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Expression of CD40, CD86, and HLA-DR in CD1c+ Myeloid Dendritic Cells Isolated from Peripheral Blood in Primary Adenocarcinoma of Lung (원발성 폐선암환자의 말초혈액에서 분리한 CD1c+ 골수성 수지상 세포에서의 CD40, CD86 및 HLA-DR의 발현)

  • Kang, Moon-Chul;Kang, Chang-Hyun;Kim, Young-Tae;Kim, Joo-Hyun
    • Journal of Chest Surgery
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    • v.43 no.5
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    • pp.499-505
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    • 2010
  • Background: There have been several reports using animal experiments that CD1-restricted T-cells have a key role in tumor immunity. To address this issue, we studied the expression of markers for CD1c+ myeloid dendritic cells (DCs) isolated from peripheral blood in the clinical setting. Material and Method: A total of 24 patients with radiologically suspected or histologically confirmed lung cancer who underwent pulmonary resection were enrolled in this study. The patients were divided according to histology findings into three groups: primary adenocarcinoma of lung (PACL), primary squamous cell carcinoma of lung (PSqCL) and benign lung disease (BLD). We obtained 20 mL of peripheral venous blood from patients using heparin-coated syringes. Using flow-cytometry after labeling with monoclonal antibodies, data acquisition and analysis were done. Result: The ratio of CD1c+CD19- dendritic cells to CD1c+ dendritic cells were not significantly different between the three groups. CD40 (p=0.171), CD86 (p=0.037) and HLA-DR (p=0.036) were less expressed in the PACL than the BLD group. Expression of CD40 (p=0.319), CD86 (p=0.036) and HLA-DR (p=0.085) were less expressed in the PACL than the PSqCL group, but the differences were only significant for CD86. Expression of co-stimulatory markers was not different between the PSqCL and BLD groups. Expression of markers for activated DCs were dramatically lower in the PACL group than in groups with other histology (CD40 (p=0.005), CD86 (p=0.013) HLA-DR (p=0.004). Conclusion: These results suggest the possibility that CD1c+ myeloid DCs participate in control of the tumor immunity system and that low expression of markers results in lack of an immune response triggered by dendritic cells in adenocarcinoma of the lung.

Stability of $^{188}Re$ Labeled Antibody for Radioimmunotherapy and the Effect of Stabilizing Agents (방사면역치료용 $^{188}Re$ 표지 항체의 안정성과 안정제의 효과)

  • Chang, Young-Soo;Kim, Bo-Kwang;Jeong, Jae-Min;Chung, June-Key;Lee, Seung-Jin;Lee, Dong-Soo;Lee, Myung-Chul
    • The Korean Journal of Nuclear Medicine
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    • v.36 no.3
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    • pp.195-202
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    • 2002
  • Purpose: For clinical application of beta-emitter labeled antibody, high specific activity is imporiant. Carrier-free $^{188}Re$ from $^{188}W/^{188}Re$ generator is an ideal radionuclide for this purpose. However, low stability of $^{188}Re$ labeled antibody, especially in high specific activity, due to radiolytic decomposition by high energy (2.1 MeV) beta ray was problem. We studied the stability of $^{188}Re$ labeled antibody, and stabilizing effect of several stabilizers. Materials and Methods: Pre-reduced monoclonal antibody (CEA79.4) was labeled with $^{188}Re$ by incubating with generator-eluted $^{188}Re-perrhenate$ in the presence of stannous tartrate for 2 hr at room temperature. Radiochemical purity of each preparation was determined by chromatography. Human serum albumin was added to the labeled antibodies (2%). Stability of $^{188}Re-CEA79.4$ was investigated in the presence of ascorbic acid, ethanol, of Tween 80 as stabilizing agents. Results: Labeling efficiencies were $88{\pm}4%\;(n=12)$. Specific activities of $1.25{\sim}4.77MBq/{\mu}g$ were obtained. If stored after purging with $N_2$, all the preparations were stable for 10 hr. However, stability decreased in the presence of air. Perrhenate and $^{188}Re-tartrate$ was major impurity in declined preparation. colloid-formation was not a significant problem in all cases. Addition of ascorbic acid stabilized the labeled antibodies either under $N_2$ or under air by reducing the formation of perrhenate. Conclusion: High specific activity $^{188}Re$ labeled antibody is unstable, especially, in the presence of oxygen. Addition of ascorbic acid increased the stability.

Transformation of Populus nigra × P. maximowiczii Using Agrobacterium tumefaciens vectors (Agrobacterium tumefaciens vector를 이용(利用)한 양황철의 형질전환(形質轉換))

  • Son, Suk Gyu;Hyu, Jung Oh
    • Journal of Korean Society of Forest Science
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    • v.87 no.2
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    • pp.164-172
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    • 1998
  • This study was conducted to find the optimum transformation condition using Agrobacterium harboring promoterless GUS gene. The optimal medium for shoot induction from leaves of Populus nigra${\times}$P. maximowiczii was MS medium supplemented with $0.1mg/{\ell}$ NAA, $0.5mg/{\ell}$ BAP(94% regeneration frequency and 11.5 average number of shoot) According to the test using pBI121, the concentration of antibiotics for selection marker gene was $100mg/{\ell}$ kanamycin or $60mg/{\ell}$ geneticin in the SIM(shoot inducing medium) 3. Two weeks later, callus was induced in the SIM 3 and this callus grew up to 0.5-1cm shoots after 6 weeks in the new SIM 3. And the treatment with methylation inhibitor(5-azacytidine) led to a dramatic increase in foreign gene expression rate from 5.7% to 26.7%. The vector systems showed. different transformation efficiencies based on the fluorometric and histochemical GUS assay. In this study the vector systems used for transformation seemed to affect transformation frequency, in which pEHA101 yielded more transformants(35.9%) than LBA4404/pBI121 did(5.7%). This result indicated that pEHA101 was effective to insert the promoterless foreign gene into a poplar genome.

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Strategy for Bio-Diversity and Genetic Conservation of Forest Resources in Korea (생물종(生物種) 다양성(多樣性) 및 삼림유전자원(森林遺傳資源) 보존(保存) 전략(戰略))

  • Park, Young Goo
    • Journal of Korean Society of Forest Science
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    • v.83 no.2
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    • pp.191-204
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    • 1994
  • Due to its topographic complexities and various climatical condition, Korea exhibits diverse forest types. Dominant tree species in this zone are Quercus spp., Betula spp., Zelkova spp., Fraxinus spp., Pinus densiflora, Pinus koraiensis, and Pinus thunbergii ete. Genetic conservation in forest species in Korea there are three ways ; one is in situ, other is ex situ and third is in-facility conservation. In situ conservation include that are the present status of conservation of rare and endangered flora and ecosystem, the reserved forest, the national and provincial park, and the gene pool of natural forests. Ex situ conservation means to be established the new forest from in situ forest stands, progeny and provenance test populations, seed orchard and clone banks, and gene conservation in-facility. As a tool for low temperature storage, several aspects on in vitro system were studied ; (1) establishment of in vitro cultures from juvenile and/or rejuvenated tissues, (2) induction of multiple shoots from the individual micropropagules, (3) elongation of the proliferated shoots. Studies on cold storage for short-and long-term maintenance of in vitro cultures under $4^{\circ}C$ in the refrigerator were conducted. For the cryopreservation at $-196^{\circ}C$, various factors affecting survivability of the plant materials are being examined. The necessity of gene conservation of forest trees is enlarged not only to increase the adaptability for various environments but also to gain the breeding materials in the future. For effective gene conservation of forest trees, I would like to suggest followings ; 1. Forest stands reserved for other than the gene conservation purposes such as national parks should be investigated by botanical and gene-ecological studies for selecting bio-diversity and gene conservation stands. 2. Reserved forest for gene pool should be extented both economically important tree spp. and non-economical species. 3. Reserved forest for progeny test and clone bank should be systematically investigated for the use of Ex situ forest gene conservation. 4. We have to find out a new methodology of genetic analysis determining the proper and effective size of subpopulation for in situ gene conservation. 5. We should develop a new tree breeding systems for successful gene conservation and utilization of the genetic resources. 6. New method of in-facility gene conservation using advanced genetic engineering should be developed to save time and economic resources. 7. For the conservation of species with short-life span of seed or shortage of knowledge of seed physiology, tissue culture techniques will be played a great role for gene conservation of those species. 8. It is are very useful conservation not only of genes but of genotypes which were selected already by breeding program. 9. Institutional and administrative arrangements including legistlation must be necessarily taken for gene conservation of forest trees. 10. It is national problems for conservation of forest resources which have been rapidly destroyed because of degenerating environmental condition and of inexperienced management system of bio-diversity and gene conservation. 11. In order to international cooperation for exchanging data of bio-diversity and gene conservation, we should connect to international net works as soon as possible.

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A Study on the Applicability of Soilremediation Technology for Contaminated Sediment in Agro-livestock Reservoir (농축산저수지 오염퇴적토의 토양정화기술에 대한 적용성 연구)

  • Jung, Jaeyun;Chang, Yoonyoung
    • Journal of Environmental Impact Assessment
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    • v.29 no.3
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    • pp.157-181
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    • 2020
  • Sediments from rivers, lakes and marine ports serve as end points for pollutants discharged into the water, and at the same time serve as sources of pollutants that are continuously released into the water. Until now, the contaminated sediments have been landfilled or dumped at sea. Landfilling, however, was expensive and dumping at sea was completely banned due to the London Convention. Therefore, this study applied contaminated sedimentation soil of 'Royal Palace Livestock Complex' as soil purification method. Soil remediation methods were applied to pretreatment, composting, soil washing, electrokinetics, and thermal desorption by selecting overseas application cases and domestically applicable application technologies. As a result of surveying the site for pollutant characteristics, Disolved Oxigen (DO), Suspended Solid (SS), Chemical Oxygen Demand (COD), Total Nitrogen (TN), and Total Phosphorus (TP) exceeded the discharged water quality standard, and especially SS, COD, TN, and TP exceeded the standard several tens to several hundred times. Soil showed high concentrations of copper and zinc, which promote the growth of pig feed, and cadmium exceeded 1 standard of Soil Environment Conservation Act. In the pretreatment technology, hydrocyclone was used for particle size separation, and the fine soil was separated by more than 80%. Composting was performed on organic and Total Petroleum Hydrocarbon (TPH) contaminated soils. TPH was treated within the standard of concern, and E. coli was analyzed to be high in organic matter, and the fertilizer specification was satisfied by applying the optimum composting conditions at 70℃, but the organic matter content was lower than the fertilizer specification. As a result of continuous washing test, Cd has 5 levels of residual material in fine soil. Cu and Zn were mostly composed of ion exchange properties (stage 1), carbonates (stage 2), and iron / manganese oxides (stage 3), which facilitate easy separation of contamination. As a result of applying acid dissolution and multi-stage washing step by step, hydrochloric acid, 1.0M, 1: 3, 200rpm, 60min was analyzed as the optimal washing factor. Most of the contaminated sediments were found to satisfy the Soil Environmental Conservation Act's standards. Therefore, as a result of the applicability test of this study, soil with high heavy metal contamination was used as aggregate by applying soil cleaning after pre-treatment. It was possible to verify that it was efficient to use organic and oil-contaminated soil as compost Maturity after exterminating contaminants and E. coli by applying composting.

Detection of Osteomyelitis Using a Tc-99m Labeled Antigranulocyte Antibody Immunoscintigraphy (Tc-99m 표지 항과립구항체 면역신티그라피를 이용한 골수염의 진단)

  • Kang, Won-Jun;Chung, June-Key;Yeo, Jeong-Seok;Hong, Mee-Kyoung;Jeong, Jae-Min;Lee, Dong-Soo;Lee, Sang-Hoon;Choi, In-Ho;Lee, Myung-Chul
    • The Korean Journal of Nuclear Medicine
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    • v.32 no.4
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    • pp.344-353
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    • 1998
  • Purpose: The purpose of this study was to evaluate the diagnostic accuracy of Tc-99m labeled antigranulocyte antibody immunoscintigrapy in the diagnosis of osteomyelitis and compare with the results of triphasic bone scan. Materials and Methods: The study population was 39 patients (22 male, 17 female) who had uncertain diagnoses of osteomyelitis. Fifteen patients had history of orthopedic surgery, and 5 had previous fracture. One milligram of monoclonal antibody against NCA-95 was labeled with 370 MBq of Tc-99m, injected intravenously, and 4 hour images were obtained. Triphasic bone scan images were obtained in 30 patients. The final diagnosis was confirmed by bacteriologic culture, biopsy or long term clinical follow up. Results: Twenty one patients were confirmed to have osteomyelitis (1 acute, 20 chronic). Eighteen patients were without osteomyelitis. Antigranulocyte antibody immunoscintigraphy had a sensitivity of 71% (15/21), and a specificity of 89% (16/18), while the sensitivity and specificity of triphasic bone scan was 93% (13/14) and 38% (6/16), respectively. Antigranulocyte antibody scan showed higher specificity of 100% (11/11) in comparison with 33% (3/9) of triphasic bone scan in patients with history of orthopedic surgery or fracture. Conclusion: Antigranulocyte antibody immunoscintigraphy is more specific than that of triphasic bone scan and may be helpful in patients with history of surgery or fracture. However, sensitivity is lower than triphasic bone scan in the detection of chronic osteomyelitis.

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Evaluation of Heavy Metal Contamination in Streams within Samsanjeil and Sambong Cu Mining Area (삼산제일.삼봉 동광산 주변 수계의 중금속 오염도 평가)

  • Kim, Soon-Oh;Jung, Young-Il;Cho, Hyen-Goo
    • Journal of the Mineralogical Society of Korea
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    • v.19 no.3 s.49
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    • pp.171-187
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    • 2006
  • The status of heavy metal contamination was investigated using chemical analyses of stream waters and sediments obtained from Samsanjeil and Sambong Cu mining area in Goseong-gun, Gyeongsangnam-do. In addition, the degree and the environmental risk of heavy metal contamination in stream sediments was assessed through pollution index (Pl) and danger index (DI) based on total digestion by aqua regia and fractionation of heavy metal contaminants by sequential extraction, respectively. Not only the degree of heavy metal contamination was significantly higher in Samsanjeil area than in Sambong area, but its environmental risk was also revealed much more serious in Samsanjeil area than in Sambong area. The differences in status and level of contamination and environmental risk between both two mining areas may be attributed to existence of contamination source and geology. Acid mine drainage is continuously discharged and flows into the stream in Samsanjeil mining area, and it makes the heavy metal contamination in the stream more deteriorated than in Sambong mining area in which acid mine drainage is not produced. In addition, the geology of Samsanjeil mining area is mainly comprised of andesitic rocks including a small amount of calcite and having lower pH buffering capacity fer acid mine drainage, and it is likely that the heavy metal contamination cannot be naturally attenuated in streams. On the contrary, the main geology of Sambong mining area consists of pyroclastic sedimentary Goseong formation containing a high content of carbonates, particularly calcite, and it seems that these carbonates of high pH buffering capacity prevent the heavy metal contamination from proceeding downstream in stream within that area.

Evaluating the Impacts of Long-Term Use of Agricultural Chemicals on a Soil Ecosystem by Structural Analysis of Bacterial Community (세균군집의 구조분석을 통한 장기간 농약사용이 토양생태계에 미치는 영향 평가)

  • Yun, Byeong-Jun;Kim, Seong-Hyeon;Lee, Dong-Heon;O, Gye-Heon;Gang, Hyeong-Il
    • Korean Journal of Microbiology
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    • v.39 no.4
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    • pp.260-266
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    • 2003
  • In this study bacterial community was analyzed to evaluate the impacts of long-term use of agricultural chemicals on a soil ecosystem as well as to obtain fundamental data on the relationship. Sequences of 16S rRNA clones from a non-agricultural site and a tangerine orchard soil which has a history of long-term use of agricultural chemicals over 30 years were analyzed. This revealed that bacterial community containing 5 divisions and 18 genera was distributed in a tangerine orchard soil, while bacterial community containing 9 divisions and 44 genera was distributed. In a tangerine orchard soil site, the most abundant bacteria in subdivision level were placed into Proteobacteria γ group which occupied 56% of total clones. The other bacterial clones from the ocrhcard soil exposed to agricultural chemicals over 30 years were Acidobacteria group (25%), Fimicutes group (5%), Planctomycetes group (2%), Proteobacteria α (1%), δ group (1%), and Cyanobacteria group (1%). Whereas, the clones were from the non-agricultural site were distributed among the division or subdivision Acidobacteria group (14%), Planctomycetes group (13%), Proteobacteria α (10%), β (9%), δ (9%), Fimicutes group (8%), Verrucomicrobia group (8%), Actinobacteria group (6%), Proteobacteria γ group (3%), Bacteroidetes group (3%), Gemmatimonadetes group (3%), and Cyanobacteria group (1%). This finding suggests the possibility that long-term application of agricultural chemicals or fertilizers on a tangerine orchard might result in drastic reduction or alteration in the composition of the bacterial community in the contaminated soil site.

Characterization of Ferritin Isolated from Dog Spleen (개의 비장에서 분리한 페리틴의 특성)

  • Park Jae-Hag;Jun Do Youn;Kim Young Ho
    • Journal of Life Science
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    • v.15 no.3 s.70
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    • pp.439-446
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    • 2005
  • Ferritin is known to be the principle iron-storage protein in a wide variety of rganisms. The electro­phoretic mobility and immunological cross-reactivity of dog splenic ferritin were compared with those of horse, bovine, and pig splenic ferritin after isolation using heat treatment, salting out, column chromatography, and ultrafiltration. These isolation methods allowed the recovery of $\~84{\mu}g$ of the ferritin per g of spleen. The iron content in the dog ferritin was $22.7\%$, which appeared to be higher than those in the other mammalian ferritins tested. The electrophoretic mobility of the dog ferritin under nondenaturing conditions was similar to its bovine counterpart, whereas it was more identical to pig and horse ferritins on an SDS-polyacrylamide gel. The molecular weight of the dog ferritin subunit was 19.5 kDa on an SDS-polyacrylarnide gel, and the subunit was unable to bind with iron. The polyclonal anti-dog ferritin raised in rats was able to cross-react with the pig, bovine, and horse ferritins, upon Ouchterlony double immunodiffusiion. A Western blot analysis also revealed that the anti-dog ferritin, which specifically bound with the dog ferritin subunit, could also recognize the horse, bovine, and pig ferritin subunits and the maximum cross-reactivity was exhibited with the pig ferritin subunit, indicating that the dog ferritin is immunochemically more similar to the pig ferritin than its other mammalian counterparts. Accordingly, these results elucidate the biochemical and immunochemical characteristics of dog ferritin that might have a potential to be applied as an oral iron supplement to treat iron deficiency anemia.

Genes of Wild Rice (Oryza grandiglumis) Induced by Wounding and Yeast Extract (상처와 효모추출물 처리조건에서 유발되는 야생벼 유전자 스크린)

  • Shin, Sang-Hyun;Im, Hyun-Hee;Lee, Jai-Heon;Kim, Doh-Hoon;Chung, Won-Bok;Kang, Kyung-Ho;Cho, Sung-Ki;Shin, Jeong-Sheop;Chung, Young-Soo
    • Journal of Life Science
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    • v.14 no.4
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    • pp.650-656
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    • 2004
  • Oryza grandiglumis (CCDD, 2n=48), one of the wild rice species, has been known to possess fungal-,bacterial-, and insect-resistance against sheath blight, rice blast, bacterial leaf blight and brown plant hopper (Nilaparvata lugens). To rapidly isolate differentially expressed genes responding to fungal and wounding stress, wounding and yeast extract were treated to O. grandiglumis for 24 hrs. Suppression subtractive hybridization (SSH) method was used to obtain differentially expressed genes from yeast extract and wounding treated plants. Seven hundreds and seventy six clones were obtained by subcloning PCR product, and colony array and screening were carried out using radio-isotope labeled cDNA probes prepared from the wounding and yeast extract treated plants. One hundred and fifteen colonies were confirmed as true positive ones. Average insert size of the clones were ranged from 400 bp to 700 bp and all the inserts were sequenced. To decide the identity of those clones, sequences were analyzed by sequence homology via GenBank database. The homology search result showed that 68 clones were matched to the genes with known function; 16 were related to primary metabolism, 5 to plant retrotransposons, 5 to defense related metallothionein-like genes. In addition to that, others were matched to various genes with known function in amino acid synthesis and processing, membrane transport, and signal transduction, so on. In northern blot analysis, induced expressions of ogwfi-161, ogwfi-646, ogwfi-663, and ogwfi-695 by wounding and yeast extract treatments were confirmed. The result indicates that SSH method is very efficient for rapid screening of differentially expressed genes.