• Title/Summary/Keyword: Large-cell

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Biological Functions of N- and O-linked Oligosaccharides of Equine Chorionic Gonadotropin and Lutropin/Chorionic Gonadotropin Receptor

  • Min, K.S.
    • Korean Journal of Animal Reproduction
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    • v.24 no.4
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    • pp.357-364
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    • 2000
  • Members of the glycoprotein family, which includes CG, LH, FSH and TSH, comprise two noncovalently linked $\alpha$- and $\beta$-subunits. Equine chorionic gonadotropin (eCG), known as PMSG, has a number of interesting and unique characteristics since it appears to be a single molecule that possesses both LH- and FSH-like activities in other species than the horse. This dual activity of eCG in heterologous species is of fundamental interest to the study of the structure-function relationships of gonadotropins and their receptors. CG and LH $\beta$ genes are different in primates. In horse, however, a single gene encodes both eCG and eLH $\beta$ -subunits. The subunit mRNA levels seem to be independently regulated and their imbalance may account for differences in the quantities of $\alpha$ - and $\beta$-subunits in the placenta and pituitary. The dual activities of eCG could be separated by removal of the N-linked oligosaccharide on the $\alpha$-subunit Asn 56 or CTP-associated O-linked oligosaccharides. The tethered-eCG was efficiently secreted and showed similar LH-like activity to the dimeric eCG. Interestingly, the FSH-like activity of the tethered-eCG was increased markedly in comparison with the native and wild type eCG. These results also suggest that this molecular can implay particular models of FSH-like activity not LH-like activity in the eCG/indicate that the constructs of tethered molecule will be useful in the study of mutants that affect subunit association and/or secretion. A single-chain analog can also be constructed to include additional hormone-specific bioactive generating potentially efficacious compounds that have only FSH-like activity. The LH/CG receptor (LH/CGR), a membrane glycoprotein that is present on testicular Leydig cells and ovarian theca, granulosa, luteal, and interstitial cells, plays a pivotal role in the regulation of gonadal development and function in males as well as in nonpregnant and pregnant females. The LH/CGR is a member of the family of G protein-coupled receptors and its structure is predicted to of a large extracellular domain connected to a bundle of seven membrane-spanning a-helices. The LH/CGR phosphorylation can be induced with a phorbol ester, but not with a calcium ionophore. The truncated form of LHR also was down-regulated normally in response to hCG stimulation. In contrast, the cell lines expressing LHR-t631 or LHR-628, the two phosphorylation-negative receptor mutant, showed a delay in the early phase of hCG-induced desensitization, a complete loss of PMA-induced desensitization, and an increase in the rate of hCG-induced receptor down-regulation. These results clearly show that residues 632~653 in the C-terminal tail of the LHR are involved in PMA-induced desensitization, hCG-induced desensitization, and hCG-induced down-regulation. Recently, constitutively activating mutations of the receptor have been identified that are associated with familial male-precocious puberty. Cells expressing LHR-D556Y bind hCG with normal affinity, exhibit a 25-fold increase in basal cAMP and respond to hCG with a normal increase in cAMP accumulation. This mutation enhances the internalization of the free and agoinst-occupied receptors ~2- and ~17- fold, respectively. We conclude that the state of activation of the LHR can modulate its basal and/or agonist-stimulated internalization. Since the internalization of hCG is involved in the termination of hCG actions, we suggest that the lack of responsiveness detected in cells expressing LHR-L435R is due to the fast rate of internalization of the bound hCG. This statement is supported by the finding that hCG responsiveness is restored when the cells are lysed and signal transduction is measured in a subcellular fraction (membranes) that cannot internalize the bound hormone.

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Intracullular Functions of the mas2+ Gene in the Fission Yeast, Schizosaccharomyces pombe (분열형 효모에서의 mas2+ 유전자의 세포 내 기능)

  • Sin, Sang-Min;Cha, Jae-Young;Ha, Se-Eun;Sim, Sun-Mi;Kim, Hyoung-Do;Lee, Jung-Sup;Park, Jong-Kun
    • Journal of Life Science
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    • v.19 no.1
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    • pp.101-110
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    • 2009
  • The regulation of gene expression plays an important role in cell cycle controls. In this study, a novel $mas2^+$ (mitosis associated protein) gene, a homolog of human SMARCAD1 was isolated and characterized from a fission yeast Schizosaccharomyces pombe (S. pombe) using gene-specific polymerase chain reaction. The isolated gene contained a complete open reading frame capable of encoding 922 amino acid residues with a typical promoter, as judged by nucleotide sequence analysis. It was also found that an SNF2 domain is located, which is involved in the chromosome remodeling. The quantitative analysis of the $mas2^+$ transcript against $adh1^+$ showed that the expression level of $mas2^+$ is high before septum formation in S. pombe. When $mas2^+$ null mutant cells were grown at 27 and $35^{\circ}C$, the cytokinesis of $mas2^+$ null mutant was greatly delayed and a large number of multi-septate and mis-segregated cells were produced. In addition, the number of multi-septate cells significantly increased. When cells were cultured in YES rich medium to increase proliferation, the abnormal phenotypes $mas2^+$ null mutant dramatically increased. These phenotypes could be rescued by an over-expression of the mast gene. The Mas2 protein localized in the nuclei of S. pombe, as evidenced by Mas2-EGFP signals. These results suggest that the $mas2^+$ is homologous to human SMARCAD1 gene and involved in septum formation and chromosome remodeling control.

Studies on the Ginseng Plants(III) -Radioactive Sodium $Acetate-U-C^{14}$ Feeding Experiments- (인삼식물(人參植物)에 관한 연구(III) -동위원소화합물(洞位元素化合物) Sodium $Acetate-U-C^{14}$을 투여한 실험-)

  • Kim, Jung-Yun;Staba, E. John
    • Korean Journal of Pharmacognosy
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    • v.5 no.2
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    • pp.111-124
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    • 1974
  • The radioactive compound sodium $acetate-U-C^{14}\;(C^{14}-acetate)$ was administered to two- and four-year-old July and September American ginseng (Araliaceae, Panax quinquefolium L.) plants and cuttings. The $C^{14}-acetate$ uptake was approximately 99%. The autoradiochromatograms suggest that the saponins isolated by preparative thin-layer chromatography contained impurities, especially those isolated from the leaf and stem extracts. The root and fruit methanol extracts yielded relatively pure saponins. The large amounts of panaquilin B and its proximity to panaquilin C on preparative thin-layer plates resulted in some admixing. The average concentration (% plant dry weight) of semi-purified saponins were high in the leaves (13.8%), as compared to fruits (9.8%), stems (7.9%) and roots (6.3%). The average percentage of $C^{14}-acetate$ incorporation into panaquilins was 4.8%. The average percentage of $C^{14}-acetate$ incorporation into panaquilins B and C was higher (1.40% and 1.13%, respectively) than that into panaquilins C, (d), G-1 and G-2 (0.75%, 0.65%, 0.13% and 0.53%, respectively). Panaquilin synthesis may be depending upon the part, collection period and age of the plant. The average percentage of $C^{14}-acetate$ incorporation into panaquilin B is high in roots (0.58%) and stems (0.48%); that into panaquilins C and (d) high in leaves (0.40% and 0.45%, respectively); and that into panaquilin E high in roots and leaves (0.55% and 0.50%, respectively). Panaquilin G-2 was synthesized in all parts of plants. The panaquilins appear to be biosynthesized more actively in July than September (exception-panaquilin G-1). Panaquilins B, C and G-1 may be biosynthesized more actively in four-year-old plants and panaquilins (d) and E more actively in two-year-old plants. The results from expectance with cuttings suggest that the panaquilins are synthesized de novo in the above-ground parts of ginseng plants, and that panaquilin G-1 may be synthesized de novo in the leaf. It is known from the tissue culture studies that panaquilins are produced by leaf, stem and root callus tissues and cailus-root cultures of American and Korean ginseng plants. Panaquilins may actively be synthesized de novo in most any cell or organ of the ginseng plants. It was verified that $C^{14}-acetate$ was incorporated into the panaxadiol portions of the panaquilins of two-year-old plants (sp. act. 0.56 mmcCi/mg) and four-year-old plants $(sp.\;act.\;0.54\;m{\mu}Ci/mg)$.

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Efficient of Hepatobiliary Scintigraphy both Decubitus Position in Biliary Leakage Patients (간담도 스캔 시 담즙 누출(Biliary Leakage)환자에서의 양측와위 자세(Both Decubitus Position)의 유용성)

  • Bahn, Young-Kag;Roh, Dong-Ook;Kang, Chun-Koo;Kim, Jae-Sam;Lee, Chang-Ho
    • The Korean Journal of Nuclear Medicine Technology
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    • v.12 no.3
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    • pp.229-234
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    • 2008
  • Hepatobiliary scintigraphy is very sensitivity of hepatic cell and gallbladder, biliary track atresia and biliary leakage. however, Hepatobiliary scan of biliary leakage diagnosis was separated determine biliary leakage and bowl drainage bile-juice. The object of this study will determine biliary leakage and bowl drainage bile-juice to hepatobiliary scintigraphy both decubitus position in bile leakage patients. Material & Methode: 31 patients (meal 14, Femeal 17), $51.1{\pm}14.4$ years. dynamic scan acquisition 60 farme for 60 minute on supine position. and delay scan was 2 hrs, 4 hrs, 24 hrs for 5 minute on supine, both decubitus position. Both decubitus position scan was kept for 5 minutes. Efficient of Hepatobiliary Scintigraphy both decubitus position in bile leakage patients was compared leakage size, density, image of supine position and both decubitus position. Results: 23 patients for 31 bile leakage patients was checked up function image or delay image, and 8 patients was checked up bile leakage on both decubitus. anatomical leakage location was supine position very well, but both decubitus position was separated bile leakage and moving bile-juice in bowl. also, uptake (counts/pixel) average of roi and bkg was supine 5.02, left decubitus 2.08, right decubitus 2.68. No. pixels of supine ROI counted 1.91 times than left decubitus, 1.05 times than right decubitus. Conclusion: 31 patient both decubitus position, but decubitus position was separated bile juice movement in bowl leakage location. also, It was compared ROI/BKG ratio and ROI No. pixels of supine, both decubitus in 38.5% patients. And No. pixels of supine position was large 19%, 5% than left decubitus, right decubitus, And density was in low 60%, 50% than left decubitus, right decubitus. It was mean bile leakage of ROI. so, If Hepatobiliary Scintigraphy was additional both decubitus position scan in bile leakage patients, this study will be more valuable in diagnosis of bile leakage.

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A Study on the Precise End-Point Detection in Titration by Using the Phase Angle Measurements (위상각 측정에 의한 적정의 정확한 종말점 검출법에 관한 연구)

  • Park, Byung-Bin;Shin, Ho-Sang;Lee, Han-Hyoung
    • Analytical Science and Technology
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    • v.12 no.4
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    • pp.290-298
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    • 1999
  • A study on the application of impedance phase angle for redox titration, acid-base titration, chelate titration and precipitation titration has been carried out. A constant alternating current was passed between two platinum electrodes. One of them was a polarizable micro-electrode of $0.1cm^2$ or $0.026cm^2$ surface area and the other a non-polarizable large electrode of $1cm^2$ surface area dipped in the solution to be titrated. The impedance and the phase angle of the titration cell were measured with lock-in amplifier to obtain well behaved titration curve respectively. In titration of oxalic acid vs. potassium permanganate, the end-point was obtained successfully from the phase angle titration curve. In this experiment, the concentration of 0.0005 M to 0.05 M, the current of $50{\mu}A$ and the frequency of near 50 Hz were used. In titration of phosphoric acid vs. sodium hydroxide, the first end-point was obtained successfully on the optimum experimental condition of 0.001 M concentration, $50{\mu}A$ current and 25~97 Hz frequency. However, the end-point in titration of cupric sulfate vs. disodium-EDTA couldn't be obtained clearly. The end-point was obtained with the out-of-phase impedance curve on the experimental condition of 0.01 M concentration, $100{\mu}A$ current, 5~35 Hz frequency range. In titration of sodium chloride vs. silver nitrate, the end-point was obtained successfully on the experimental condition of 0.1 M concentration, $100{\mu}A$ current and 5~47 Hz frequency range. This study showed that the impedance phase angle was applicable for the detection of the end-points in redox titration curve, acid-base titration curve, chelate titration curve and precipitation titration curve.

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Detection of Campylobacter jejuni in food and poultry visors using immunomagnetic separation and microtitre hybridization

  • Simard, Ronald-E.
    • Proceedings of the Korean Society of Fisheries Technology Conference
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    • 2000.05a
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    • pp.71-73
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    • 2000
  • Campylobacter jejuni is most frequently identified cause of cause of acute diarrhoeal infections in developeed countries, exceeding rates of illness caused by both salmonella and shigilla(Skirrow, 1990 ; Lior 1994). Previous studies on campylobacter jejuni contamination of commercial broiler carcasses in u.s.(Stern, 1992). Most cases of the disease result from indirect transmission of Campylobactor from animals via milk, water and meat. In addition to Campylobactor jejuni. the closely relates species Campylobactor coli and Campylobactor lari have also been implicated as agents of gastroenteritis in humans. Campylobactor coli represented only approximately 3% of the Campylobactor isolates from patients with Campylobactor enteritis(Griffiths and Park, 1990) whereas Campylobactor coli is mainly isolated from pork(Lmmerding et al., 1988). Campylobactor jejuni has also been isolated from cases of bacteremia, appendicitis and, recently, has been associated with Guillai-Barre syndrome(Allos and Blaser, 1994; von Wulffen et al., 1994; Phillips, 1995). Studies in volunteers indicated that the infectious dose for Campylobactor jejuni is low(about 500 organisms)(Robinson, 1981). The methods traditionally used to detect Campylobactor ssp. in food require at least two days of incubation in an enrichment broth followed by plating and two days of incubation on complex culture media containing many antibiotics(Goossens and Butzler, 1992). Finnaly, several biochemical tests must be done to confirm the indentification at the species level. Therfore, sensitive and specific methods for the detection of small numbers of Campylobactor cells in food are needed. Polymerase chain reaction(PCR) assays targeting specific DNA sequences have been developed for the detection of Campylobactor(Giesendorf and Quint, 1995; Hemandex et al., 1995; Winter and Slavidk, 1995). In most cases, a short enrichment step is needed to enhance the sensitivity of the assay prior to detection by PCR as the number of bacteria in the food products is low in comparison with those found in dinical samples, and because the complex composition of food matrices can hinder the PCR and lower its sensitivity. However, these PCR systems are technically demanding to carry out and cumbersome when processing a large number of samples simutaneously. In this paper, an immunomagnetic method to concentrate Campylobactor cells present in food or clinical samples after an enrichment step is described. To detect specifically the thermophilic Campylobactor. a monoclonal antibody was adsorbed on the surface of the magnetic beads which react against a major porin of 45kDa present on the surface of the cells(Huyer et al., 1986). After this partial purification and concentration step, detection of bound cells was achieved using a simple, inexpensive microtitre plate-based hybridization system. We examined two alternative detection systems, one specific for thermophilic Campylobactor based on the detection of 23S rRNA using an immobilized DNA probe. The second system is less specific but more sensitive because of the high copy number of the rRNA present in bacterial cell($10^3-10^4$). By using specific immunomagnetic beads against thermophilic Campylobactor, it was possible to concentrate these cells from a heterogeneous media and obtain highly specific hybridization reactions with good sensitivity. There are several advantages in using microtitre plates instead of filter membranes or other matrices for hybridization techniques. Microtitre plates are much easier to handle than filter membranes during the adsorption, washing, hybridization and detection steps, and their use faciilitates the simultanuous analysis of multiple sample. Here we report on the use of a very simple detection procedure based on a monoclonal anti-RNA-DNA hybrid antibody(Fliss et al., 1999) for detection of the RNA-DNA hybrids formed in the wells.

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Effect of Resistance Training on Skeletal Muscle Gene Expression in Rats: a Beadarray Analysis (저항성 운동이 골격근 유전자 발현에 미치는 영향: Beadarray 분석)

  • Oh, Seung-Lyul;Oh, Sang-Duk
    • Journal of Life Science
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    • v.23 no.1
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    • pp.116-124
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    • 2013
  • The aim was to examine resistance exercise-related genes after 8 weeks of resistance training. Thirty-two male Sprague-Dawley rats were divided into four groups: 4 weeks sedentary (4 wks CON, n=8), 8 weeks sedentary (8 wks CON, n=8), 4 weeks exercise training (4 wks REG, n=8), and 8 weeks exercise training (8 wks REG, n=8). The rats were trained to climb a 1-m vertical incline (85-degree), with weights secured to their tails. They climbed 10 times, 3 days per week, for 8 consecutive weeks. Skeletal muscle was taken from the flexor halucis longus after the exercise training. After separating the total RNA, large-scale gene expression was investigated by beadarray (Illumina RatRef-12 Expression BeadChip) analysis, and qPCR was used to inspect the beadarray data and to analyze the RNA quantitatively. The detection p-value for the genes was p<0.01, the M-value {M=$log_2$(condition)-$log_2$(reference)} was >1.0, and the DiffScore was >20. In total, the expression of 30 genes significantly increased 4 weeks after the exercise training, and the expression of six genes decreased. At 8 weeks, the expression of five genes significantly increased and that of 12 decreased. Several genes are potentially involved in resistance exercise and muscle hypertrophy, including 1) regulation of cell growth (IGFBP1, PLA2G2A, OKL38); 2) myogenesis (CSRP3); 3) tissue regeneration and muscle development (MUSTN1, MYBPH); 4) hypertrophy (CYR61, ATF3, NR4A3); and 5) glucose metabolism (G6PC, PCK1). These results may help to explain previously reported physiological changes of the skeletal muscle and suggest new avenues for further investigation.

Application of Transposable Elements as Molecular-marker for Cancer Diagnosis (암 진단 분자 마커로서 이동성 유전인자의 응용)

  • Kim, Hyemin;Gim, Jeong-An;Woo, Hyojeong;Hong, Jeonghyeon;Kim, Jinyeop;Kim, Heui-Soo
    • Journal of Life Science
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    • v.27 no.10
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    • pp.1215-1224
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    • 2017
  • Until now, various oncogenic pathways were idenfied. The accumulation of DNA mutation induces genomic instability in the cell, and it makes cancer. The development of bioinformatics and genomics, to find the precise and reliable biomarker is available. This biomarker could be applied the early-dignosis, prediction and convalescence of cancer. Recently, Transposable elements (TEs) have been attracted as the regulator of genes, because they occupy a half of human genome, and the cause of various diseases. TEs induce DNA mutation, as well as the regulation of gene expression, that makes to cancer development. So, we confirmed the relationship between TEs and colon cancer, and provided the clue for colon cancer biomarker. First, we confirmed long interspersed nuclear element-1 (LINE-1), Alu, and long terminal repeats (LTRs) and their relationship to colon cancer. Because these elements have large composition and enormous effect to the human genome. Interestingly, colon cancer specific patterns were detected, such as the hypomethylation of LINE-1, LINE-1 insertion in the APC gene, hypo- or hypermethylation of Alu, and isoform derived from LTR insertion. Moreover, hypomethylation of LINE-1 in proto-oncogene is used as the biomarker of colon cancer metastasis, and MLH1 mutation induced by Alu is detected in familial or hereditary colon cancer. The genes, effected by TEs, were analyzed their expression patterns by in silico analysis. Then, we provided tissue- and gender-specific expression patterns. This information can provide reliable cancer biomarker, and apply to prediction and diagnosis of colon cancer.

Comparison of Size Criteria in Mediastinal Lymph Node Involvement of Adenocarcinoma of Lungs (폐 선암의 종격동 림프절 전이에 있어서 림프절 크기 기준의 비교)

  • Gu, Ki-Seon;Kuk, Hiang;Koh, Hyeck-Jae;Yang, Sei-Hun;Jeong, Eun-Taik
    • Tuberculosis and Respiratory Diseases
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    • v.46 no.4
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    • pp.542-547
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    • 1999
  • Background: Decision in mediastinal lymph node involvement of lung cancer by CT scan is very important and valuable for the treatment planning and prognosis prediction. In general, long diameter of mediastinal lymph node more than 15mm is used as criterion of lung cancer involvement. Adenocarci-noma has a tendency of early distant metastasis and micrometastasis, so adenocarcinoma may involve lymph node earlier and cannot be detected before lymph nodes are enlarged enough. The authors tried to determine the difference between two size criteria(15mm, 10mm) in adenocarcinoma for the detection of cancer involvement. Methods: Numbers of sample are 60 cases(male 46, female 14, median age: 61.5 years). According to pathology, squamous cancer 41, large cell cancer 2, adenocarcinoma 17. According to TNM stage, I 23, III 24, IIIA 13. Results : Mean long diameter of lymph node involvement is 16.0($\pm8.0$) mm in non-adenocarcinoma group, and that of adenocarcinoma group is 12.0($\pm3.2$) mm(p<0.05). If long diameter of lymph node larger than 15mm as involvement criterion is applied, sensitivity, specificity, positive predictive index, negative predictive index, accuracy of nonadenocarcinoma group are 54%, 100%, 100%, 83%, 86%, and those of adenocarcinoma group are 43%, 90%, 75%, 69%, 71%. If long diameter of lymph node larger than 10mm as involvement criterion is applied, sensitivity, specificity, positive predictive index. negative predictive index. accuracy of nonadenocarcinoma group are 65%, 77%, 61%, 92%, 79%, and those of adenocarcinoma group are 100%, 80%, 78%, 100%, 88%. Conclusion: Long diameter of lymph node larger than 10mm is more valuable criterion as lymph node involvement in adenocarcinoma of lungs.

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A Study on the Image Registration Algorithms for the Accurate Application of Multimodality Image in Radiation Treatment Planning (방사선치료 계획시 다중영상 활용의 정확도 향상을 위한 영상정합 알고리즘 분석)

  • 송주영;이형구;최보영;윤세철;서태석
    • Progress in Medical Physics
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    • v.13 no.4
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    • pp.209-217
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    • 2002
  • There have been many studies on the application of the reciprocal advantages of multimodality image to define accurate target volume in the Process of radiation treatment planning. For the proper use of the multimodality images, the registration works between different modality images should be performed in advance. In this study, we selected chamfer matching method and mutual information method as most popular methods in recent image registration studies considering the registration accuracy and clinical practicality. And the two registration methods were analyzed to deduce the optimal registration method according to the characteristics of images. Lung phantom of which multimodality images could be acquired was fabricated and CT, MRI and SPECT images of the phantom were used in this study. We developed the registration program which can perform the two registration methods properly and analyzed the registration results which were produced by the developed program in many different images' conditions. Although the overall accuracy of the registration in both chamfer matching method and mutual information method was acceptable, the registration errors in SPECT images which had lower resolution and in degraded images of which data were removed in some part were increased when chamfer matching method was applied. Especially in the case of degraded reference image, chamfer matching methods produce relatively large errors compared with mutual information method. Mutual information method can be estimated as more robust registration method than chamfer matching method in this study because it did not need the prerequisite works, the extraction of accurate contour points, and it produced more accurate registration results consistently regardless of the images' characteristics. The analysis of the registration methods in this study can be expected to provide useful information to the utilization of multimodality images in delineating target volume for radiation treatment planning and in many other clinical applications.

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