-pocket. On the other hand, the similarity of enzymatic properties between W182F and the native enzyme suggests that the hydrogen bond between OD2 of $Asp^225$ and NE1 of $Trp^182$ is not directly related to the reaction of $Asp^225$ with the substrate.
The Transactions of the Korean Institute of Electrical Engineers C
/
v.55
no.9
/
pp.434-441
/
2006
The effect of pressureless-sintered temperature on the densification behavior, mechanical and electrical properties of the $SiC-ZrB_2$ electroconductive ceramic composites was investigated. The $SiC-ZrB_2$ electroconductive ceramic composites were pressureless-sintered for 2 hours at temperatures in the range of $1,750{\sim}1,900[^{\circ}C]$, with an addition of 12[wt%] of $Al_2O_3+Y_2O_3$(6:4 mixture of $Al_2O_3\;and\;Y_2O_3$) as a sintering aid. The relative density and mechanical properties are increased markedly at temperatures in the range of $1,850{\sim}1,900[{^\circ}C]$. The relative density, flexural strength, vicker's hardness and fracture toughness showed the highest value of 81.1[%], 230[MPa], 9.88[GPa] and $6.05[MPa\;m^{1/2}]$ for $SiC-ZrB_2$ composites of $1,900[{^\circ}C]$ sintering temperature at room temperature respectively. The electrical resistivity was measured by the Pauw method in the temperature ranges from $25[{^\circ}C]\;to\;700[{^\circ}C]$, The electrical resistivity showed the value of $1.36{\times}10^{-4},\;3.83{\times}10^{-4},\;3.51{\times}10^{-4}\;and\; 3.2{\times}10^{-4}[{\Omega}{\cdot}cm]$ for SZ1750, SZ1800, SZ1850 and SZ1900 respectively at room temperature. The electrical resistivity of the composites was all PTCR(Positive Temperature Coefficient Resistivity). The resistance temperature coefficient showed the value of $4.194{\times}10^{-3},\;3,740{\times}10^{-3},\;2,993{\times}10^{-3},\;3,472{\times}10^{-3}/[^{\circ}C}$ for SZ1750, SZ1800, SZ1850 and SZ1900 respectively in the temperature ranges from $25[{\circ}C]\;to\;700[{\circ}C]$, It is assumed that because polycrystallines such as recrystallized $SiC-ZrB_2$ electroconductive ceramic composites, contain of porosity and In Situ $YAG(Al_5Y_3O_{12})$ crystal grain boundaries, their electrical conduction mechanism are complicated. In addition, because the condition of such grain boundaries due to $Al_2O_3+Y_2O_3$ additives widely varies with sintering temperature, electrical resistivity of the $SiC-ZrB_2$ electroconductive ceramic composites with sintering temperature also varies with sintering condition. It is convinced that ${\beta}-SiC$ based electroconductive ceramic composites for heaters or ignitors can be manufactured by pressureless sintering.
Mohamed, Shereen H;Mahmoud, Nora F;Mohamed, Aly F;Kotb, Nahla S
Asian Pacific Journal of Cancer Prevention
/
v.16
no.14
/
pp.5635-5641
/
2015
Background: Cirrhosis is regarded as a possible end stage of many liver diseases, including viral infection. It occurs when healthy liver tissue becomes damaged and is replaced by scar tissue and finally may lead to hepatocellular carcinoma. Interferons (IFNs)are two general categories, type I and II. Type I includes one beta interferon and over 20 different alpha interferons. Alpha interferons are very similar in how they work, interacting with other proteins on cells like receptors. The main objective of this study was to compare Mx gene productivity post different cell line treatment with imported and Egyptian biosimilar locally produced IFNs, as well as the efficacy of those tested IFNs. Also, an assessment was made of sensitivity of different cell lines as alternatives to that recommended for evaluation of antiviral activity. Materials and Methods: Different cell lines (Vero, MDBK and Wish) were employed to evaluate cytotoxicity using the MTT assay. Antiviral activity was evaluated compared with standard IFN against VSV, Indiana strain -156, on tested rh-IFNs (imported; innovated and Egyptian biosimilar locally produced IFNs) in the pre-treated cell lines previously mentioned. The virus was propagated in the Wish cell line as recommended. Finally we estimated up-regulation of the Mx gene as a biomarker. Results: Data recorded revealed that test IFNs were safe in test cell lines. Viability was around 100%. Locally tested interferon did not realize the international potency limits, while the imported one was accepted compared with the standard IFN. These results were the same either using infectivity titer reduction assay or crystal violet staining of residual non- infected cells. Mx protein production was cell type related and confirmed by the detected Mx gene expressed in imported and locally produced IFN pre-treated cell lines. The expression of the gene was arranged in the order of Vero> wish > MDBK for the imported IFN, while for the Egyptian biosimillar locally produced one it was MDBK> Vero> wish. With regard to the antiviral activity there was a significant difference of imported IFN potency compared with the locally produced IFN (P<0.05), the IFN potential (antiviral activity) was not cell line related and showed non-significant difference for each separate product. Conclusions: Vero cells can be used as an alternative cell line for evaluation of IFN potency in case of unavailable USP recommended cell lines. Alternative potency evaluation assay could be used and proved significant difference in IFN potency in case of local and imported agents. Evaluation of antiviral activity could be used in parallel to viral infectivity reduction assay for better accuracy. Mx gene can be used as a marker for IFN potential.
Alginates are found in marine brown seaweeds and in extracellular biofilms secreted by some bacteria. Previously, we reported an oligoalginate lyase from Sphingomonas sp. MJ-3 (MJ3-Oal) that had an exolytic activity and protein sequence homology with endolytic polymannuronate (polyM) lyase in the N-terminal region. In this study, the MJ3-Oal was tested for both exolytic and endolytic activity by homology modeling using the crystal structure of Alg17c from Saccharophagus degradans 2-40T. The tyrosine residue at the $426^{th}$ position, which possibly formed a hydrogen bond with the substrate, was mutated to phenylalanine. The FPLC profiles showed that MJ3-Oal degraded alginate quickly to monomers as a final product through the oligmers, whereas the Tyr426Phe mutant showed only exolytic alginate lyase activity. $^1H$-NMR spectra also showed that MJ3-Oal degraded the endoglycosidic bond of polyM and polyMG (polymannuronate-guluronate) blocks. These results indicate that oligoalginate lyase from Sphingomonas sp. MJ-3 probably catalyzes the degradation of both exo- and endo-glycosidic bonds of alginate.
The Transactions of the Korean Institute of Electrical Engineers C
/
v.55
no.10
/
pp.467-474
/
2006
The effect of pressureless-sintered temperature on the densification behavior, mechanical and electrical properties of the $SiC-TiB_2$ electroconductive ceramic composites was investigated. The $SiC-TiB_2$ electroconductive ceramic composites were pressureless-sintered for 2 hours at temperatures in the range of $1,750{\sim}1,900[^{\circ}C]$, with an addition of 12[wt%] $Al_2O_3+Y_2O_3(6:4\;mixture\;of\;Al_2O_3\;and\;Y_2O_3)$ as a sintering aid. The relative density, flexural strength, vicker's hardness and fracture toughness showed the highest value of 84.92[%], 140[MPa], 4.07[GPa] and $3.13[MPa{\cdot}m^{1/2}]$ for $SiC-TiB_2$ composites of $1,900[^{\circ}C]$ sintering temperature at room temperature respectively. The electrical resistivity was measured by the Pauw method in the temperature ranges from $25[^{\circ}C]\;to\;700[^{\circ}C]$. The electrical resistivity showed the value of $5.51{\times}10^{-4},\;2.11{\times}10^{-3},\;7.91{\times}10^{-4}\;and\;6.91{\times}10^{-4}[\Omega{\cdot}cm]$ for ST1750, ST1800, ST1850 and ST1900 respectively at room temperature. The electrical resistivity of the composites was all PTCR(Positive Temperature Coefficient Resistivity). The resistance temperature coefficient showed the value of $3.116{\times}10^{-3},\;2.717{\times}10^{-3},\;2.939{\times}10^{-3},\;3.342{\times}10^{-3}/[^{\circ}C]$ for ST1750, ST1800, ST1850 and ST1900 respectively in the temperature ranges from $25[^{\circ}C]\;to\;700[^{\circ}C]$. It is assumed that because polycrystallines, such as recrystallized $SiC-TiB_2$ electroconductive ceramic composites, contain of porosity and In Situ $YAG(Al_5Y_3O_{12})$ crystal grain boundaries, their electrical conduction mechanism are complicated. In addition, because the condition of such grain boundaries due to $Al_2O_3+Y_2O_3$ additives widely varies with sintering temperature, electrical resistivity of the $SiC-TiB_2$ electroconductive ceramic composites with sintering temperature also varies with sintering condition. It is convinced that ${\beta}-SiC$ based electroconductive ceramic composites for heaters or ignitors can be manufactured by pressureless sintering.
The plain coarse pottery from the Unjeonri Bronze Age relic sites in the Cheonan, Korea were studied on the basis of clay mineralogy, geochemistry and archaegeological interpretations. For the research, the potteries are utilized at the analysis for 6 pieces of plain coarse potteries. Color of the these potteries are mainly light brown, partly shows the yellowish brown to reddish brown. The interior, surface and inside of the pottery appear as different colors in any cases. Original source materials making the Unjeonri potteries are used of mainly sandy clay soil with extreme coarse grained irregularly quartz and feldspar. The magnetic susceptibility of the Unjeonri pottery range from 0.20 to 1.20. And the Unjeonri soil's magnetic susceptibility agree almost with 0.20 to 1.30. In the same magnetization of soil and pottery, the results revealed that the Unjeonri soil and low material of pottery are same produced by identical source materials. The Unjeonri potteries and soil are very similar patterns with all characteristics of soil mineralogy, geochemical evolution trend. The result seems to be same relationships between the behavior and enrichment patterns on the basis of a compatible and a incompatible elements. Consequently, the Unjeonri potteries suggest that made the soil to be distributed in the circumstance of the relic sites as the raw material are high in a greater part. In the Unjeonri soil, the kaolinite is common occurred minerals. However, in the Unjeonri pottery, the kaolinite was not detected in all broken pieces. The kaolinite was presumed to destroy crystal structure during the firing processes of over $550^{\circ}C$. The quartz is phase transition from ${\alpha}$-quartz to ${\beta}$-quartz at $573^{\circ}C$, but the Unjeonri pottery did not investigated any phase transition evidences of quartz. The chorite was detected within the mostly potteries and soils. As the results, the Unjeonri potteries can be interpreted by not experiencing a firing temperature over $800^{\circ}C$. The colloidal and cementing materials between the quartz and low materials during the heating did not exist in the internal part of the potteries. An any secondary compounds by heating does not appear within the crack to happen during the dry of the pottery. The hyphae group are kept as it is with the root tissue of an organic matters to live in the swampy land. In the syntheses of all results, the general firing condition to bake and make the Unjeonri pottery is presumed from $550^{\circ}C$ to $800^{\circ}C$. However, the firing condition making the Unjeonri pottery can be different firing temperature partially in one pottery. Even, the some part of the pottery does not take a direct influence on the fire.
The $LiMn_2O_4$ cathodes for lithium ion battery were synthesized from various precursors of manganese oxides and manganese hydroxides. As the first step, nanosized precursors such as ${\alpha}-MnO_2$ (nano-sticks), ${\beta}-MnO_2$ (nano-rods), $Mn_3O_4$ (nano-octahedra), amorphous $MnO_2$(nano-spheres), and $Mn(OH)_2$ (nano-plates) were prepared by a hydrothermal or a precipitation method. Spinel $LiMn_2O_4$ with various sizes and shapes were finally synthesized by a solid-state reaction method from the manganese precursors and LiOH. Nano-sized (500 nm) octahedron $LiMn_2O_4$ showed high capacities of 107 mAh $g^{-1}$ and 99 mAh $g^{-1}$ at 1 C- and 50 C-rate, respectively. Three dimensional octahedral crystallites exhibit superior electrochemical characteristics to the other one-dimensional and two-dimensional shaped $LiMn_2O_4$ nanoparticles. After 500 consecutive charge discharge battery cycles at 10 C-rate with the nano-octahedron $LiMn_2O_4$ cathode, the capacity retention of 95% was observed, which is far better than any other morphologies studied in this work.
Journal of the korean academy of Pediatric Dentistry
/
v.23
no.3
/
pp.640-658
/
1996
The purpose of this study was to examine the mechanism of improving acid resistance of Nd-YAG laser irradiated tooth enamel and determine the most effective energy density for improving acid resistance. The bovine tooth enamel were lased with a pulsed Nd-YAG laser. The energy densities of exposed laser beam were varied from 10 to $70\;J/cm^2$. To investigate the degree of improving acid resistance by irradiation, all the samples were submerged to demineralize in 0.5 N $HClO_4$ solution for 1 minute. After 1 minute, 0.05 % $LaCl_3$ was added to the solution for interrupting the demineralization reaction. The amounts of dissolved calcium and phosphate in the solution were measured by using an atomic absorption spectrophotometer and the UV/VIS spectrophotometer, respectively. To examine the mechanism of improving acid resistance, X-ray diffraction analysis, infrared spectroscopy, and scanning electron microscopy were taken. The X-ray diffraction pattern of the samples were obtained in the $10^{\circ}{\sim}80^{\circ}2{\theta}$ range with $Cu-K{\alpha}$ radiation using M18HF(Mac Science Co.) with X-ray diffractometer operating at 40 KV and 300 mA. The infra-red spectra of the ground samples in 300 mg KBr pellets 10 mm diameter were obtained in the $4000cm^{-1}\;to\;400cm^{-1}$ range using JASCO 300E spectrophotometer. The scanning electron microscopy was carried out using JSM6400(JEOL Co.) with $500{\sim}2000$ times magnification. The results were as follow 1. The concentration of calcium dissolved from laser irradiated enamel with $50J/cm^2$ was significantly lesser than that of unlased control group (p<0.05) 2. From the result of the X-ray diffraction analysis, $\beta$-TCP, which increases acid solubility, was identified in lased enamel but the diffraction peaks of (002) and (004) became sharp with increasing energy density of laser irradiation. This means that the crystals in lased samples were grown through the c-axis and subsequently, the acid solubility of enamel decreased. 3. The a-axis parameter was slightly increased by laser irradiation, whereas the c-axis parameter was almost constant except for a little decrease at $50J/cm^2$. 4. In the infra-red spectra of lased enamels, phosphate bands ($600{\sim}500cm^{-1}$), B-carbonate bands (870, $1415{\sim}1455cm^{-1}$), and A-carbonate band ($1545cm^{-1}$) were observed. The amounts of phosphate bands and the B-carbonate bands were reduced, on the other hand, the amount of the A-carbonate band was increased by increase the energy density. 5. The SEM experiments reveal that the surface melting and recrystallization were appeared at $30J/cm^2$ and the cracks were observed at $70J/cm^2$. From above results, It may be suggested that the most effective energy density for improving acid resistance of tooth enamel with the irradiation of Nd-YAG laser was $50J/cm^2$. The mechanism of improving acid resistance were reduction of permeability due to surface melting and recrystallization of lased enamel and reduction of acid solubility of enamel due to decrease of carbonate content and growth of crystal.
Kim, Kyong-Mok;Lee, Byung-Wook;Lee, Dong-Wook;Kim, Jeong-Su;Jang, Yeong-Do;Bang, Chan-Seok;Baek, Jong-Hun;Lee, In-Su
The Korean Journal of Nuclear Medicine Technology
/
v.14
no.2
/
pp.33-37
/
2010
Purpose: In the early stage of using PET/CT, it was used to damper revision but recently shows that CT with MDCT is commonly used and works well for an anatomical diagnosis. This hospital makes the accuracy and convenience more higher in the diagnosis and evaluate of coronary heart disease through concurrently running myocardial perfusion SPECT examination, myocardial PET examination with FDG, and CT coronary artery CT angiography(coronary CTA) used PET/CT with 64-slice. This report shows protocol and image based on results from about 400 coronary heart disease examinations since having 64 channels PET/CT in July 2007. Materials and Methods: An Equipment for this examination is 64-slice CT and Discovery VCT (DVCT) that is consisted of PET with BGO ($Bi_4Ge_3O_{12}$) scintillation crystal by GE health care. First myocardial perfusion SPECT with pharmacologic stress test to reduce waiting time of a patient and get a quick diagnosis and evaluation, and right after it, myocardial FDG PET examination and coronary CTA run without a break. One-stop evaluation protocol of ischemic heart disease is as follows. 1)Myocardial perfusion SPECT with pharmacologic stress: A patient is injected with $^{99m}Tc$-MIBI 10 mCi and does not have any fatty food for myocardial PET examination and drink natural water with ursodeoxcholic acid 100 mg and we get SPECT image in an hour. 2)Myocardial FDG PET: To reduce blood fatty content and to increase uptake of FDG, we used creative oral glucose load using insulin and Acipimox to according to blood acid content. A patient is injected with $^{18}F$-FDG 5 mCi for reduction of his radiation exposure and we get a gated image an hour later and get delay image when we need. 3) Coronary CTA: The most important point is to control heart rate and to get cooperation of patient's breath. In order to reduce a heart rate of him or her below 65 beats, let him or her take beta blocker 50 mg ~ 200 mg after a consultation with a doctor about it and have breath-practices then have the examination. Right before the examination, we spray isosorbide dinitrate 3 to 5 times to lower tension of bessel wall and to extension a blood wall of a patient. It makes to get better the shape of an anatomy. At filming, a patient is injected CT contrast with high pressure and have enough practices before the examination in order to have no problem. For reduction of his radiation exposure, we have to do ECG-triggered X-ray tube modulation exposure. Results: We evaluate coronary artery stenosis through coronary CTA and study correlation (culprit vessel check) of a decline between stenosis and perfusion from the myocardial perfusion SPECT with pharmacologic stress, coronary CTA, and can check viability of infarction or hibernating myocardium by FDG PET. Conclusion: The examination makes us to set up a direction of remedy (drug treatment, PCI, CABG) because we can estimate of effect from remedy, lesion site and severity. In addition, we have an advantage that it takes just 3 hours and one-stop in that all of process of examinations run in succession and at the same time. Therefore it shows that the method is useful in one stop evaluation of ischemic heart disease.
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