Journal of the Korea Academia-Industrial cooperation Society
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v.18
no.1
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pp.295-301
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2017
Botulinum neurotoxin (BoNT/A) is a neurotoxin that selectively attacks the peripheral cholinergic nerve endings. It is produced by Gram -positive, endospore-forming strict anaerobic bacteria, Clostridium botulinum. Since BoNT/A could be a biothreat agent, as well as a contaminator of food and water supplies, the development of sensitive assays for toxin detection and potent antitoxin for the treatment of intoxication is necessary. In this study, for the purpose of producing monoclonal antibodies (mAbs) that are capable of neutralizing Botulinum neurotoxin type A (BoNT/A), scFv (single-chain variable domain fragment) libraries from the rabbit antisera against BoNT/A was fused to a human IgG. The resulting recombinant scFvIgG antibody protein was expressed in stable cell lines and was purified using a protein A affinity chromatography. The efficacy of scFvIgG mAb was confirmed by ELISA and was evaluated for the neutralization of BoNT/A in vivo. Such an in vivo toxin neutralization assay was performed using mice. Although scFvIgG antibody proteins (10 ug) failed to fully protect the mice challenged with BoNT/A (100,000 $LD_{50}$), it significantly prolonged the survival time. These results suggest that scFvIgG mAb may be capable of neutralizing BoNT/A single-chain variable domain fragment.
The cylindrical ${\gamma}$-alumina pellets were prepared by forming, hydration, drying and calcination after mixing amorphous alumina and pore generating agent with water. Concentration of Fe(NO$_3$)$_3$ㆍ9$H_2O$ that was catalyst precursor was fixed and made mixing solution that changed concentration of $CH_3$COOH in range of 2.5~20%, and here ${\gamma}$-alumina pellets were immerged and were hydrothermaly treated for 3 h at $200^{\circ}C$. And then we investigated creation and change of crystal, pore characteristics, $N_2$ adsorption and desorption isotherms, changes of acid site and mechanical strengths etc. According to the concentration of $CH_3$COOH, the crystals grew to acicular shape of 0.5~2${\mu}m$ length, and crystal structure showed the pseudo-boehmite structure. When hydrothermaly treated in 10% $CH_3$COOH solution, pore volume between 100~1000 $\AA$ was highest by 0.86 cc/g, and width of hysteresis curved line due to $N_2$ adsorption/desorption appeared as was smallest. When concentration of $CH_3$COOH was in range of 5~15%, new C-H functional groups were formed. Mechanical strength of pellets was highest by 1.35 MPa when $CH_3$COOH concentration was 2.5%.
Glioblastoma multiforme (GBM) is one of the most common brain malignancies and has a very poor prognosis. Recent evidence suggests that the presence of cancer stem cells (CSC) in GBM and the rare CSC subpopulation that is resistant to chemotherapy may be responsible for the treatment failure and unfavorable prognosis of GBM. A garlic-derived compound, Z-ajoene, has shown a range of biological activities, including anti-proliferative effects on several cancers. Here, we demonstrated for the first time that Z-ajoene specifically inhibits the growth of the GBM CSC population. CSC sphere-forming inhibition was achieved at a concentration that did not exhibit a cytotoxic effect in regular cell culture conditions. The specificity of this inhibitory effect on the CSC population was confirmed by detecting CSC cell surface marker CD133 expression and biochemical marker ALDH activity. In addition, stem cell-related mRNA profiling and real-time PCR revealed the differential expression of CSC-specific genes, including Notch, Wnt, and Hedgehog, upon treatment with Z-ajoene. A proteomic approach, i.e., reverse-phase protein array (RPPA) and Western blot analysis, showed decreased SMAD4, p-AKT, 14.3.3 and FOXO3A expression. The protein interaction map (http://string-db.org/) of the identified molecules suggested that the AKT, ERK/p38 and $TGF{\beta}$ signaling pathways are key mediators of Z-ajoene's action, which affects the transcriptional network that includes FOXO3A. These biological and bioinformatic analyses collectively demonstrate that Z-ajoene is a potential candidate for the treatment of GBM by specifically targeting GBM CSCs. We also show how this systemic approach strengthens the identification of new therapeutic agents that target CSCs.
Antifungal bacterium against Colletotrichum coccodes causing black dot disease of potatoes and anthracnose of tomatoes was isolated from sewage sludge. The isolate showed a 99% sequence homology of partial 16S rRNA of Bacillus methylotrophicus CBMB205 and Bacillus amyloliquefaciens subsp. plantarum FZB42. The isolate was identified as Bacillus sp. SW29-2, using the neighbor-joining phylogenetic tree, BlastN sequence analysis, and morphological and cultural characteristics. Bacillus sp. SW29-2 is an aerobic, Gram-positive, endospore-forming bacterium, of which the morphological and physiological characteristics were the same as those of type strain B. lichniformis CBMB205, except for the cell growth of over 4% NaCl. The cell growth of the temperature and the initial pH of the medium was shown at $18-47^{\circ}C$ (opt. ca. $38^{\circ}C$) and 3-9 (opt. ca. 6.0), respectively. The inhibition size (diameter) of Bacillus sp. SW29-2 against four strains of C. coccodes ranged from 23 to 29 mm. Also, the isolate showed antifungal activity against penicillium rot-causing Penicillium expansum in apples. Thus far, any report on the antifungal activity of Baciilus spp. against C. coccodes has not been found. These results suggest that the Bacillus sp. SW29-2 isolate could be used as a possible biocontrol agent against C. coccodes, and further applied to other plant pathogenic fungi.
KSCE Journal of Civil and Environmental Engineering Research
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v.1
no.1
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pp.9-19
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1981
Using superplasticizer or the methods of autoclave curing is the most typical and practical of the ways to make high strength concrete. In this study the high strength concrete piles by way of centrifugal forming and hand compacting, are put to the freezing and thawing test after the process of autoclave curing, water curing after steam curing, or stand water curing. And the relation of such piles to durability, the rate of strength decrease, and the rate of weight variation is shown. The durability of those test piles throughout freezing and thawing test proves to be comparatively sufficient in the above-mentioned curings when the piles contain a proper quantity of air. But it is also acknowledged that without A.E agent, the high strength concrete piles of autoclave curing is higher than those of the other curing in strength, but lower in modulus of dynamic elasticity and their durability factor after the test is much lower.
Bone morphogenetic protein-2/4 (BMP-2/4) are members of Transforming Growth $Factor-{\beta}\;(TGF-{\beta})$ superfamily and they may differentiate the osteoprogenitor cell and induce formation of cartilage and bone in vivo. This study was performed to investigate the effects of bone morphogenetic protein-2/4 on the characteristics of rat periodontal ligament cells(RPDL) and rat calvaria cells(RCV). In the control group, the cells were cultured alone with Dulbeco's Modified Eagle's Medium contained with 20% fetal bovine serum, $100{\mu}/ml$ penicillin, $100{\mu}/ml$ streptomycin. In the experimental groups, recombinant human bone morphogenetic protein-2/4 (25ng, 100ng, 250ng/ml) were added into the above culture condition. And then each group was characterized by examing the cell proliferation at 1, 2, 3, 5, 7th day, the amount of total protein synthesis and alkaline phosphatase activity at 2, 5, 7th day. And also, the calcified nodule was examed. The results were as follows ; 1 . Both RCV and RPDL cells in both control and experimental groups proliferated during the entire experimental period, but there is no stastically significant difference according to the BMP-2/4 concentration. 2 . Amount of total protein synthesis of both cells in both groups was steadily increased until 5th day, but all experimental groups were significantly different from the control group at 7th day. 3. Alkaline phosphatase activity of both cells in both groups was increased during the entire experiment period. In RCV cells, the experimental group treated with 100ng/ml and 250ng/ml BMP-2/4 were significantly different from the control group at 7th day. In RPDL cells, the experimental group treated with 100ng/ml and 250ng/ml BMP-2/4 were significantly different from the control group at 5th day, and all experimental groups were significantly different from the control group at 7th day. 4. In the both of the cultured Rat Periodontal ligament and calvaria cell treated with BMP-2/4 to compared with control group, it revealed more rapid cell polarization, cell aggregation and hyperchromatic stained on HE agent, and even though only 1 day treated with BMP-2/4 both RPDL and RCV showed more rapid cell reaction than control group. More sensivitve cell reaction of RCV were observed than RPDL in this experiment. From the above results, we could conclude that BMP-2/4 influenced the induction, proliferation and differentiation of bone forming cells
Many Lactobacillus strains have been promoted as good probiotics for the prevention and treatment of diseases. We engineered recombinant Lactobacillus casei, producing biologically active canine granulocyte macrophage colony stimulating factor (cGM-CSF), and investigated its possibility as a good probiotic agent for dogs. Expression of the cGM-CSF protein in the recombinant Lactobacillus was confirmed by SDS-PAGE and Western blotting methods. For the in vivo study, 18 Beagle puppies of 7 weeks of age were divided into three groups; the control group was fed only on a regular diet and the two treatment groups were fed on a diet supplemented with either $1\times10^9$ colony forming units (CFU)/day of L. casei or L. casei expressing cGM-CSF protein for 7 weeks. Body weight was measured, and fecal and blood samples were collected from the dogs during the experiment for the measurement of hematology, fecal immunoglobulin (Ig)A and IgG, circulating IgA and IgG, and canine corona virus (CCV)-specific IgG. There were no differences in body weights among the groups, but monocyte counts in hematology and serum IgA were higher in the group receiving L. casei expressing cGM-CSF than in the other two groups. After the administration of CCV vaccine, CCV-specific IgG in serum increased more in the group supplemented with L. casei expressing cGM-CSF than the other two groups. This study shows that a dietary L. casei expressing cGM-CSF enhances specific immune functions at both the mucosal and systemic levels in puppies.
Journal of the Korea Organic Resources Recycling Association
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v.11
no.2
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pp.66-73
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2003
To investigate the availability of solidified wastes as resource, wastewater sludge, waste gypsum and fly ash were mixed and the results with various mixing ratios are as follows. Compressive strength turned out to be increasing as the amount of waste gypsum increases, keeps longer curing inhibition, and higher forming Pressure under the conditions of waste gypsum/sludge ratio 0.31-0.45, and 0.9kg cement as 15% and 1.2kg cement as 20% of total amount. Solidified agent under the fly ash/sludge ratio 0.45, 0.6, compressive strength seemed to be higher than standard one which means solidified wastes with these conditions could be applicable in real life. These results inform that concentrations of the leachate $Cr^{+6}$, Cu, Zn, Cd, Pb solidified matrix, containing low concentration of heavy metal, were cured with/without enough time it still will cause adverse effect on nature environment and application of heavy metal sequester must be needed to reuse industrial wastes from incineration plant solidified matrix. Total cost price, when considering manufacturing capability of the facilities for resourcerizing as 18,000ton was presented 678,664,000 won, as it were, manufacturing cost price was 37,704 won per ton. The results as above has shown that it's possible to use the mixture of waste gypsum/sludge, fly ash/sludge, cement, additions, and solidification matter as substitute of materials like brick, block, interlocking which has proper compressive strength of KS L 5201 and KS F 4004.
This paper was intended to evaluate the effect of the blending condition of pigments on the packing structure of coating color and the interaction between pigments and latices on the optical and interior properties of coated paper. It has been studied many ways to modify the coating structure to induce the interaction among coating components as followings ; 1) to use dispersant for pigment, 2) to control the charge density and the type of surface charge of latex, 3) to support the water retention by adding water retention agent or flow modifier. This paper was performed through the introduction of interaction between pigments which were two kinds of clays and one precipitated calcium carbonate(PCC) and ionic latices of which anionic and amphoteric respectively under the certain blending condition of pigments where their blending ratio of clays to calcium carbonate was 70pph to 30pph. The reason is that packing volume of pigments was highest in that region and thixotropical behavior appears in measuring rheology of coating color. We measured the properties of coating color, interaction with pigments and latex, and properties of coated paper and its printability. As a results, we could find out that amphoteric latex had a great influence on the interaction with pigments, especially clays, no matter what grade and also affected the coating structure significantly in case that this blending condition was 70(clays) to 30(PCC). It produced a powerful effect on the forming of bulky and smooth coating structure and in turn improved the printability of coated paper.
Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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2008.06a
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pp.546-546
/
2008
Copper (Cu) Chemical mechanical polishing (CMP) has been an essential process for Cu wifing of DRAM and NAND flash memory beyond 45nm. Copper has been employed as ideal material for interconnect and metal line due to the low resistivity and high resistant to electro-migration. Damascene process is currently used in conjunction with CMP in the fabrication of multi-level copper interconnects for advanced logic and memory devices. Cu CMP involves removal of material by the combination of chemical and mechanical action. Chemicals in slurry aid in material removal by modifying the surface film while abrasion between the particles, pad, and the modified film facilitates mechanical removal. In our research, we emphasized on the role of chemical effect of slurry on Cu CMP, especially on the effect of amine functional group on removal rate selectivity between Cu and Tantalum-nitride (TaN) film. We investigated the two different kinds of complexing agent both with amine functional group. On the one hand, Polyacrylamide as a polymer affected the stability of abrasive, viscosity of slurry and the corrosion current of copper film especially at high concentration. At higher concentration, the aggregation of abrasive particles was suppressed by the steric effect of PAM, thus showed higher fraction of small particle distribution. It also showed a fluctuation behavior of the viscosity of slurry at high shear rate due to transformation of polymer chain. Also, because of forming thick passivation layer on the surface of Cu film, the diffusion of oxidant to the Cu surface was inhibited; therefore, the corrosion current with 0.7wt% PAM was smaller than that without PAM. the polishing rate of Cu film slightly increased up to 0.3wt%, then decreased with increasing of PAM concentration. On the contrary, the polishing rate of TaN film was strongly suppressed and saturated with increasing of PAM concentration at 0.3wt%. We also studied the electrostatic interaction between abrasive particle and Cu/TaN film with different PAM concentration. On the other hand, amino-methyl-propanol (AMP) as a single molecule does not affect the stability, rheological and corrosion behavior of the slurry as the polymer PAM. The polishing behavior of TaN film and selectivity with AMP appeared the similar trend to the slurry with PAM. The polishing behavior of Cu film with AMP, however, was quite different with that of PAM. We assume this difference was originated from different compactness of surface passivation layer on the Cu film under the same concentration due to the different molecular weight of PAM and AMP.
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