Statistical optimization of the production medium was carried out in order to find an optimal medium composition in itaconic acid fermentation process. Itaconic acid utilized in the manufacture of various synthetic resins is a dicarboxylic acid biosynthesized by fungal cells of Aspergillus terreus in a branch of the TCA cycle via decarboxylation of cis-aconitate. Through OFAT (one factor at a time) experiments, six components (glucose, fructose, sucrose, soluble starch, soybean meal and cottonseed flour) were found to have significant effects on itaconic production among various carbon- and nitrogen-sources. Hence, using these six factors, interactive effects were investigated via fractional factorial design, showing that the initial concentrations of sucrose and cottonseed flour should be high for enhanced production of itaconic acid. Furthermore, through full factorial design (FFD) experiments, negative effects of $KH_2PO_4$ and $MgSO_4$ on itaconic acid biosynthesis were demonstrated, when excess amounts of the each component were initially added. Based on the FFD analysis, further statistical experiments were conducted along the steepest ascent path, followed by response surface method (RSM) in order to obtain optimal concentrations of the constituent nutrients. As a result, optimized concentrations of sucrose and cottonseed flour were found to be 90.4g/L and 53.8g/L respectively, with the corresponding production level of itaconic acid to be 4.36 g/L (about 7 fold higher productivity as compared to the previous production medium). From these experimental results, it was assumed that optimum ratio of the constituent carbon (sucrose) and nitrogen (cottonseed flour) sources was one of the most important factors for the enhanced production of itaconic acid.
Background : In acute lung injury, activated neutrophils play an important role in tissue damage. For neutrophils to participate in lung inflammation, chemotactic factors released from mononuclear phagocytes are needed to bring these cells to the local site of inflammation, with interleukin-8 (IL-8) being one of the most specific and important chemotactic factors for neutrophils. IL-8 also induces the expression of adhesion molecules and activates neutrophils to release various inflammatory mediators. Lipopolysaccharide(LPS) is one of the most important causes of adult respiratory distress syndrome and can cause release of many inflammatory cytokines including IL-8 leading to acute lung injury. But little is known about the regulatory mechanism of LPS-induced IL-8 gene expression in mononuclear phagocytes. Method : Human alveolar macrophages(HAM) and peripleral blood monocytes(PBMC) were isolated from healthy volunteers. Time and dose relationship of LPS-induced IL-8 mRNA expression was observed by Northern blot analysis. To evaluate the regulatory mechanism of LPS-induced IL-8 gene expression, pretreatment of actinomycin D(AD, $5{\mu}g/ml$) and cycloheximide(CHX, $5{\mu}g/ml$) was done and Northern blot analysis for IL-8 mRNA and ELISA for immunoreactive IL-8 protein in culture supernatant were performed. Results : 1) In HAM, dose and time dependent LPS-induced IL-8 mRNA expression was observed with peak mRNA level at 8 hours post-stimulation. 2) In PBMC, dose and time dependent LPS-induced IL-8 mRNA expression was also observed with peak mRNA level at 4 hours post-stimulation. 3) AD decreased expression of LPS-induced IL-8 gene expression at both mRNAand protein levels in both types of cells. 4) CHX decreased expression of LPS-induced IL-8 gene expression at protein level in both cell types but in HAM, superinduction of IL-8 mRNA was observed while decreased expression of IL-8 mRNA was observed in PBMC. Conclusion : Time and dose dependent LPS-induced IL-8 gene expression was observed in mononuclear phagocytes which is at least partly regulated pretranslationally. LPS-induced IL-8 mRNA expression in HAM needs no de novo protein synthesis and may be under the control of a labile repressor protein while de novo protein synthesis may be needed in PBMC.
Mineral carbonation is a technology in which carbonates are synthesized from minerals including serpentine and olivine, and industrial wastes such as slag and cement, of which all contain calcium or magnesium when reacted with carbon dioxide. This study aims to develop the mineral carbonation technology for commercialization, which can reduce environmental burden and process cost through the reduction of carbon dioxide using steel slag and the slag reuse after calcium extraction. Calcium extraction was conducted using NH4Cl solution for air-cooled slag and convert slag, and ${\geq}98%$ purity calcium carbonate was synthesized by reaction with calcium-extracted solution and carbon dioxide. And we conducted experimentally to minimize the quantity of by-product, the slag residue after calcium extraction, which has occupied large amount of weight ratio (about 80-90%) at the point of mineral carbonation process using slag. The slag residue was used to replace silica sand in the manufacture of cement panel, and physical properties including compressive strength and flexible strength of panel using the slag residue and normal cement panel, respectively, were analyzed. The calcium concentration in extraction solution was analyzed by inductively coupled plasma optical emission spectrometer (ICP-OES). Field-emission scanning electron microscope (FE-SEM) was also used to identify the surface morphology of calcium carbonate, and XRD was used to analyze the crystallinity and the quantitative analysis of calcium carbonate. In addition, the cement panel evaluation was carried out according to KS L ISO 679, and the compressive strength and flexural strength of the panels were measured.
The purpose of this animal study was to evaluate, by histological analysis, bone regeneration in rabbit maxillary sinuses with an anorganic bovine graft (Bio-Oss) and a ${\beta}-tricalcium$ phosphate (${\beta}-TCP$) grafting. Bilateral sinus augmentation procedures were performed in 12 adult male rabbits. Rectangular replaceable bony windows were made with a piezoelectric thin saw insert. In the Bio-Oss group, Bio-Oss was grafted and in the ${\beta}-TCP$ group, ${\beta}-TCP$ was grafted and covered by replaceable bony windows. The animals were sacrificed at 2, 4, and 8 weeks after the surgical procedure. The augmented sinuses were evaluated by histomorphometric analysis using hematoxylin-eosin, Masson trichrome, and tartrate-resistant acid phosphatase stains and also by immunohistochemical analysis of proliferating cell nuclear antigen (PCNA), type I collagen, and osteocalcin content. Histologically, new bone formation was found on the surface of Bio-Oss and ${\beta}-TCP$ particles from 2 weeks and continued to 8 weeks. Significant higher new bone formation was revealed in the ${\beta}-TCP$ group than in the Bio-Oss group at 8 weeks. The amount of graft materials was significantly decreased in the ${\beta}-TCP$ group and the number of osteoclasts was significantly increased in the ${\beta}-TCP$ group from 4 to 8 weeks. Immunoreactivity to PCNA was reduced at 8 weeks. The expression of type I collagen was significantly increased in the ${\beta}-TCP$ group at 2 weeks, but was significantly increased in the Bio-Oss group at 8 weeks. Immunoreactivity to osteocalcin was increased from 2 to 8 weeks. These histological results can help in the selection of graft materials for implants. Both Bio-Oss and ${\beta}-TCP$ are proven graft materials, however, these results indicate that ${\beta}-TCP$ showed better bone regeneration results in rabbit maxillary sinus augmentation.
Choi, Eun-Young;Lee, Jeong;Jeon, Min Ku;Lee, Sang-Kwon;Kim, Sung-Wook;Jeon, Sang-Chae;Lee, Ju Ho;Hur, Jin-Mok
Journal of the Korean Electrochemical Society
/
v.18
no.3
/
pp.121-129
/
2015
The platinum anode for the electrolytic reduction process is generally surrounded by a nonporous ceramic shroud with an open bottom to offer a path for $O_2$ gas produced on the anode surface and prevent the corrosion of the electrolytic reducer. However, the $O^{2-}$ ions generated from the cathode are transported only in a limited fashion through the open bottom of the anode shroud because the nonporous shroud hinders the transport of the $O^{2-}$ ions to the anode surface, which leads to a decrease in the current density and an increase in the operation time of the process. In the present study, we demonstrate the electrolytic reduction of 1 kg-uranium oxide ($UO_2$) using the porous shroud to investigate its long-term stability. The $UO_2$ with the size of 1~4mm and the density of $10.30{\sim}10.41g/cm^3$ was used for the cathode. The platinum and 5-layer STS mesh were used for the anode and its shroud, respectively. After the termination of the electrolytic reduction run in 1.5 wt.% $Li_2O-LiCl$ molten salt, it was revealed that the U metal was successfully converted from the $UO_2$ and the anode and its shroud were used without any significant damage.
Cancer is a complex disease heterogeneously composed of various types of cells including cancer stem-like cells responsible for relapse and chemoresistance in the tumor microenvironment. The conventional two-dimensional cell culture-based platform has critical limitations for representing the heterogeneity of cancer cells in the three-dimensional tumor niche in vivo. To overcome this insufficiency, three-dimensional cell culture methods in a scaffold-dependent or -free physical environment have been developed. In this study, we improved and simplified the HCT-8 colon cancer cell-based spheroid culture protocol and evaluated the relationship between cancer stemness and responses of chemosensitivity to 5- Fluorouracil (5-FU), a representative anticancer agent against colon cancer. Supplementation with defined growth factors in the medium and the culture dish of the regular surface with low attachment were required for the formation of constant-sized spheroids containing $CD44^+$ and $CD133^+$ colon cancer stem cells. The chemo-sensitivities of $CD44^+$ cancer stem cells in the spheroids were much lower than those of $CD44^-$ non-stem-like cancer cells, indicating that the chemoresistance to 5-FU is due to the stemness of colon cancer cells. Taken together, the inflammation and oncogenic gut environment-sensitive HCT-8 cell-based colon cancer spheroid culture and comparative evaluation using the simplified model would be an efficient and applicable way to estimate colon cancer stemness and pharmaceutical response to anticancer drugs in the realistic tumor niche.
Bakground : Complete resection by the surgery has been selected as the treatment of choice in lung cancer patients, but in cases of recurrence after excision or inoperable cases, the importance of anticancer chemotherapy has been emphasized. If one can select a set of the sensitive chemotherapeutic agents before anticancer chemotherapy, it will give more favourable results. Subrenal capsular assay has been recognized as a useful in-vivo chemosensitivity test of thoracic and abdominal tumors and it can be done in a short time for a rapid interpretation of tumor responsiveness to anticancer chemotherapeutic drugs. It has been reported that various kinds of cancer cells can be implantable to the kidney, but so far there is no comparative study of xenogeneic cell implantation on liver, spleen and kidney. The author implanted the human lung cancer cells under the capsule of S.D rat's liver, spleen and kidney respectively and compared the pattern of growth and histology. Material and Method: After incubation of human lung cancer cell line (SW-900 G IV) in RPMI 1640 (Leibovitz L-15 medium) culture media, 3${\times}$3${\times}$3 mm size fibrin clots which contain 108 cancer cells were made. Thereafter the fibrin clots were implanted at subcapsule area of liver, spleen and kidney of S.D. female rat. For immune suppression, cyclosporin-A (80 mg/Kg) was injected subcutaneously daily from post-implantation first day to sixth day. The body weight was measured at pre and post implantation periods. The growth pattern and the size of tumor mass were observed and the pathologic examination and serum tumor marker tests were performed. Result: Body weight increased in both of control and experimental groups. Serum Cyfra 21-1 was not detected. Serum levels of CEA and NSE revealed no significant change. The SCC-Ag increased significantly in implanted group. The growth rate of human lung cancer cells which was implanted on spleen was higher than on liver or kidney. The surface area, thickness, and volume of tumor mass were predominant at spleen. The success rates of implantation were 80% on kidney, 76.7% on spleen and 43.3% on liver. Pathologic examination of implanted tumors showed characteristic findings according to different organs. Tumors that were implanted on kidney grew in a round shape, small and regular pattern. In the spleen, tumors grew well and microscopic neovascularization and tumor thrombi were also found, but the growth pattern was irregular representing frequent daughter mass. Human lung cancer cells that were implanted in the liver, invaded to the liver parenchyme, and had low success rate of implantation. Microscopically, coagulation necrosis and myxoid fibrous lesion were observed. Conclusion: The success rate of implantation was highest in the kidney. And the mass revealed regular growth that could be measured easily. The SCC-Ag was presented earlier than CEA or Cyfra21-1. The Cyfra21-1 was not detected at early time after implantation. The best model for tumor implantation experiment for chemosensitivity test was subrenal capsular analysis than liver and spleen and the useful serum tumor marker in early period of implantation was the SCC-Ag.
Yu, Ri;Kwon, Young Sam;Oh, Tae-Ho;Kim, Tae-Hwan;Park, Sang-Joon
Journal of Veterinary Clinics
/
v.30
no.5
/
pp.339-345
/
2013
Mitogen-activated protein kinases (MAPKs) are a family of central signaling molecules that respond to numerous stimuli and are known to participate in processes of cell survival and death. However, it is not clear on data for cell-type specific activation of MAPKs in the progression of gastric ulcer. In the present study, we assessed how MAPKs localized at various cell types during the progression of gastric ulcer induced by ibuprofen. Gastric ulcer was induced by the repeated treatment of 200 mg/kg ibuprofen with 8 hrs interval in a day. Animals were sacrificed at 24 hrs, 48 hrs, and 72 hrs after oral treatment of ibuprofen and gastric tissues were subjected to immunohistochemical and immunoblotting evaluation. Immunoreactivity of phospho-extracellular signal-regulated kinase (p-ERK) was mainly expressed at the proliferating zone of gastric mucosa in control rats. But, these signals for p-ERK were highly shifted from cells of proliferating zone to parietal cells of the basal regions 24 hrs after treatment of ibuprofen. p-ERK signal was strongly expressed in epithelial cells adjacent to ulcer margin and new capillary and infiltrated inflammatory cells within granulation tissue of the ulcer base above 48 hrs after treatment of ibuprofen. While, phospho-c-Jun $NH_2$ terminal kinase (p-JNK) was mainly localized to the nuclei of the surface epithelial cells and the glandular epithelial cells in early gastric injury. Also, p-JNK was often observed as a scattered pattern in different regions of gastric mucosa with early gastric injury. Gradually, signal of p-JNK was strongly stained in infiltrated inflammatory cells and fibroblasts within severe ulcer base. Phospho-p38 (p-p38) MAPK was observed as scattered pattern within connective tissues of gastric mucosa. Especially, p-p38 MAPK showed strong signal in infiltrated macrophages within ulcer base. These results show that each MAPK has a specific role in various cell types during the progression of gastric ulcer.
The morphological, physical, and chemical properties of Sonjeong series derived from acidic crystalline rocks are presented. Also it deals with the genesis and classification of the Songjeong series. Morphologically these soils have brown to dark brown loam A horizons and yellowish red to red clay loam Bt horizons with moderate, medium subangular blocky structure and thin patchy clay cutans on the ped faces. C horizons are very deep, yellowish red to yellowish brown fine sandy loam or sandy loam with original rock structure. Physically distribution of particle size indicates that clay increases with depth up to argillic horizons but below the argillic horizons clay content decrease. The moisture holding capacity is fairly good in Songjeong soils. Chemically soil reaction is strongly to very strongly acid throughout the profile and content of organic matter is less than 1 per cent except A horizons. Cation exchange capacity ranges from 5 to 9 me/100g of soils and base saturation is less than 35 per cent throughout the profile. The natural fertility of Songjeong soils are usually low. It needs lime, organic matter, and heavy application of fertilizer for the crop land. These soils occur temperate and humid climate under coniferous, deciduous, and mixed forest vegetation. Songjeong soils are classified as Red-Yellow Soils. Characteristically Songjeong soils are similar to Red-Yellow Podzolic soils in the United States but lack of A2 horizons and are quite liket Red-Yellow Soils of the Japan. According to new classification system which is 7th approximation of USDA Songjeong soils can be classified as fine loamy, mesic family of Typic Hapludults and in the FAO/UNESCO project World Soil Map as Orthic Acrisols.
Kim, Dong-Myung;Kim, Seong-Ho;Lee, Jin-Man;Kim, Ji-Eun;Kang, Sun-Chul
Applied Biological Chemistry
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v.48
no.2
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pp.132-139
/
2005
The major obstacle in the popularization of Chungkookjang is the short shelf-life of $2{\sim}3$ months and some problems concerning storage including the growth of molds even in the products even within shelf-life. To solve these problems we conducted a research to improve its storage by using the vacuumed packaging and sanitary method through seed culture, innoculation and sterilization. For the optimization of storage time, temperature and sterilization temperature, we measured viable cell numbers of bacteria and fungi, amount of gas outbreak and contents of amino type nitrogen and monitored these experimental results by response surface methodology of SAS program, so that we could observe the quality changes of Chungkookjang during shelf-life. Especially fungi, which are the biggest troublemaker in Chungkookjang shelf-life, couldn't be detected from the generally and vacuum-packed samples; also, viable cell numbers were highly influenced by sterilization temperature and in vacuum-packed samples. In the case of vacuum-packed samples, amount of gas outbreak was highly influenced by sterilization temperature of its storage conditions and it was higher in generally packed samples as compared to vacuum-packed samples even at any storage conditions. The changes of pH in generally and vacuum-packed samples were highly influenced by the storage temperature. As the temperatures of storage and sterilization were higher and the storage time was longer, so the amount of gas outbreak was accordingly lower. These results showed that amino type nitrogen contents in generally and vacuum-packed samples were systematically influenced by the temperature, storage time and sterilization temperature. Also the result showed that the change of amino type nitrogen contents during storage was less in vacuum-packed samples than in general ones. Based on the above results, we can produce Chungkookjang products with extended shelf-life of as far as 6 months without any quality change using sanitary manufacturing method, vacuumed packaging condition, sterilization in $70^{\circ}C$ for 60 minutes and storage under $10^{\circ}C$ during shelf-life. According to this research, we have the possibility to greatly increase the goods value of Chungkookjang by developing the manufacture processing and packaging.
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