• Title/Summary/Keyword: semi-quantitative analysis

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Identification of Interleukin 1-Responsive Genes in Human Chondrosarcoma SW1354 cells by cDNA Microarray Technology

  • Jeon, Jun-Ha;Jung, Yong-Wook;Yun, Dae-Young;Kim, Hyun-Do;Kwon, Chang-Mo;Hong, Young-Hoon;Kim, Jae-Ryong;Lee, Choong-Ki
    • Journal of Yeungnam Medical Science
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    • v.24 no.1
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    • pp.24-40
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    • 2007
  • Background : Accumulating evidence shows that interleukin(IL)-1 plays a critical role in inflammation and connective tissue destruction observed in both osteoarthritis and rheumatoid arthritis. IL-1 induces gene expression related to cytokines, chemokines and matrix metalloproteinases by activation of many different transcription factors. Materials and Methods : The chondrosarcoma cell line, SW1353, is known to be a valuable in vitro system for investigating catabolic gene regulation by IL-$1{\beta}$ in chondrocytic cells. To explore and analyze the changes in gene expression by IL-1 responsible for arthritis, SW1353 was treated with IL-1 for 1, 6 and 24 h and then total RNAs were purified for each time. The changes in gene expression were analyzed with 17k human cDNA microarrays and validated by semi-quantitative RT-PCR. Results : Greater than a two-fold change was observed in 1,200 genes including metallothioneins, matrix metalloproteinases, extracellular matrix proteins, antioxidant proteins, cytoskeleton proteins, cell cycle regulatory proteins, proteins for cell growth and apoptosis, signaling proteins and transcription factors. These changes appeared to be correlate with the pathophysiological changes observed in early osteoarthritis. Conclusion : cDNA microarray analysis revealed a marked variability in gene expression, and provided insight into the overall molecular changes. The result of this study provide initial information for further studies to identify therapeutic targets in osteoarthritis pathogenesis.

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Usefulness of Low Dose Oral Contrast Media in $^{18}F-FDG$ PET/CT ($^{18}F-FDG$ PET/CT에서 저용량 경구용 조영제의 유용성)

  • An Young-Sil;Yoon Joon-Kee;Hong Seon Pyo;Joh Chul-Woo;Yoon Seok-Nam
    • Nuclear Medicine and Molecular Imaging
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    • v.40 no.5
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    • pp.257-262
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    • 2006
  • Purpose: The standard protocol using large volume of oral contrast media may cause gastrointestinal discomfort and contrast-related artifacts in PET/CT. The aim of this study was to evaluate the usefulness of low dose oral contrast in $^{18}F-FDG$ PET/CT. Materials and Methods: We retrospectively reviewed the whole-body PET/CT images in a total of 435 patients. About 200 ml of oval contrast agent (barium sulfate) was administered immediately before injection of $^{18}F-FDG$. The FDG uptake of intestines was analyzed by visual and semi- quantitative method on transaxial, coronal and saggital planes. Results: Seventy (16%, 113 sites) of 435 images showed high FDG uptake (peak SUV > 4); 50 (74%, 84 sites) with diffuse and 20 (15%, 29 sites) with focal uptake. The most commonly delivered site of oral contrast media was small bowel (n=27, 39%). On PET/CT images, FDG uptake coexisted with oral contrast media in 26 patients (54%, 38 sites) with diffuse pattern and 9 (45%, 9 sites) with focal pattern, and by sites, those were 38 (45%) and 9 (31%), respectively. In small bowel regions, the proportion of coexistence reached as high as 61% (29/47 sites). A visual analysis of available non-attenuation corrected PET images of 27 matched regions revealed no contrast-related artifact. Conclusion: We concluded that the application of low dose contrast media could be helpful in the evaluation of abdominal uptake in the FDG PET/CT image.

Novel quantitative trait loci for the strong-culm and high-yield related traits in rice detected from the F2 population between the super thick-culm and super grain-bearing line 'LTAT-29' and the high-yielding variety 'Takanari'

  • Nomura, Tomohiro;Yamamoto, Toshio;Ueda, Tadamasa;Yonemaru, Junichi;Abe, Akira;Adachi, Shunsuke;Hirasawa, Tadashi;Ookawa, Taiichiro
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.95-95
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    • 2017
  • Lodging is a serious issue in rice production, because it drastically decreases the biomass production and grain yield. Since the Green Revolution, the lodging resistance has been increased by lowering the moment of above-ground parts due to the short culm by the semi-dwarf gene sd1. However, it has been pointed out that sd1 alone has suppressive effects for biomass production and yield. To increase rice yield, the long-culm and large panicle type varieties with a superior lodging resistance need to be developed. To improve the lodging resistance and yield of these type varieties, it would be effective to identify novel alleles for these traits underlying natural variations in rice and to pyramid these alleles to a single rice variety. In order to perform this strategy, we have developed new rice lines derived from crosses among varieties with superior alleles. At first, TULT-gh-5-5 was selected from a cross between strong culm and high biomass variety Leaf Star and high-yielding variety Takanari, and TUAT-32HB was selected from a cross between high-yielding variety Akenohoshi and Takanari. Then, we developed the super thick-culm and super grain-bearing line, LTAT-29 derived from a cross between TULT-gh-5-5 and TUAT-32HB. In the current study, to identify the QTLs and genes relating to the strong culm and the high yield of LTAT-29, we performed QTL analysis using SNPs markers with $F_2$ population derived from a cross between LTAT-29 and Takanari. LTAT-29 has never lodged throughout the growth period despite it had long culms and heavy panicles. LTAT-29 had a larger outer diameter of the culm and twice the size of the section modulus than Takanari. As a result, the bending moment at breaking of LTAT-29 was significantly larger than that of Takanari. Brown rice yield of LTAT-29 was $9.2t\;ha^{-1}$ about 10% higher than that of Takanari due to the larger number of spikelets per panicle. LTAT-29 had a greater number of secondary branches per panicle. In the $F_2$ population between LTAT-29 and Takanari, we found continuous frequency distributions in the section modulus and the spikelet number per panicle. Two QTLs increased the section modulus by the alleles of LTAT-29 were detected on Chr.1L and Chr.2L. One QTL increased the spikelet number per panicle of Takanari by the allele of LTAT-29 was detected on Chr.1L, and two QTLs increased the number of secondary branches per panicle by the alleles of LTAT-29 were detected on Chr.1L and Chr.4L. It was found that the alleles of these QTLs were the japonica type originated from Leaf Star or Akenohoshi. The novel QTLs for the traits related to super thick-culm and super grain-bearing and their combinations could be utilized for improving the lodging resistance and yield in rice varieties.

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$^{99m}Tc$-Glucarate Uptake in Ischemic Tissue of Experimental Models of Cerebral Ischemia (실험적 뇌허혈증 모델에서 허혈 조직의 $^{99m}Tc$-glucarate 섭취)

  • Jeong, Jae-Min;Kim, Young-Ju;Choi, Seok-Rye;Kim, Chae-Kyun;Mar, Woong-Chun;Chung, June-Key;Lee, Myung-Chul;Koh, Chang-Soon;Lee, Dong-Soo
    • The Korean Journal of Nuclear Medicine
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    • v.30 no.4
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    • pp.484-492
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    • 1996
  • To detect ischemic tissue in experimental model of cerebral ischemia made by middle cerebral artery(MCA)-occlusion, we acquired triple image of $^{99m}Tc$-glucarate, [$^{18}F$]fluoro-deoxyglucose (FDG), and 2,3,5- triphenyltetrazolium (TTC) staining. We made cerebral infarction either with reperfusion (after occlusion of 2 hours) or without reperfusion in 10 Sprague-Dawley rats by inserting thread to MCA through internal carotid artery. After 22 hours, we injected 740 MBq of $^{99m}Tc$-glucarate and 55.5 MBq of [$^{18}F$]FDG through tail vein. Each 1 mm slice of rat brains was frozen and exposed to imaging plate for 20 minutes in freezer to get an [$^{18}F$]FDG image. After 20 hours enough to fade radioactivity of [$^{18}F$]FDG, the slices were again imaged by BAS1500 for $^{99m}Tc$-glucarate uptake. Finally, these brain tissues were stained with TTC. Semi-quantitative visual analysis was done by grading 0 to 3 points according to the degree of uptakes($^{99m}Tc$-glucarate) and decreased uptakes([$^{18}F$]FDG and TTC). Ten rats survived with neurologic symptoms. TTC staining confirmed the development of infarction. The size of the infarction was relatively larger in the group without reperfusion. [$^{18}F$]FDG images were similar to TTC-stained images. However, we found regions with intermediate uptake which were not stained with TTC. We found regions with intermediate [$^{18}F$]FDG uptake where TTC staining was normal. $^{99m}Tc$-glucarate uptake was round only in TTC non-stained region. In the TTC stained regions, there were no uptake of $^{99m}Tc$-glucarate. We could not find clear relation between $^{99m}Tc$-glucarate uptake with [$^{18}F$]FDG uptake. This was partly because percent uptake of $^{99m}Tc$-glucarate was so small (less than 1 percent of injected dose) and because there were quite heterogeneity of patterns of [$^{18}F$]FDG uptake and TTC. With these findings, we could conclude that $^{99m}Tc$-glucarate were taken up only in part of ischemic tissues which were proven to be nonviable. The establishment of MCA-occluded rat model with or without reperfusion and triple imaging for $^{99m}Tc,\;^{18}F$ and TTC helped the characterization of $^{99m}Tc$-glucarate uptakes. Further work is needed to clarify the meaning or diversities or [$^{18}F$]FDG and TTC and their relation with $^{99m}Tc$-glucarate.

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Validation and Calibration of Semi-Quantitative Food Frequency Questionnaire - With Participants of the Korean Health and Genome Study - (반정량식품섭취빈도조사지의 타당성 검증 및 보정 - 지역사회 유전체 코호트 참여자를 대상으로 -)

  • Ahn, Youn-Jhin;Lee, Ji-Eun;Cho, Nam-Han;Shin, Chol;Park, Chan;Oh, Berm-Seok;Kimm, Ku-Chan
    • Korean Journal of Community Nutrition
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    • v.9 no.2
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    • pp.173-182
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    • 2004
  • We carried out a validation-calibration study of the food frequency questionnaire (FFQ) that we had previously developed for a community-based cohort of the Korean Genome and Health Study of the Korea National Genome Research Institute. We have collected a total of 254 3-day diet records (DRs) from 400 subjects, 200 each randomly selected from the two study cohorts of Ansung and Ansan. FFQ was administered at the time of cohort recruitment in 2001, and DRs were collected during a two month period from January through February of 2002. The mean age was 52.2 years. Farming for men and housewife for women were the most common occupations. The majority of the subjects had undergone 6∼12 years of education. The general characteristics including demographic and other data were not different from the total cohort subjects. Absolute levels of consumed nutrients including total energy (energy), protein, fat, carbohydrate, calcium, phosphorus, sodium, potassium, iron, retinol, carotene, vitamin A, thiamin, riboflavin, niacin and vitamin C were compared. The average of energy intake was not significantly different between the data collected by the 2 methods. However, consumptions of protein and fat were higher in data of DRs, whereas that of carbohydrate was higher in FFQ data. Significant correlation of each nutrient consumption between the data sets was observed (p < 0.05) except in the case of iron, while the average correlation coefficient between them was 0.22 ranging from 0.33 for energy to 0.11 for iron. The results of cross classification by quantile for exact classification ranged from 25.2% (carotene) to 35.0% (phosphorus), and from 64.6% (vitamin A) to 76.4% (retinol) for adjacent classification. The proportion of completely opposite classification was 8.1% in average. Calibration slope was estimated by regression and calibration parameters ranged from 0.025 for carotene to 0.423 for niacin. We conclude that the FFQ we have developed is an appropriate tool for assessing the nutrient intakes as ranking exposures in epidemiology studies in view that amounts of consumed nutrients obtained by FFQ were similar to those collected by DRs, that correlations between consumed nutrients collected by these methods were significant, and that classification results were relatively fair. The correlation coefficients, however, were lower than expected, which may be mainly due to the survey season. In fact, any short-term dietary survey cannot accurately reflect the overall dietary intakes that change heavily depending on seasons. Further studies including the analysis of chemical indices would be helpful for the studies of causal relationship between the diet and disease.

Expression of Cell Cycle Related Genes in HL60 Cells Undergoing Apoptosis by X-irradiation (HL60 세포주에서 방사선 조사에 의한 Apoptosis와 세포 주기 관련 유전자의 발현 변화)

  • Kim, Jin-Hee;Park, In-Kyu
    • Radiation Oncology Journal
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    • v.16 no.4
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    • pp.377-388
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    • 1998
  • Purpose : To evaluate changes in expression of cell cycle related genes during apoptosis induced in HL60 cells by X-irradiation to understand molecular biologic aspects in mechanism of radiation therapy. Material and Methods : HL-60 cell line (promyelocytic leukemia cell line) was grown in culture media and irradiated with 8 Gr by linear accelerator (6 MV X-ray). At various times after irradiation, ranging from 3 to 48 hours were analyzed apoptotic DNA fragmentation assay for apoptosis and by western blot analysis and semi-quantitative RT-PCR for expression of cell cycle related genes (cyclin A, cyclin B, cyclin C, cyclin Dl, cyclin E, cdc2, CDK2, CDK4, $p16^{INK4a}$, $p21^{WAF1}$, $p27^{KIP1}$, E2F, PCNA and Rb). Results : X-irradiation (8 Gy) induced apoptosis in HL-60 cell line. Cycline A protein increased after reaching its peak 48 h after radiation delivery and cyclin E, E2F, CDK2 and RB protein increased then decreased after radiation. Radiation induced up-regulation of the expression of E2F is due to mostly increase of Phosphorylated retinoblastoma proteins (ppRb). Cyclin Dl, PCNA, CDC2, CDK4 and $p16^{INK4a}$ protein underwent no significant change at any times after irradiation. There was not detected $p21^{WAF1}$ and $p27^{KIP1}$ protein. Cyclin A, B, C mRNA decreased immediately after radiation and then increased at 12 h after radiation. Cyclin Dl mRNA increased immediately and then decreased at 48 h after radiation. After radiation, cyclin E mRNA decreased with the lapse of time. CDK2 mRNA decreased at 3h and increased at eh after radiation. CDK4 mRNA rapidly increased at 6 to 12 h after radiation. There was no change of expression of $p16^{INK4a}$ and not detected in expressin of $p21^{WAF1}$ and $p27^{KIP1}$ mRNA. Conclusion : We suggest that entry into S phase may contribute to apoptosis of HL60 cells induced by irradiation. Increase of ppRb and decrease of pRb protein are related with radiation induced auoptosis of HL60 cells and tosis of HL60 cells induced by irradiation. Increase of ppRb and decrease of PRb protein are related with radiation induced apoptosis of HL60 cells and this may be associated with induction of E2F and cyclinE/CDK2. These results support that $p21^{WAF1}$ and $p27^{KIP1}$ are not related with radiation induced-apoptosis.

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Characteristics of Non-Spherical Manganese Nodule from the East Siberian Sea (동시베리아해 비구형 망가니즈단괴의 특성)

  • Koo, HyoJin;Park, MuSeong;Seo, ChoongMan;Cho, HyenGoo
    • Korean Journal of Mineralogy and Petrology
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    • v.34 no.4
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    • pp.241-253
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    • 2021
  • Manganese nodules have been found in the shallow water depth of the Arctic Ocean as well as in the abyssal plains of the Pacific and Indian Oceans, but detailed study for them were rarely investigated. Manganese nodules, collected from the East Siberian Sea through the Arctic Expedition using Araon ice braking vessel, have a high potential for Mn mineral resources because they have high Mn content with high Mn/Fe ratio. This study investigated the external form, size and weight, internal texture for the non-spherical manganese nodule, which has about 7 % of total nodule from the East Siberian Sea. This study also researched the relative Mn-oxide mineral composition using the peak area ratio of X-ray diffraction pattern and their chemical composition. All data obtained from non-spherical nodules were compared with the spherical ones. Ellipsoidal, platy and irregular types are common among 5 groups of non-spherical manganese nodule based on the external form, and major axis and weight have positive relationship. All non-spherical manganese nodules have core mainly composed of mud sediments. The average Mn oxide mineral contents in nodules are birnessite, buserite and todorokite in descending order. Although mineral composition does not show any correlation with the external form, kind of core or internal structure, todorokite and buserite contents tend to increase and birnessite content decrease from the surface to the core in the nodule. Non-spherical manganese nodules have higher Mn content and Mn/Fe ratio than those from the shallow water depth of the Arctic Sea and even in the deep-sea of the Pacific and Indian Ocean. Although non-spherical nodule is larger and heavier, and has lower Mn content and Mn/Fe ratio than spherical nodule, there are not any differences in mineral composition and internal structure between them. Almost all manganese nodules collected from the East Siberian Sea are attributed to diagenetic process, because they are higher than 5 in Mn/Fe ratio.