• Title/Summary/Keyword: GC-1

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Expression of IgG1 Germline Transcripts in Germinal Center B Cells of Mouse Popliteal Lymph Nodes

  • Lee, In-Woo;Kim, Jin-Ho;Chung, Gook-Hyun
    • BMB Reports
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    • v.29 no.2
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    • pp.127-132
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    • 1996
  • Germinal centers (GCs) are formed in peripheral lymphoid tissues in response to protein antigens. In order to see if immunoglobulin isotype switching takes place in GC B-cells, we isolated GC B-cells (PNA positive cells) from mouse popliteal lymph nodes by a flow cytometer after the staining of lymph node cells with PNA-FITC and anti-B220-PE, and determined the expression of ${\gamma}1$ germline transcript and ${\gamma}1$ mRNA by RT-PCR. ${\gamma}1$ germline transcript and ${\gamma}1$ mRNA were amplified specifically in cDNAs from hybridoma expressing IgG1 or splenocytes stimulated LPS plus IL-4. Germinal center B-cells formed in popliteal lymph nodes of mice immunized with chicken ovalbumin were isolated 7 days after immunization. We sorted GC B-cells five times. Immunoglobulin ${\gamma}1$ germline transcripts were expressed in germinal center B-cells in three out of five sorts whereas two out of five sorts did not express ${\gamma}1$ germline transcripts in GC B-cells. The contents of GC B-cells ranged from 5 to 7% of total lymph node cells in most flow cytometric analyses but those of two sorted cells which did not express ${\gamma}1$ germline transcripts were out of normal range. These results imply that isotype switching of immunoglobulins may take place in GCs.

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Methylation Levels of LINE-1 As a Useful Marker for Venous Invasion in Both FFPE and Frozen Tumor Tissues of Gastric Cancer

  • Min, Jimin;Choi, Boram;Han, Tae-Su;Lee, Hyuk-Joon;Kong, Seong-Ho;Suh, Yun-Suhk;Kim, Tae-Han;Choe, Hwi-Nyeong;Kim, Woo Ho;Hur, Keun;Yang, Han-Kwang
    • Molecules and Cells
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    • v.40 no.5
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    • pp.346-354
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    • 2017
  • Long interspersed nuclear element-1 (LINE-1) is a retrotransposon that contains a CpG island in its 5'-untranslated region. The CpG island of LINE-1 is often heavily methylated in normal somatic cells, which is associated with poor prognosis in various cancers. DNA methylation can differ between formalin-fixed paraffin-embedded (FFPE) and frozen tissues. Therefore, this study aimed to compare the LINE-1 methylation status between the two tissue-storage conditions in gastric cancer (GC) clinical samples and to evaluate whether LINE-1 can be used as an independent prognostic marker for each tissue-storage type. We analyzed four CpG sites of LINE-1 and examined the methylation levels at these sites in 25 FFPE and 41 frozen GC tissues by quantitative bisulfite pyrosequencing. The LINE-1 methylation status was significantly different between the FFPE and frozen GC tissues (p < 0.001). We further analyzed the clinicopathological features in the two groups separately. In the frozen GC tissues, LINE-1 was significantly hypomethylated in GC tissues compared to their corresponding normal gastric mucosa tissues (p < 0.001), and its methylation status was associated with gender, differentiation state, and lymphatic and venous invasion of GC. In the FFPE GC tissues, the methylation levels of LINE-1 differed according to tumor location and venous invasion of GC. In conclusion, LINE-1 can be used as a useful methylation marker for venous invasion in both FFPE and frozen tumor tissues of GC.

CircCOL1A2 Sponges MiR-1286 to Promote Cell Invasion and Migration of Gastric Cancer by Elevating Expression of USP10 to Downregulate RFC2 Ubiquitination Level

  • Li, Hang;Chai, Lixin;Ding, Zujun;He, Huabo
    • Journal of Microbiology and Biotechnology
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    • v.32 no.7
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    • pp.938-948
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    • 2022
  • Gastric cancers (GC) are generally malignant tumors, occurring with high incidence and threatening public health around the world. Circular RNAs (circRNAs) play crucial roles in modulating various cancers, including GC. However, the functions of circRNAs and their regulatory mechanism in colorectal cancer (CRC) remain largely unknown. This study focuses on both the role of circCOL1A2 in CRC progression as well as its downstream molecular mechanism. Quantitative polymerase chain reaction (qPCR) and western blot were adopted for gene expression analysis. Functional experiments were performed to study the biological functions. Fluorescence in situ hybridization (FISH) and subcellular fraction assays were employed to detect the subcellular distribution. Luciferase reporter, RNA-binding protein immunoprecipitation (RIP), co-immunoprecipitation (Co-IP), RNA pull-down, and immunofluorescence (IF) and immunoprecipitation (IP) assays were used to explore the underlying mechanisms. Our results found circCOL1A2 to be not only upregulated in GC cells, but that it also propels the migration and invasion of GC cells. CircCOL1A2 functions as a competing endogenous RNA (ceRNA) by sequestering microRNA-1286 (miR-1286) to modulate ubiquitin-specific peptidase 10 (USP10), which in turn spurs the migration and invasion of GC cells by regulating RFC2. In sum, CircCOL1A2 sponges miR-1286 to promote cell invasion and migration of GC by elevating the expression of USP10 to downregulate the level of RFC2 ubiquitination. Our study offers a potential novel target for the early diagnosis and treatment of GC.

The Effects of Sb Additions on the Structures and Mechanical Properties of C/V Graphite Cast Iron (C/V 흑연주철(黑鉛鑄鐵)의 조직(組織)과 기계적성질(機械的性質)에 미치는 Sb첨가효과(添加效果))

  • Choi, Yang-Jin;Kang, In-Chan
    • Journal of Korea Foundry Society
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    • v.7 no.1
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    • pp.53-59
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    • 1987
  • In order to study the infuence of Sb addition on the compacted vermicular graphite cast iron(C/V) and gray cast iron(GC), these specimen which had different Sb content(0-0, 1wt.%) were prepared. these specimen were tested for microstructure and mechanical properties. The result from this study are as follows. For C/V and GC, the microstructure of graphite appeared to be finer with increasing Sb content more than 0.06wt.% Ferrite disappeared from the matrix with addition of Sb more than 0.06wt.% in GC and more than 0.08wt.% in C/V. The size of the eutectic cell in GC was 4.5 to 7 times larger than that in C/V, but the decreasing tendancy of its size was more remarkable in GC than in C/V with increasing Sb content. The abrasion speed corresponding to the maximum amount of abrasion in the nabrasion test was found to be 1.37m/sec. GC sample weared 3.6 to 21 times faster than those of C/V did. the amount of abrasion demeased remarkably with Sb addition, and the abrasion rate was constant with Sb content over 0.04wt.%

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Analysis of the Taste Components and Antioxidant Properties of Cheonggukjang Containing Korean Red Ginseng

  • Kim, Eun-Jung;Hong, Ju-Yeon;Shin, Seung-Ryeul;Moon, Yong-Sun;Yoon, Kyung-Young
    • Food Science and Biotechnology
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    • v.18 no.1
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    • pp.53-59
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    • 2009
  • This study was performed to investigate the taste composition and antioxidant properties of cheonggukjang containing Korean red ginseng (RGC), as compared to either general cheonggukjang (GC) or non-fermented boiled soybeans (BS). Amylase activity was the highest (576.7 unit/g) in RGC, whereas protease activity was the highest (326.0 unit/g) in GC. The total soluble sugar contents of BS, GC, and RGC were 2,027.5, 905.5, and 837.5 mg/100 g, respectively. RGC had the highest amount of total amino acids (2,127.4 mg/100 g) and essential amino acid (50.9%) among the samples. The ratio of sweet to bitter components was higher in RGC than in GC. Although the extracts of RGC had higher radical scavenging activity for 1,1-diphenyl-2-picrylhydrazyl (DPPH) than BS or GC, regardless of the extract concentration, the ethanol extract of RGC showed the highest scavenging ability (92.4%) at 2.0 mg/mL. The chloroform extracts from GC and RGC showed their greatest superoxide dimutase-like activities at 17.2 and 19.7% at a concentration of 2 mg/mL, respectively. Regardless of the samples, the nitrite scavenging ability was positively correlated to the extract concentration, and RGC had highest ability among samples under the same extract concentrations.

Studies on disease of catfish(Silurus asotus) in Korea Pathology of skin ulcerous by A. hydrophila (한국산메기(Silurus asotus)의 질병에 관한 연구)

  • 한규삼;최인열;배정준;김영길;이근광;김영진
    • Korean Journal of Veterinary Service
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    • v.16 no.2
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    • pp.103-110
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    • 1993
  • The Aeromonas strains were isolated from diseased cultured catfsh (Silurus asotus) in the ponds near by Gochang of Chunbuk. The present isolates were identified as A. hydrophila based on their biological and biochemica1 characteristics. The isolates of A. hydrophila were named GC-1, GC-2, GC-3, GC-4 and GC-5, Five strains were grew optimally at temperatures $35^{\circ}C,$ pH 7.5 and in 0.5 to l% NaCl. This bacterium(GC-1) was injected into health catfish in order to prove the causative agent of ascites and haemorrhagic ulcers. The symptoms in cat fish infected by this challenge method were observed to be very similar to the symptoms of a natural infection, but controls did not show any abnormal symptoms during the experimental period. At 24 post-injection, the red spot developed around the injection site and the haemorrhagic ulcers was extended near the gill and ventral fin. Five strains were tested for drug sensitivity by plate method. All strains were sensitivity to gentamicin and resistant to penicillin, streptomycin and tetracycline.

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Analytical Characteristics of GC/MS and HPLC according to the Concentration Distribution of PAHs (PAHs 농도 분포에 따른 GC/MS와 HPLC의 분석특성에 관한 연구)

  • Hong, Jwa-Ryung;Choi, Kwang-Min
    • Journal of Korean Society of Occupational and Environmental Hygiene
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    • v.25 no.3
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    • pp.312-321
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    • 2015
  • Objectives: The purpose of this study was to determine the best method to analyze PAHs at extremely low concentrations. To this end, 16 PAHswere analyzed simultaneously by GC/MS, HPLC/FLD and HPLC/UVD, and the analytical characteristics of HPLC and GC/MS were compared. Methods: This study was conducted by GC/MS and HPLC/FLD/UVD, and evaluated linearity, precision and detection limit. Standard solutions were prepared for 21 samples in the range of $0.00001{\sim}1.0{\mu}g/mL$ and the samples were divided into four groups. All samples were made in three sets and analysis was replicated seven times. Results: Sixteen PAHs could be simultaneously separated by HPLC and GC/MS, and the adequate equipment was HPLC/FLD. The retention times by HPLC were shorter than GC/MS, and HPLC had better separation for most PAHs than GC/MS. The peaks of naphthalene and naphthalene-D8 partially overlapped for GC/MS. HPLC/FLD had a 20-2000 times lower limit of detection than GC/MS and UVD. However FLD was not adequate for analyzing acenaphthylene because it has too low a fluorescence quantum yield to be detected. The precision of HPLC/FLD/UVD and GC/MS showed less than 20% at $0.001{\mu}g/mL$ PAHs and when the concentration was higher, the coefficient of variation was decreased. HPLC/FLD was better for the overall detection of limits. Conclusions: The results indicate that the HPLC/FLD method has good linear range, precision and a detection of limits from $0.00001{\sim}0.0001{\mu}g/mL$ for all 16 PAHs. This study contributes to providing useful data for analysis technology and can be applied to occupational exposure measurement for PAHs in workplaces.

Simultaneous Analysis of Ethanol and Harmful Components by GC in Alcoholic Liquors (GC를 이용한 주류중 에탄올 및 유해 성분의 동시 분석법)

  • 정의한;이명자;강금자;문혜성;유병옥;황정구;장준식
    • Journal of Food Hygiene and Safety
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    • v.7 no.1
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    • pp.45-48
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    • 1992
  • In Food revolution of Korea, colorimetries or a titration methods are introduced for the analysis of alcoholic liquors. But, these wet analyses have disadvantages such as individual errors, long process time, and sometimes tedious pretreatments. In the process of making alcoholic liquors, fusel oils are produced as by products. Five main fusel components that could be produced are 2-propanol, n-propanol, iso-butanol, n-butanol, and isoamyl alcohol. Also acetaldehyde and methanol could be produced as by-products of ethanol. With using capillary FFAP column in GC or GC/MSD, we analysed these five fusel components as well as internal standard (acetonitrile) including methanol, acetaldehyde and ethanol simultaneously. We obtained excellent mass spectra as qualitative data of all species. We also took excellent quantitative data with GC by using the internal standard method.

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Comparative Sampling Procedures for the Volatile Flavor Components of Codonopsis lanceolata (전처리 방법에 따른 더덕(Codonopsis lanceolata)의 휘발성 향기성분 비교 분석)

  • Kim, Jung-Han;Kim, Kyoung-Rae;Kim, Jae-Jung;Oh, Chang-Hwan
    • Korean Journal of Food Science and Technology
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    • v.24 no.2
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    • pp.171-176
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    • 1992
  • Volatile flavor components of Codonopsis lanceolata were extracted by gas co-distillation (GCD), solvent extraction/fractionation (SEF), and headspace sampling (HSS) methods. The extracts were analyzed by dual-capillary gas chromatography-retention index (GC-RI) and gas chromatography-mass spectrometry(GC-MS). The two extracts prepared by SEF and HSS gave more similar fragrance to the Codonopsis lanceolata than the GCD extract. The GC profiles of the SEF and HSS extracts were similar to each other except for differences in peak areas. The extract prepared by SEF gave a sweet note while the extract prepared by HSS gave a green note. The GCD extract began to give a burnt note of herb medicine with prolonged distillation. Rapid extraction of flavor components from Codonopsis lanceolata was possible in several short steps by SEF and HSS methods compared to GCD. GC-MS and GC-RI were used for peak identification. GC-RI was more effective for identification of isomers, and polar FFAP column was more suitable for identification of polar compounds. From Codonopsis lanceolata we identified 35 volatile flavor constituents, 24 of which have not been previously reported by simultaneous distillation extraction method $^{(5)}$. trans-2-Hexanal, cis-3-hexen-1-ol, trans-2-hexen-1-ol, and hexanol were considered key components of the green note and 1-octen-3-ol, the component of the fresh note. Esters, including amyl propionate, seem to be responsible for the sweet note particular to Codonopsis lanceolata.

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Evaluation of Biogas Production Rate by using Various Electrodes Materials in a Combined Anaerobic Digester and Microbial Electrochemical Technology (MET) (미생물 전기화학 기술이 적용된 단일 혐기성소화조에서 전극재질에 따른 바이오가스 생성 효율 평가)

  • Shin, Wonbeom;Park, Jungyu;Lee, Beom;Kim, Yonggeun;Jun, Hangbae
    • Journal of Korean Society of Environmental Engineers
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    • v.39 no.2
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    • pp.82-88
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    • 2017
  • MET (Microbial Electrochemical Technology), such as MFC (Microbial Fuel Cell) and MEC (Microbial Electrolysis Cell), is a promising technology for producing sustainable biogas from an anaerobic digester (AD). At current stage, however, the most likely limiting factors, large internal resistances, should be overcome for successful scale up of this technology. Various researchers reported that application of electrode materials containing high current density, increase of ion strength and conductivity, configuration of electrode are good methods for minimizing internal resistances. Recently, stainless steel is receiving great attention because of not only high performance and durability but also low cost. Therefore, in this study, we evaluate electrochemical characteristics and biogas production rate using various electrode materials and configuration (graphite carbon coated with catalysts ($GC-C_M$) or not (GC), stainless steel mesh (SUS-M) and plate (SUS-P)). As the results, current densities of $GC-C_M$, GC, SUS-P, SUS-M were 2.03, 1.36, 1.04, $1.13A/m^2$, respectively. Methane yields of $GC-C_M$, GC, SUS-P, SUS-M were 0.27, 0.14, 0.19, 0.21 $L-CH_4/g-COD_{rem}$., respectively. Stainless steel shows high current density and methane yield, which are similar as graphite carbon coated with catalysts.