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Development of Immuno-Analytical System for Microbial Cells by using Dot-Blotter (Dot-Blotter 진공 포획방식에 의한 미생물세포 면역분석시스템의 개발)

  • 목락선;하연철;윤희주;백세환
    • KSBB Journal
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    • v.14 no.1
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    • pp.82-90
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    • 1999
  • In order to eventually fabricate an analytical system for infectious microorganisms, we synthesized major immunochemical components, utilized them for the construction of model system, and investigated an assay concept for bacterial whole cells. For the preparation of system components, a polyclonal antibody, against Salmonella thompson as model analyte, purified by immuno-affinity chromatography was used to chemically link to streptavidin or an enzyme, horseradish peroxidase(HRP). The antibody and streptavidin was modified with sulfosuccinimidyl 4-[N-maleimidomethyl]cyclohexane-1-carboxylate and N-succinimidyl-3-[2-pyridyldithio]propionate(subsequently activated by dithiotheritol), respectively. The modified components were reacted to synthesize antibody-streptavidin conjugates which were then purified on a two-layer chromatography column of diaminobiotin gel and Sephadex G-100. For antibody-HRP conjugates, HRP molecules were activated by $NalO_4$ oxidation and then coupled to immunoglobulin. After stabilizing with ($NaCNBH_3$, the conjugates were purified by size exclusion chromatography on Biogel A5M column. To devise a model system, such produced components were combined with a dot-blotter in which a nitrocellulose membrane($12{\mu}m$ pre size) with immobilized biotin was already located. The analyte (S. thompson cells) was reacted with the both antibody conjugates in a liquid phase, and the complexes formed were captured on the membrane surfaces by applying vacuum in the bottom compartment of the blotter to invoke biotin-streptavidin reaction. Under optimal conditions, the system enabled to identify the analytical concept for bacterial whole cells, and the lower limit of detection was approximately $1{\mu}g/m{\ell}$($10^5-10^6$ cells/m$m{\ell}$). The controlling factors were the concentrations of each antibody conjugate that caused agglutination in the presence of analyte as they increased.

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A Study on the Composition of Polar Lipid in Adlay (율무의 극성지질 조성에 관한 연구)

  • Han, Ji-Sook;Rhee, Sook-Hee;Cheigh, Hong-Sik
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.16 no.1
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    • pp.29-35
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    • 1987
  • The compositional characteristics of polar lipids in the brown and polished adlay were studied. Total lipids of brown and polished adlay were extracted, purified and fractionated into neutral and polar lipids by silicic acid column chromatography. The glycolipid(GL) and phospholipid(PL) of polar lipids were seperated by thin layer chromatography and quantitated by scanner. The fatty acid composition in polar lipids were determined by gas chromatographr. The contents of GL and PL in brown adlay were 5.67% and 1.83%, and their contents on polished adlay were 5.49% and 1.78%, respectively. Of the GL in the brown and polished adlay, monogalactosyl diglycerides+esterified steryl glycosides and digalactosyl diglycerides were the major components, but cerebrosides+steryl glycosides were also found as the minor component of GLs. Of the PLs, phosphatidyl choline and phosphatidyl ethanolamine were the major components. Phosphatidyl inositol, lysophosphatidyl choline and phosphatidyl serine were also present in the PLs. The major fatty acids of GLs were linoleic, oleic and palmitic acids. The fatty acid composition in the PLs were similar to the pattern in the GLs, but PLs contained the lower percentage of linoleic acids than the GLs.

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Effects of Soil Compaction upon the Vegetation Environment around the Trails in Pukhansan National Park (북한산(北漢山) 국립공원(國立公園)의 등산로(登山路) 주변(周邊)에서 답압(踏壓)이 식생환경(植生環境)에 미치는 영향(影響))

  • Suh, Min Hwan;Lee, Don Koo
    • Journal of Korean Society of Forest Science
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    • v.76 no.2
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    • pp.127-137
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    • 1987
  • This study was conducted to examine the effects of trampling on the changes in soil and vegetation around the trails. The study areas were established both on the trails and in the forested areas located in Ui and Jeongnung valleys at Pukhansan National Park. Soil core sampling, measurements of vegetations and counts of persons visited per hour were done during the period from April 6 to October 9, 1986. The physical properties of soils became better with increasing distances from the trail, and the best conditions were observed in the forested areas. The differences were not significant among the zones within the trailside. More number of individuals and basal area (or coverage) in woody plants were observed in the forested areas than at trailsides. For the herb layer, however, more number of individuals and coverage were shown at the trailside. In the Ui valley, the quantitative measures of herbaceous plants decreased as the bulk density and penetration resistance increased. In the Jeongnung valley, however, only bulk density was negatively related to the quantitative measures. The fact that there were poor relations between soil factors and quantitative measures of herbaceous plants in Jeongnung valley might be influenced by other factors rather than soil compaction. Thus, more research would be needed to understand the critical factors affecting the vegetational changes in recreation areas.

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Morphometric and Ultrastructural Change of Myelin-Associated Glycoprotein (MAG)-Immunoreactive Oligodendrocytes by Aging (노화에 의한 Myelin-Associated Glycoprotein (MAG) 면역반응 희소돌기아교세포의 형태계측학적 및 미세구조적 변화)

  • Cho, Ik-Hyun;Park, Chang-Hyun;Lee, Jong-Hwan;Bae, Chun-Sik;Ye, Sang-Kyu;Lee, Beob-Yi;Park, Seung-Hwa;Koh, Ki-Seok;Kim, Jin-Suk;Chang, Byung-Joon
    • Applied Microscopy
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    • v.36 no.2
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    • pp.119-130
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    • 2006
  • To investigate the role of myelin-associated glycoprotein (MAG) in the normal aging process, aging-related morphometric and ultrastructural analyses of the MAG-positive (MAG-(+)) oligodendrocytes were carried out in the cerebral cortex of the Sprague-Dawley rats. In the aged rats, the density of MAG-(+) oligodendrocytes was significantly decreased in the cortical layer (IV-VI) compared with that of the adult rats. However, the percentage of medium and dark types of oligodendrocytes was significantly increased by aging. In the aged rats, the mean nuclear area of the MAG-(-) oligodendrocytes was interestingly reduced compared with that of MAG-(+) oligodendrocytes. In addition, MAG immunoreactive products were markedly decreased in the medium-dark type of oligodendroglial cytoplasm and processes, and were scarcely localized in the dark type of oligodendrocytes of the aged rats. These results suggest that degeneration of oligodendrocytes-myelin system by aging is associated with down regulation of MAG, and that may contribute to further understanding of the biology of MAG in the oligodendrocytes-myelin system.

Histochemical and Fine Structural Visualization of the Epithelial Apoptosis in the Anuran Tadpole during the Tail Regression Stages (무미양서류 꼬리 퇴화과정중 피부 상피조직의 세포사에 관한 조직화학적 및 미세구조적 연구)

  • Lee, Hye-Won;Moon, Myung-Jin
    • Applied Microscopy
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    • v.39 no.2
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    • pp.107-115
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    • 2009
  • The programed cell death of the cutaneous epithelial tissue during tail regression stages in anuran tadpoles of the blackspotted frog, Rana nigromaculata were visualized by the histochemical and transmission electron microscopic techniques. Metamorphotic changes in the tail regression during the period of the Shumway stage number 31 to 33 are characterized by the disappearance of mucous layer and formation of compound epithelium through cutaneous thickening. Following the TUNEL (terminal deoxynucleotidyl transferase-mediated biotinylated d-uridine triphosphate nick end labeling) staining technique, the apoptotic cells were detected at the distal region of the tail skin initially, but they can be seen at the proximal region according to their following development. It has been also revealed that the number of the TUNEL-positive cells gradually increased from apical to basal direction of the epithelial layers during the tail regressing stages. Following the TEM observation, the early apoptotic cells shown in the epithelium demonstrated condensation and margination of the chromatin material at the nuclear periphery. Another epithelial apoptotic cells were shown nuclear fragmentation, membrane blebbing and cytoplasmic condensation. Following the process of the apoptotic degradation, well preserved organelles and nuclear fragments can be identified in the cytoplasm of lysosome-rich cells, however they soon reduced to lysosomal residual bodies through the progressive degradation.

Relationship between Degree of Grain Shedding and Histological Peculiarities of Abcission Region of Red Rice(Oryza sativa L.) Collected in Korea (국내 자생벼 계통의 탈립성과 이층조직의 특성)

  • 임원재;최광귀;진일두
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.35 no.3
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    • pp.239-247
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    • 1990
  • In order to investigate the degree of grain shedding of red rices collected in Korea, breaking tensile and breaking bending strengths of 269 varieties; 82 Korean and 100 foreign red rices and 87 Korean cultivars including 26 native varieties, 30 Japonica-Indica hybrids and 31 Japonica type varieties, were measured at harvest time. Also, histological characteristics of abscission region between spikelet and pedicel were observed. The breaking tensile and breaking bending strengths of both Korean and foreign red rices were weaker than those of Japonica-Indica hybrids which were known as easily shedding varieties in Korea. Abscission layers were observed in the majority of Korean red rices except for 5 varieties, and those of all varieties belonging to both long grain types and 'Sare's were cracked completely at harvest time, while those of round grain types were not cracked except one variety. The breaking tensile and breaking bending strengths of the tested varieties having cracked abscission layer were weaker than those of non-cracked ones. Both strengths of breaking tensile and breaking bending were positively correlated with diameter of supporting zone and thickness of sclerenchyma tissue surrounding central vascular tissue of suporting zone at 0.1% significant level, respectively.

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Cooking Properties of Rice with Pigmented Rice Bran Extract (유색미 미강 추출물 첨가가 밥의 취반 특성에 미치는 영향)

  • Kim, Joo-Hee;Nam, Seok-Hyun;Kim, Mi-Hyun;Sohn, Jae-Keun;Kang, Mi-Young
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.52 no.1
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    • pp.60-68
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    • 2007
  • This study was perform to examine the feasibility of cooking processing using the rice added the 70% ethanol extract of pigmented rice bran layer. Four rice samples, including normal rice, glutinous rice, pigmented-normal rice, and pigmented-glutinous rice were compared the properties of physico-chemical, texture, and sensory evaluation. Pigmented rice varieties had a higher amylose content, but shorter length in glucose chains than non-pigmented rice varieties. The enthalpy for gelatinization was found to increase in pigmented rice, which need more energy for gelatinization of starch in cooking. The hydrolysis rate by glucoamylase in rice added pigmented bran extract was higher than pigmented rice. Rice with pigmented bran extract had higher glutamine content, but lower asparagine content and no difference in fatty acid composition, which affect palatability. Cooked rice added pigmented bran extract was less retrograded than pigmented rice during the storage period. Moreover, cooked rice added pigmented bran extract was more acceptable in sensory evaluation. Based on the results, the use of rice added pigmented bran extract instead of pigmented rice in grain processed food have advantageous effects in palatability of polished rice and phytochemicals of pigmented non-polished rice. This study will help develop new health-promoting rice products.

Prediction of Homogenization Efficiency using Response Surface Methodology (반응표면분석을 활용한 균질 효율 예측)

  • Kang, Ho Jin;Kang, Shin Ho;Shin, Yong Kook
    • Journal of Dairy Science and Biotechnology
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    • v.35 no.3
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    • pp.202-207
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    • 2017
  • The objective of this study was to analyze the effects of homogenization, storage temperature, and storage period on the creaming of milk fat and changes in fat contents in the upper and lower layers and to predict the conditions for optimal homogenization efficiency using response surface methodology (RSM). The homogenization pressure, storage temperature, and storage period were set as independent variables of RSM, and the dependent variables were creaming, US Public Health Service (USPHS) code, and volume weighted mean diameter ($D_{4,3}$) in the upper and lower layers. Based on the results of RSM and regression analysis, the correlation coefficient ($R^2$) between experimental data and predicted values by RSM for homogenized milk was estimated to be more than 0.8. The RSM analysis indicated that optimal homogenization pressures of 14 MPa or more and 17 MPa or more were required to maintain the creaming layer of 3 mm or less during the storage for 15 days at $10^{\circ}C$ and $20^{\circ}C$, respectively. To keep the USPHS code at less than 10% for 15 days at $10^{\circ}C$ and $20^{\circ}C$, milk should be homogenized with a pressure of 16.8 MPa or more and 17 MPa or more, respectively.

Histopathological response of flounder, Paralichthys olivaceus, treated with formlin and neutral-formalin (포르말린 및 중성 포르말린 약욕 넙치(Paralichthys olivaceus)의 병리조직학적 반응)

  • Cho, Jae-Kwon;Chun, Min-Nam;Yang, Han-Choon
    • Journal of fish pathology
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    • v.10 no.1
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    • pp.53-63
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    • 1997
  • In this study, poikilocyte rate and histopathological observation of flounder, Paralichthys olivaceus, treated with formalin, neutral-formalin(0, 50, 100, 150, 200, 300 ppm) at two different temperatures (15 and $25^{\circ}C$) were conducted to determine the time need for recovery. In all test chamber, formalin was more toxic than neutral-formalin and moderate lesions were not observed at low concentration of formalin and neutral-formalin(50~100 ppm). As expectedly, time need for recovery of gill and kidney tissues was lengthening as increasing chemical concentration and temperature. Treatments caused edema, winding of secondary gill lamella, seperation of epithelial layer and thrombosis of secondary gill lamella in the gill; edema, hydropic and hyaline droplet degeneration in the kidney. Representatively, recovery period of fish gill that had been exposed to 300 ppm formalin and neutral-formalin was about 120 and 72 hr($25^{\circ}C$). 72 and 48 hr($15^{\circ}C$). Recovery period of kidney was about 72 and 48 hr($25^{\circ}C$), 48 and 48 hr($15^{\circ}C$) respectively. Maximum value of poikilocyte rate(27.84%) was shown in formalin 300 ppm treated fish at $25^{\circ}C$. Tendency of decreasing poikilocyte rate was similiar to recovery of tissues in treated fish.

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Si and Mg doped Hydroxyapatite Film Formation by Plasma Electrolytic Oxidation

  • Park, Seon-Yeong;Choe, Han-Cheol
    • Proceedings of the Korean Institute of Surface Engineering Conference
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    • 2016.11a
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    • pp.195-195
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    • 2016
  • Titanium and its alloys are widely used as implants in orthopedics, dentistry and cardiology due to their outstanding properties, such as high strength, high level of hemocompatibility and enhanced biocompatibility. Hence, recent works showed that the synthesis of new Ti-based alloys for implant application involves more biocompatible metallic alloying element, such as, Nb, Hf, Zr and Mo. In particular, Nb and Hf are one of the most effective Ti ${\beta}-stabilizer$ and reducing the elastic modulus. Plasma electrolyte oxidation (PEO) is known as excellent method in the biocompatibility of biomaterial due to quickly coating time and controlled coating condition. The anodized oxide layer and diameter modulation of Ti alloys can be obtained function of improvement of cell adhesion. Silicon (Si) and magnesium (Mg) has a beneficial effect on bone. Si in particular has been found to be essential for normal bone and cartilage growth and development. In vitro studies have shown that Mg plays very important roles in essential for normal growth and metabolism of skeletal tissue in vertebrates and can be detected as minor constituents in teeth and bone. The aim of this study is to research Si and Mg doped hydroxyapatite film formation by plasma electrolytic oxidation. Ti-29Nb-xHf (x= 0, 3, 7 and 15wt%, mass fraction) alloys were prepared Ti-29Nb-xHf alloys of containing Hf up from 0 wt% to 15 wt% were melted by using a vacuum furnace. Ti-29Nb-xHf alloys were homogenized for 2 hr at $1050^{\circ}C$. Each alloy was anodized in solution containing typically 0.15 M calcium acetate monohydrate + 0.02 M calcium glycerophosphate at room temperature. A direct current power source was used for the process of anodization. Anodized alloys was prepared using 270V~300V anodization voltage at room. A Si and Mg coating was produced by RF-magnetron sputtering system. RF power of 100W was applied to the target for 1h at room temperature. The microstructure, phase and composition of Si and Mg coated oxide surface of Ti-29Nb-xHf alloys were examined by FE-SEM, EDS, and XRD.

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