• Title/Summary/Keyword: Plate Test

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Mass Screening of Lovastatin High-yielding Mutants through Statistical Optimization of Sporulation Medium and Application of Miniaturized Fungal Cell Cultures (Lovastatin 고생산성 변이주의 신속 선별을 위해 통계적 방법을 적용한 Sporulation 배지 개발 및 Miniature 배양 방법 개발)

  • Ahn, Hyun-Jung;Jeong, Yong-Seob;Kim, Pyeung-Hyeun;Chun, Gie-Taek
    • KSBB Journal
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    • v.22 no.5
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    • pp.297-304
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    • 2007
  • For large and rapid screening of high-yielding mutants of lovastatin produced by filamentous fungal cells of Aspergillus terreus, one of the most important stage is to test as large amounts of mutated strains as possible. For this purpose, we intended to develop a miniaturized cultivation method using $7m{\ell}$ culture tube instead of traditional $250m{\ell}$ flask (working volume $50m{\ell}$). For obtaining large amounts of conidiospores to be used as inoculums for miniaturized cultures, 4 components i.e., glucose, sucrose, yeast extract and $KH_2PO_4$ were intensively investigated, which had been observed to show positive effect on enhancement of spore production through Plackett-Burman design experimet. When optimum concentrations of these components that were determined through application of response surface method (RSM) based on central composite design (CCD) were used, maximum spore numbers amounting to $1.9\times10^{10}$ spores/plate were obtained, resulting in approximately 190 fold increase as compared to the commonly used PDA sporulation medium. Using the miniaturized cultures, intensive strain development programs were carried out for screening of lovastatin high-yielding as well as highly reproducible mutants. It was observed that, for maximum production of lovastatin, the producers should be activated through 'PaB' adaptation process during the early solid culture stage. In addition, they should be proliferated in condensed filamentous forms in miniaturized growth cultures, so that optimum amounts of highly active cells could be transferred to the production culture-tube as reproducible inoculums. Under these highly controlled fermentation conditions, compact-pelleted morphology of optimum size (less than 1 mm in diameter) was successfully induced in the miniaturized production cultures, which proved essential for maximal utilization of the producers' physiology leading to significantly enhanced production of lovastatin. As a result of continuous screening in the miniaturized cultures, lovastatin production levels of the 81% of the daughter cells derived from the high-yielding producers turned out to be in the range of 80%$\sim$120% of the lovastatin production level of the parallel flask cultures. These results demonstrate that the miniaturized cultivation method developed in this study is efficient high throughput system for large and rapid screening of highly stable and productive strains.

Establishment and Characterization of Bone Marrow Mesenchymal Stromal/Stem Cells (MSCs) Derived from ${\alpha}$-1,3-Galactosyltransferase Knock Out(GalT KO) Pig (${\alpha}$-1,3-Galactosyltransferase Knock Out(GalT KO) 돼지유래 골수 중간엽 줄기세포의 특성 규명)

  • Ock, Sun-A;Oh, Keon Bong;Hwang, Seongsoo;Im, Seoki;Kim, Youngim;Park, Jin-Ki
    • Journal of Embryo Transfer
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    • v.28 no.3
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    • pp.281-287
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    • 2013
  • A major barrier to progress in pig to primate organ transplantation or cell therapy is the presence of terminal ${\alpha}$-1,3-galactosyl epitopes on the surface of pig cells. Therefore, the purpose of this experiment was to establish and cha- racterize mesenchymal stromal/stem cells (MSCs) derived from ${\alpha}$-1,3-galactosyltransferase (GalT) knock out (GalT KO) pig to confirm their potential for cell therapy. Bone marrow (BM)-MSCs from GalT KO pig of 1 month old were isolated by Ficoll-Paque PLUS gradient and cultured with A-DMEM + 10% FBS on plastic dishes in 5% $CO_2$ incubator at 38.5. GalT KO BM-MSCs were analyzed for the expression of CD markers ($CD45^-$, $29^+$, $90^+$ and $105^+$) and in vitro differentiation ability (adiopogenesis and osteogenesis). Further, cell proliferation capacity and cell aging of GalT KO BM-MSCs were compared to Wild BM-MSCs by BrdU incorporation assay (Roche, Germany) using ELISA at intervals of two days for 7 days. Finally, the cell size was also evaluated in GalT KO and Wild BM-MSCs. Statistical analysis was performed by T-test (P<0.05). GalT KO BM-MSCs showed fibroblast-like cell morphology on plastic culture dish at passage 1 and exhibited $CD45^-$, $29^+$, $90^+$ and $105^+$ expression profile. Follow in ginduction in StemPro adipogenesis and osteogenesis media for 3 weeks, GalT KO BM-MSCs were differentiated into adipocytes, as demonstrated by Oilred Ostaining of lipid vacuoles and osteocytes, as confirmed by Alizarinred Sstaining of mineral dispositions, respectively. BrdU incorporation assay showed a significant decrease in cell proliferation capacity of GalT KO BM-MSCs compared to Wild BM-MSCs from 3 day, when they were seeded at $1{\times}10^3$ cells/well in 96-well plate. Passage 3 GalT KO and Wild BM-MSCs at 80% confluence in culture dish were allowed to form single cells to calculate cell size. The results showed that GalT KO BM-MSCs($15.0{\pm}0.4{\mu}m$) had a little larger cell size than Wild BM-MSCs ($13.5{\pm}0.3{\mu}m$). From the above findings, it is summarized that GalT KO BM-MSCs possessed similar biological properties with Wild BM-MSCs, but exhibited a weak cell proliferation ability and resistance to cell aging. Therefore, GalT KO BM-MSCs might form a good source for cell therapy after due consideration to low proliferation potency in vitro.

Effects of Gamma Irradiation on Color Changes and Biological Activities of Ethanol Extract of a Mechanically Pressed Juice of Bokbunja (Rubus coreanus Miq.) (감마선 조사에 의한 복분자 착즙액 에탄올 추출물의 색상 및 생리활성 변화)

  • Kim, Hee-Jung;Jo, Cheor-Un;Kim, Hyun-Joo;Shin, Dong-Hwa;Son, Jun-Ho;Byun, Myung-Woo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.35 no.3
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    • pp.271-277
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    • 2006
  • A mechanically pressed juice of Bokbunja (Rubus coreanus Miq.) extract was prepared using 70% ethanol solution. The extract was subjected to gamma-irradiation treatment (20 kGy) and investigated for its change of color and biological activities. Hunter $L^*$ values of the irradiated Bokbunja extract were increased in comparison with the non-irradiated extracts, and the $a^*\;and\;b^*$values decreased by the irradiation treatment. The content of the total phenolic compounds in the non-irradiated and irradiated extracts were 58.4 and 56.5 mg/g, respectively. The 1,1-diphenyl-2-picrylhydrazyl(DPPH) radical scavenging activities of non-irradiated and irradiated extracts at a 250 ppm level were 80% and 79%, respectively. Lipid oxidation was retarded by addition of Bokbunja powder. Nitrite scavenging activity was the highest in the Bokbunja powder at pH 1.2 and the effect was not changed by irradiation. The Bokbunja powder showed antimicrobial activity against Salmonella Typhimurium and Bacillus cereus. However, irradiation of Bokbunja did not affect any physiological functions (p>0.05). A Salmonella mutagenicity assay indicated that the irradiated Bokbunja extract did not show any mutagenicity. Therefore, Bokbunja extract could be used in various applications as a functional material, such as ingredients of food and cosmetic, compositions with functions.

Dynamic Characteristic Analysis Procedure of Helicopter-mounted Electronic Equipment (헬기 탑재용 전자장비의 동특성 분석 절차)

  • Lee, Jong-Hak;Kwon, Byunghyun;Park, No-Cheol;Park, Young-Pil
    • Transactions of the Korean Society for Noise and Vibration Engineering
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    • v.23 no.8
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    • pp.759-769
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    • 2013
  • Electronic equipment has been applied to virtually every area associated with commercial, industrial, and military applications. Specifically, electronics have been incorporated into avionics components installed in aircraft. This equipment is exposed to dynamic loads such as vibration, shock, and acceleration. Especially, avionics components installed in a helicopter are subjected to simultaneous sine and random base excitations. These are denoted as sine on random vibrations according to MIL-STD-810F, Method 514.5. In the past, isolators have been applied to avionics components to reduce vibration and shock. However, an isolator applied to an avionics component installed in a helicopter can amplify the vibration magnitude, and damage the chassis, circuit card assembly, and the isolator itself via resonance at low-frequency sinusoidal vibrations. The objective of this study is to investigate the dynamic characteristics of an avionics component installed in a helicopter and the structural dynamic modification of its tray plate without an isolator using both a finite element analysis and experiments. The structure is optimized by dynamic loads that are selected by comparing the vibration, shock, and acceleration loads using vibration and shock response spectra. A finite element model(FEM) was constructed using a simplified geometry and valid element types that reflect the dynamic characteristics. The FEM was verified by an experimental modal analysis. Design parameters were extracted and selected to modify the structural dynamics using topology optimization, and design of experiments(DOE). A prototype of a modified model was constructed and its feasibility was evaluated using an FEM and a performance test.

DETECTION OF METHICILLIN OR VANCOMYCIN-RESISTANT STAPHYLOCOCCUS AUREUS FROM DENTAL HOSPITAL (치과병원 진료실 내에서 메티실린 또는 반코마이신 저항성 Staphylococcus aureus의 검출)

  • Min, Jung-Hee;Park, Soon-Nang;Hwang, Ho-Keel;Min, Jung-Beum;Kim, Hwa-Sook;Kook, Joong-ki
    • Restorative Dentistry and Endodontics
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    • v.32 no.2
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    • pp.102-110
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    • 2007
  • The purpose of this study was to obtain the basic information for the improvement of dental environment by investigating the presence of methicillin- or vancomycin-resistant Staphylococcus aureus (MRSA or VRSA) isolated from dental health care workers (DHCWs) and environment of the Chosun University Dental Hospital (CUDH) and a private dental clinic (control group). Staphylococcus aureus (S. aureus) was isolated from anterior nares of 42 DHCWS and 38 sites, unit chairss, x-ray devices, computers, etc., at 10 departments of the CUDH and 20 DHCWs and 11 sites at the private dental clinic. S. aureus was isolated on mannitol salt agar plate and confirmed by PCR with S. aureus species-specific primer. Antimicrobial susceptibility test of clinical isolates of S. aureus against several antibiotics including methicillin (oxacillin) was performed by investigating minimum inhibitory concentration (MIC) using broth microdilution assay. In addition, PCR was performed to detect the methicillin- or vancomycin-resistant gene. The data showed that one strain of S. aureus was isolated from DHCWs of the CUDH and three strains of S. aureus was isolated from 3 samples of the private dental clinic, respectively. All of the isolates from the CUDH and the private dental clinic had resistance to penicillin G, amoxicillin and vancomycin and susceptibility to oxacillin and ciprofloxacin. The S. aureus strains were already obtained the resistance to penicillin G and amoxicillin. These results suggest that two dental clinics were under relatively safe environment.

Comparison of Quality Characteristics in Dry-Cured Ham at Initial Stage of Distribution (유통초기단계의 건조- 숙성햄의 품질특성 비교)

  • Jin, Sang-Keun;Kim, Il-Suk;Yang, Mi-Ra;Hur, In-Chul;Kim, Dae-Seung;Kang, Suk-Nam
    • Journal of Animal Science and Technology
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    • v.53 no.4
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    • pp.377-385
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    • 2011
  • This study was carried out to investigate quality characteristics of dry-cured hams in the domestic market. Two kinds of dry-cured hams were processed with pork leg (T1) and pork neck (T2). Five 24-month dry-cured legs (T1, 5.3-6.1 kg) and twenty 4-month dry-cured pork necks (T2, 1.5-1.9 kg) were used in this experiment. They were stored at $4^{\circ}C$ chilling room and quality characteristics were investigated. Moisture content, water activity, cholesterol content, CIE $L^*$, VBN, total plate counts and lactic acid bacteria of T1 were significantly higher (p<0.05) than those of T2, whereas fat content, ${NO_2}^-$, CIE $a^*$, TBARS and Warner-Bratzler shear force values of T2 were significantly higher (p<0.05) than those of T1. In fatty acid compositions, palmitoleic acid and stearic acid in T2 were significantly higher (p<0.05) than T1, however, oleic and linoleic acid in T2 was significantly lower (p<0.05) than T1. In free amino acids, the total content and individual content of asparagin, leucine and phenylalanine in T1 were higher than those of T2 (p<0.05). The aroma score of T2 was higher than that of T1 in sensory evaluation (p<0.05). In conclusion, two kinds of dry-cured hams were different in their final characteristics and could enhance the consumer's appeal of pork meat in Korean market.

The Electrochemical Chlorination for Marine Plankton Community Disinfection (해양 플랑크톤 군집의 전기분해 염소소독 효과)

  • Kang, Jung-Hoon;Shin, Kyoung-Soon;Hyun, Bong-Gil;Jang, Min-Chul;Kim, Eun-Chan;Chang, Man
    • Journal of the Korean Society for Marine Environment & Energy
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    • v.10 no.3
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    • pp.127-137
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    • 2007
  • To confirm whether or not the Electrochemical Disinfection System (EDS) meet with the D-2 regulation established by IMO (International Maritime Organization), the biological treatment efficacy of the EDS was assessed using three groups of natural marine plankton (bacteria, $10-50\;{\mu}m$ and $>50\;{\mu}m$ sized organisms). Influent water was passed through the EDS under the flow velocity ($23.8\;m^3/hr$) and test design was consisted of control (no treatment) and experimental (10 ppm and 30 ppm) condition for total residual chlorine (TRC). And the biological condition of the influent water followed the standards established by the guidelines for the approval of ballast water management systems. The disinfection efficacy of the $10-50\;{\mu}m$ sized organisms (phytoplankton) was assessed by three kinds of measurements using photomicroscope, epifluorescence microscope and fluorometer (fumer Designs 10-AU). After being passed through the EDS, all motile phytoplankton lost their motility under photomicroscope, the colour of chlorophyll fluorescence fumed from red into green under epifluorescence, and the high chlorophyll fluorescence (Expt. 1: 6.95, Expt. 2: 7.11) detected by fluorometer decreased into value not detected. These results indicated phytoplankton community was totally killed after electrochemical disinfection treatment. Survivorship of the larger organisms than $50\;{\mu}m$ was determined based on the appendage's movement under a stereomicroscope. Natural assemblage collected from ambient seawater was killed shortly after being passed through the EDS, whereas some Artemia remained alive. However, no live Artemia was found after 24 hour further exposure to each TRC concentration (10 and 30 ppm) under darkness. After electrochemical treatment, the target bacteria such as aerobes, coliform and Escherichia coli were completely killed on the basis of CFU (colony forming unit) on Petrifilm plate ($3\;M^{TM}$) after 48 hr incubation. Moreover, no regrowth was found in the three groups of plankton during five days under additional exposure to the treated water. These results indicated that the disinfection efficiency of the EDS on the three groups of plankton satisfy D-2 regulation.

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Optimization of a Medium for the Production of Cellulase by Bacillus subtilis NC1 Using Response Surface Methodology (반응 표면 분석법을 사용한 Bacillus subtilis NC1 유래 cellulase 생산 배지 최적화)

  • Yang, Hee-Jong;Park, Chang-Su;Yang, Ho-Yeon;Jeong, Su-Ji;Jeong, Seong-Yeop;Jeong, Do-Youn;Kang, Dae-Ook;Moon, Ja-Young;Choi, Nack-Shick
    • Journal of Life Science
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    • v.25 no.6
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    • pp.680-685
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    • 2015
  • Previously, cellulase and xylanase producing microorganism, Bacillus subtilis NC1, was isolated from soil. Based on the 16S rRNA gene sequence and API 50 CHL test the strain was identified as Bacillus subtilis, and named as B. subtilis NC1. We cloned and sequenced the genes for cellulase and xylanase. Plus, the deduced amino acid sequences from the genes of cellulase and xylanase were determined and were also identified as glycosyl hydrolases family (GH) 5 and 30, respectively. In this study to optimize the medium parameters for cellulase production by B. subtilis NC1 the RSM (response surface methodology) based on CCD (central composite design) model was performed. Three factors, tryptone, yeast extract, and NaCl, for N or C source were investigated. The cellulase activity was measured with a carboxylmethyl cellulose (CMC) plate and the 3,5-dinitrosalicylic acid (DNS) methods. The coefficient of determination (R2) for the model was 0.960, and the probability value (p=0.0001) of the regression model was highly significant. Based on the RSM, the optimum conditions for cellulase production by B. subtilis NC1 were predicted to be tryptone of 2.5%, yeast extract of 0.5%, and NaCl of 1.0%. Through the model verification, cellulase activity of Bacillus subtilis NC1 increased from 0.5 to 0.62 U/ml (24%) compared to the original medium.

Quality Changes and Shelf-life of Seasoned Pork with Soy Sauce or Kochujang during Chilled Storage (간장과 고추장 양념 돈육의 냉장 중 품질 변화와 저장 수명)

  • 최원선;이근택
    • Food Science of Animal Resources
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    • v.22 no.3
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    • pp.240-246
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    • 2002
  • The seasoned pork with soy sauce(SS) or Kochujang(SK) was manufactured using hind and fore leg as main raw material and the quality characteristics and shelf-life were investigated during storage at 5$\^{C}$ and 10$\^{C}$ after having packaged with air. The initial total aerobic plate counts(APC) of SS and SK were 5.24 and 5.75 log10 CFU/㎠, respectively. APC exceeded 7 log10CFU/cm2 after 10 days at 5$\^{C}$ and 6 days at 10$\^{C}$ far SS, and after 6 days at 5$\^{C}$ and 4 days at 10$\^{C}$ for SK, respectively. In the sensory test, the SS samples stored at 5$\^{C}$ and 10$\^{C}$ were assessed as lower than 3.0 points, the criterium of consumer acceptability, after 10 and 8 days, respectively. In the case of SK samples, the point of this time was at day 10 and 6, respectively when they were stored at 5$\^{C}$ and 10$\^{C}$. As storage time extended, pH and water content tended to decrease. Contrarily, the increase of TBA and VBN values was observed during storage and this was more pronounced at 10$\^{C}$ than at 5$\^{C}$ . The oxygen concentration in the package was 19.8 and 19.9% each for 55 and SK samples at the beginning but it was gradually decreased thereafter. On the other hand, the concentration of carbon dioxide was increased with the extension of storage. Based on the above results, the shelf-lives of seasoned pork were estimated to be 8 days for SS and SK stored at 5$\^{C}$ and 6 days for SS and 4 days for SK stored at 10$\^{C}$.

Experimental Studies on the Properties of Epoxy Resin Mortars (에폭시 수지 모르터의 특성에 관한 실험적 연구)

  • 연규석;강신업
    • Magazine of the Korean Society of Agricultural Engineers
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    • v.26 no.1
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    • pp.52-72
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    • 1984
  • This study was performed to obtain the basic data which can be applied to the use of epoxy resin mortars. The data was based on the properties of epoxy resin mortars depending upon various mixing ratios to compare those of cement mortar. The resin which was used at this experiment was Epi-Bis type epoxy resin which is extensively being used as concrete structures. In the case of epoxy resin mortar, mixing ratios of resin to fine aggregate were 1: 2, 1: 4, 1: 6, 1: 8, 1:10, 1 :12 and 1:14, but the ratio of cement to fine aggregate in cement mortar was 1 : 2.5. The results obtained are summarized as follows; 1.When the mixing ratio was 1: 6, the highest density was 2.01 g/cm$^3$, being lower than 2.13 g/cm$^3$ of that of cement mortar. 2.According to the water absorption and water permeability test, the watertightness was shown very high at the mixing ratios of 1: 2, 1: 4 and 1: 6. But then the mixing ratio was less than 1 : 6, the watertightness considerably decreased. By this result, it was regarded that optimum mixing ratio of epoxy resin mortar for watertight structures should be richer mixing ratio than 1: 6. 3.The hardening shrinkage was large as the mixing ratio became leaner, but the values were remarkably small as compared with cement mortar. And the influence of dryness and moisture was exerted little at richer mixing ratio than 1: 6, but its effect was obvious at the lean mixing ratio, 1: 8, 1:10,1:12 and 1:14. It was confirmed that the optimum mixing ratio for concrete structures which would be influenced by the repeated dryness and moisture should be rich mixing ratio higher than 1: 6. 4.The compressive, bending and splitting tensile strenghs were observed very high, even the value at the mixing ratio of 1:14 was higher than that of cement mortar. It showed that epoxy resin mortar especially was to have high strength in bending and splitting tensile strength. Also, the initial strength within 24 hours gave rise to high value. Thus it was clear that epoxy resin was rapid hardening material. The multiple regression equations of strength were computed depending on a function of mixing ratios and curing times. 5.The elastic moduli derived from the compressive stress-strain curve were slightly smaller than the value of cement mortar, and the toughness of epoxy resin mortar was larger than that of cement mortar. 6.The impact resistance was strong compared with cement mortar at all mixing ratios. Especially, bending impact strength by the square pillar specimens was higher than the impact resistance of flat specimens or cylinderic specimens. 7.The Brinell hardness was relatively larger than that of cement mortar, but it gradually decreased with the decline of mixing ratio, and Brinell hardness at mixing ratio of 1 :14 was much the same as cement mortar. 8.The abrasion rate of epoxy resin mortar at all mixing ratio, when Losangeles abation testing machine revolved 500 times, was very low. Even mixing ratio of 1 :14 was no more than 31.41%, which was less than critical abrasion rate 40% of coarse aggregate for cement concrete. Consequently, the abrasion rate of epoxy resin mortar was superior to cement mortar, and the relation between abrasion rate and Brinell hardness was highly significant as exponential curve. 9.The highest bond strength of epoxy resin mortar was 12.9 kg/cm$^2$ at the mixing ratio of 1:2. The failure of bonded flat steel specimens occurred on the part of epoxy resin mortar at the mixing ratio of 1: 2 and 1: 4, and that of bonded cement concrete specimens was fond on the part of combained concrete at the mixing ratio of 1 : 2 ,1: 4 and 1: 6. It was confirmed that the optimum mixing ratio for bonding of steel plate, and of cement concrete should be rich mixing ratio above 1 : 4 and 1 : 6 respectively. 10.The variations of color tone by heating began to take place at about 60˚C, and the ultimate change occurred at 120˚C. The compressive, bending and splitting tensile strengths increased with rising temperature up to 80˚ C, but these rapidly decreased when temperature was above 800 C. Accordingly, it was evident that the resistance temperature of epoxy resin mortar was about 80˚C which was generally considered lower than that of the other concrete materials. But it is likely that there is no problem in epoxy resin mortar when used for unnecessary materials of high temperature resistance. The multiple regression equations of strength were computed depending on a function of mixing ratios and heating temperatures. 11.The susceptibility to chemical attack of cement mortar was easily affected by inorganic and organic acid. and that of epoxy resin mortar with mixing ratio of 1: 4 was of great resistance. On the other hand, when mixing ratio was lower than 1 : 8 epoxy resin mortar had very poor resistance, especially being poor resistant to organicacid. Therefore, for the structures requiring chemical resistance optimum mixing of epoxy resin mortar should be rich mixing ratio higher than 1: 4.

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