• 제목/요약/키워드: semi-quantitative analysis

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Reconstructed Adeno-Associated Virus with the Extracellular Domain of Murine PD-1 Induces Antitumor Immunity

  • Elhag, Osama A.O.;Hu, Xiao-Jing;Wen-Ying, Zhang;Li, Xiong;Yuan, Yong-Ze;Deng, Ling-Feng;Liu, De-Li;Liu, Ying-Le;Hui, Geng
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.8
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    • pp.4031-4036
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    • 2012
  • Background: The negative signaling provided by interactions of the co-inhibitory molecule, programmed death-1 (PD-1), and its ligands, B7-H1 (PD-L1) and B7-DC (PD-L2), is a critical mechanism contributing to tumor evasion; blockade of this pathway has been proven to enhance cytotoxic activity and mediate antitumor therapy. Here we evaluated the anti-tumor efficacy of AAV-mediated delivery of the extracellular domain of murine PD-1 (sPD-1) to a tumor site. Material and Methods: An rAAV vector was constructed in which the expression of sPD-1, a known negative regulator of TCR signals, is driven by human cytomegalovirus immediate early promoter (CMV-P), using a triple plasmid transfection system. Tumor-bearing mice were then treated with the AAV/sPD1 construct and expression of sPD-1 in tumor tissues was determined by semi quantitative RT-PCR, and tumor weights and cytotoxic activity of splenocytes were measured. Results: Analysis of tumor homogenates revealed sPD-1 mRNA to be significantly overexpressed in rAAV/sPD-1 treated mice as compared with control levels. Its use for local gene therapy at the inoculation site of H22 hepatoma cells could inhibit tumor growth, also enhancing lysis of tumor cells by lymphocytes stimulated specifically with an antigen. In addition, PD-1 was also found expressed on the surfaces of activated CD8+ T cells. Conclusion: This study confirmed that expression of the soluble extracellular domain of PD-1 molecule could reduce tumor microenvironment inhibitory effects on T cells and enhance cytotoxicity. This suggests that it might be a potential target for development of therapies to augment T-cell responses in patients with malignancies.

Analysis of Immune Response in Dogs with Chronic Inflammatory Skin Disease (만성 피부 염증소견을 보인 개의 면역학적 특성 분석 연구)

  • Cho, Sun-Ju;Koh, Min-Soo;Jung, Bock-Gie;Ko, Jae-Hyung;Yoon, So-Rah;Han, Dong-Un;Lee, Bong-Joo
    • Journal of Veterinary Clinics
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    • v.26 no.5
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    • pp.433-440
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    • 2009
  • High levels of inflammatory cytokines were proposed contributors to the pathogenesis of a various inflammatory skin disorders. Therefore, investigating the immune response of the inflammatory skin disorder allows a better understanding of pathogenesis of a various inflammatory skin disorders and therapeutic approaches. The aim of this study was to analyze of the immune response in dogs with chronic inflammatory skin disease. To this aim, the present study evaluated relative mRNA expression of canine $IFN-{\gamma}$, IL-4, $TGF-{\beta}$ and IL-10 using TaqMan realtime PCR assays and semi-quantitative RT-PCR in freshly isolated peripheral blood mononuclear cells from twenty dogs with chronic inflammatory skin disease and ten normal dogs. The relative mRNA expression levels of IL-4 mRNA were significantly higher in dogs with chronic inflammatory skin disease than those in normal dogs (P < 0.01). The results of present study also showed a tendency towards increased expression of IL-10 transcripts in dogs with chronic inflammatory skin disease. However, there were no significant differences in the levels $IFN-{\gamma},\;TGF-{\beta}$ between normal and chronically inflammed dogs. In addition, the concentration of serum IgE was significantly increased in dogs with chronic inflammatory skin disease compared with those in normal dogs (P < 0.01). In histopathological examination, we found that there were markedly increased mast cell counts in chronically inflammed dogs (P < 0.05). These results suggest that the pathogenesis of chronic inflammatory skin disease might be associated with a T-cell mediated inflammatory responses characterized by a Th2-skewed immune response. Based on these results, the modulation of Th1/Th2 balance may be an effective therapeutic strategy for the treatment of chronic inflammatory skin disease.

Theoretical Study on Structural Properties of Phenthylamine Derivatives (페네틸아민 유도체의 구조적 특성에 관한 이론적 연구)

  • Lee, Chul Jae
    • The Journal of the Convergence on Culture Technology
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    • v.6 no.4
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    • pp.761-766
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    • 2020
  • Phenylamine derivatives are substances that have a biochemical action and are widely applied as psychotropic drugs. In particular, with regard to the quantitative analysis of substances such as ephedrine, amphetamine, pentermine, and dopamine, many previous studies such as electrochemical, vacuum ultraviolet method, and gas chromatography have been conducted. However, there have not been many studies on the structural characteristics of molecular units. Therefore, In this study, we used (HyperChem8.0's, HC) semi-empirical PM3 method to calculate the total energy, band gap, electrostatic potential, and net charge of ephedrine, amphetamine, pentamine and dopamine to investigate the chemical properties of each derivative according to the molecular structure change. The results showed that for total energy -43,171.8, -32,9538.3, -36,407.3 and -43,061.2 Kcal/mol, respectively, while for band gaps, 10.16379377, 9.9531666, 9.7878002 and 9.0589282 eV. Also, for electrostatic potentials, 1.301~-0.045, 1.694~0.299, 0.694~-0.158 and 1.587~-0.048 respectively. Finally, looking at the distribution of net charges, the oxygen atoms, nitrogen atoms and carbon atoms were -0.312~-0.242, -0.161~-0.051 and +0.13~-0.12 respectively. These results are expected to lead to chemical action centered on phenyl radicals and oxygen and nitrogen atoms common to phenethylamine derivatives.

Simple and Rapid Evaluation System for Endosulfan Toxicity and Selection of Endosulfan Detoxifying Microorganism Based on Lumbricus rubellus (Lumbricus rubellus를 이용한 endosulfan의 간편, 신속 독성 평가 및 endosulfan 분해 미생물의 선별)

  • Sohn Ho-Yong;Kim Hong-Ju;Kum Eun-Joo;Lee Jung-Bok;Kwon Gi-Seok
    • Journal of Life Science
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    • v.16 no.1
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    • pp.108-113
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    • 2006
  • To compensate the problems of chemical assay in detoxification of recalcitrant and a practical approach in selection of bioremediation bacteria, a simple and rapid toxicity evaluation system was constructed based on Lumbricus rubellus. Long term-culture and specific equipment are not necessary, and semi-quantitative analysis of toxicity at sub-lethal concentration is possible by measuring of dose-dependent increased yellowish secreted compounds. When the toxicity of endosulfan, its metabolites and structurally related chemicals were measured for 24 h, the results were coincided with previous reports. Toxicity was found in endosulfan, endosulfan sulfate, aldrin, and dieldrin, respectively. Rapid and economic selection of endosulfan-detoxifying bacteria was possible using our system. Klebsiella pneumoniae KE-1, K. oxytoca KE-8 and Pseudomonas sp. KS-2P, reported endosulfan degrading bacteria, ameliorated the endosulfan toxicity, whereas E. coli, B. subtilis and other bacteria failed to protect the toxicity of endosulfan in L. rubellus. Our results suggest that the constructed system is useful to selection of microorganism as well as toxicity evaluation against toxic recalcitrants.

Development of a Food Frequency Questionnaire for the Korea National Health and Nutrition Examination Survey: Data from the Fourth Korea National Health and Nutrition Examination Survey (KNHANES IV) (국민건강영양조사 식품섭취빈도조사지 개발: 국민건강영양조사 제4기 (2007-2009) 자료 이용)

  • Yun, Sung Ha;Shim, Jee-Seon;Kweon, Sanghui;Oh, Kyungwon
    • Journal of Nutrition and Health
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    • v.46 no.2
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    • pp.186-196
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    • 2013
  • The objective of this study was to develop a semi-quantitative food frequency questionnaire (FFQ) for an adult population for use in the Korea National Health and Nutrition Examination Survey (KNHANES). The 24-hour recall data for 2,939 subjects aged 19 years and over from the fourth KNHANES first year (2007) were used to extract the items included in the questionnaire. The FFQ items were developed by selection of major dishes based on the cumulative contribution rate, between-person variability based on the cumulative R2 of energy and 14 nutrients (carbohydrate, protein, fat, crude fiber, calcium, phosphorous, iron, sodium, potassium, vitamin A, thiamin, riboflavin, niacin, and vitamin C) and the number of consumers of each dish. In addition, the FFQ items were revised with analysis of 24-hour recall data of the fourth KNHANES second and third year (2008, 2009). Finally, 112 items were included in the FFQ and grouped as follows: rice (5 items), noodles and dumplings (6), breads and rice cakes (8), soups and stews (12), soybeans, eggs, meat and fish (23), vegetables, seaweed and potatoes (27), milk and dairy products (4), fruits (13), beverages (5), snacks (6) and alcoholic beverages (3). The food items of FFQ accounted for an average of 87.0% of energy and 14 nutrient intakes and also accounted for 81.7% of the between-person variability. The frequency range of the FFQ items was classified into nine categories (never or seldom, once per month, 2-3 times per month, once per week, 2-4 times per week, 5-6 times per week, once per day, twice per day and three times per day) and the portion size was divided into three categories (small, medium and large). We expect that this developed dish-based FFQ could be used in assessment of longterm dietary intakes of Korean adults.

Development of Cloud-Based Medical Image Labeling System and It's Quantitative Analysis of Sarcopenia (클라우드기반 의료영상 라벨링 시스템 개발 및 근감소증 정량 분석)

  • Lee, Chung-Sub;Lim, Dong-Wook;Kim, Ji-Eon;Noh, Si-Hyeong;Yu, Yeong-Ju;Kim, Tae-Hoon;Yoon, Kwon-Ha;Jeong, Chang-Won
    • KIPS Transactions on Computer and Communication Systems
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    • v.11 no.7
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    • pp.233-240
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    • 2022
  • Most of the recent AI researches has focused on developing AI models. However, recently, artificial intelligence research has gradually changed from model-centric to data-centric, and the importance of learning data is getting a lot of attention based on this trend. However, it takes a lot of time and effort because the preparation of learning data takes up a significant part of the entire process, and the generation of labeling data also differs depending on the purpose of development. Therefore, it is need to develop a tool with various labeling functions to solve the existing unmetneeds. In this paper, we describe a labeling system for creating precise and fast labeling data of medical images. To implement this, a semi-automatic method using Back Projection, Grabcut techniques and an automatic method predicted through a machine learning model were implemented. We not only showed the advantage of running time for the generation of labeling data of the proposed system, but also showed superiority through comparative evaluation of accuracy. In addition, by analyzing the image data set of about 1,000 patients, meaningful diagnostic indexes were presented for men and women in the diagnosis of sarcopenia.

Development of Gated Myocardial SPECT Analysis Software and Evaluation of Left Ventricular Contraction Function (게이트 심근 SPECT 분석 소프트웨어의 개발과 좌심실 수축 기능 평가)

  • Lee, Byeong-Il;Lee, Dong-Soo;Lee, Jae-Sung;Chung, June-Key;Lee, Myung-Chul;Choi, Heung-Kook
    • The Korean Journal of Nuclear Medicine
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    • v.37 no.2
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    • pp.73-82
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    • 2003
  • Objectives: A new software (Cardiac SPECT Analyzer: CSA) was developed for quantification of volumes and election fraction on gated myocardial SPECT. Volumes and ejection fraction by CSA were validated by comparing with those quantified by Quantitative Gated SPECT (QGS) software. Materials and Methods: Gated myocardial SPECT was peformed in 40 patients with ejection fraction from 15% to 85%. In 26 patients, gated myocardial SPECT was acquired again with the patients in situ. A cylinder model was used to eliminate noise semi-automatically and profile data was extracted using Gaussian fitting after smoothing. The boundary points of endo- and epicardium were found using an iterative learning algorithm. Enddiastolic (EDV) and endsystolic volumes (ESV) and election fraction (EF) were calculated. These values were compared with those calculated by QGS and the same gated SPECT data was repeatedly quantified by CSA and variation of the values on sequential measurements of the same patients on the repeated acquisition. Results: From the 40 patient data, EF, EDV and ESV by CSA were correlated with those by QGS with the correlation coefficients of 0.97, 0.92, 0.96. Two standard deviation (SD) of EF on Bland Altman plot was 10.1%. Repeated measurements of EF, EDV, and ESV by CSA were correlated with each other with the coefficients of 0.96, 0.99, and 0.99 for EF, EDV and ESV respectively. On repeated acquisition, reproducibility was also excellent with correlation coefficients of 0.89, 0.97, 0.98, and coefficient of variation of 8.2%, 5.4mL, 8.5mL and 2SD of 10.6%, 21.2mL, and 16.4mL on Bland Altman plot for EF, EDV and ESV. Conclusion: We developed the software of CSA for quantification of volumes and ejection fraction on gated myocardial SPECT. Volumes and ejection fraction quantified using this software was found valid for its correctness and precision.

QTL Analysis to Improve and Diversify the Grain Shape of Rice Cultivars in Korea, Using the Long Grain japonica Cultivar, Langi (초장립종 벼를 이용한 입형 관련 QTL 분석 및 국내 벼 품종 입형 개선 연구)

  • Kim, Suk-Man;Park, Hyun-Su;Lee, Chang-Min;Baek, Man-Kee;Cho, Young-Chan;Suh, Jung-Pil;Jeong, Oh-Young
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.65 no.4
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    • pp.303-313
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    • 2020
  • Rice grain shape is one of the key components of grain yield and market value. An understanding of the genetic basis of the variation in grain shape could be used to improve grain shape. In this study, we developed a total of 265 F2 individuals derived from a cross between japonica cultivars (Josaeng-jado and Langi) and used this population for quantitative trait locus (QLT) analysis. Correlation analysis was performed to identify relationships between grain traits (GL: grain length, GW: grain width, L/W: ratio of length to width, TGW: 1,000 grain weight). The grain shape was positively correlated with GL and TGW, and negatively correlated with GW. In QTL analysis associated with grain shape, one QTL for GL, qGL5, detected on chromosome 5, explained 20.3% of the phenotypic variation (PV), while two QTLs, qGW5 (PV=36.1) and qGW7 (PV=26.1), for GW were identified on chromosomes 5 and 7, respectively. Evaluation of the effects of each of the QTLs on the grain shape in the population showed a significant difference in the grain size in positive lines compared with the lines without the QTLs. According to the QTL combination of the allelic-types, the grain shape of the tested lines varied from semi-round type to long spindle-shaped type. The results of this study extend our knowledge about the genetic pool governing the diversity of grain shape in japonica cultivars and could be used to improve the grain shape of this species through marker-assisted selective breeding in Korea.

The Effect of Nonspecific Endothelin-1 Receptor Blocker ($Bosentan^{(R)}$) on Paraquat Induced Pulmonary Fibrosis in Rat (Paraquat에 의한 백서의 폐섬유화증에서 비선택적 Endothelin-1 receptor blocker($Bosentan^{(R)}$)의 치료효과)

  • Jeong, Hye-Cheol;Jung, Ki-Hwan;Kim, Byung-Gyu;Lee, Seung-Heon;Kim, Min-Kyung;Kim, Chung-Yeul;Park, Sang-Myun;Lee, Sin-Hyung;Shin, Chol;Cho, Jae-Youn;Shim, Jae-Jeong;In, Kwang-Ho;Kim, Han-Gyum;Yoo, Se-Hwa;Kang, Kyung-Ho
    • Tuberculosis and Respiratory Diseases
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    • v.50 no.2
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    • pp.182-195
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    • 2001
  • Background : Idiopathic pulmonary fibrosis(IPF) is a devastating illness for which there is little effective treatment. The key cytokines currently implicated in the fibrotic process are the transforming growth factor-${\beta}_1$(TGF-${\beta}_1$), tumor necrosis factor-$\alpha$(TNF-$\alpha$), endothelin-1(ET-1) and interferon-$\gamma$(IFN-$\gamma$). The rat model for paraquat-induced pulmonary fibrosis was chosen to investigate the role of ET-1 in this disease. Both ET-1 and TGF-${\beta}_1$ expression in lung lesions were examined using immunohistochemical staining. After $Bosentan^{(R)}$ administration, an orally active ET-$l_A$ and ET-$1_B$ receptor antagonist, the degree of pulmonary fibrosis and ET-1 and TGF-${\beta}_1$ expression were analyzed. Method : Sprague-Dawley rats were divided into three groups, the control group, the fibrosis group, and the fibrosis-$Bosentan^{(R)}$-treated group. The animals were sacrificed periodically at 1, 3, 5, 7, 10, 14 days after administering saline or paraquat. The effects between groups were compared with the results of light microscopy and immunohistochemical staining for ET-1 and TGF-${\beta}_1$. The degree of fibrosis was evaluated by H&E and Masson's trichrome staining, which were graded by a computerized image analyzer. The degree of immunohistochemical staining was categorized by a semi-quantitative analysis method. Results : The lung collagen content had increased in the paraquat instillated animals by day 3, and continued to increase up to day 14. A daily treatment by gavage with $Bosentan^{(R)}$ (100mg/kg) did not prevent the increase in collagen deposition on the lung that was induced by paraquat instillation. There were increased immunohistochemical stains of ET-1 on the exudate, macrophages, vascular endothelial cells and pneumocytes in the paraquat instillated group. Furthermore, TGF-${\beta}_1$ expression was higher on the exudate, macrophages, some inflammatory cells, pneumocytes( type I, and II), vascular endothelium and the respiratory epithelial cells around the fibrotic area. After Bosentan treatment, there were no definite changes in ET-1 and TGF-${\beta}_1$ expression. Conclusion : Fibrosis of the Paraquat instillated group was more advanced when compared with the control group. In addition, there was increased ET-1 and TGF-${\beta}_1$ expression around the fibrotic area. ET-1 is associated with lung fibrosis but there was little effect of the ET-1 receptor blocker($Bosentan^{(R)}$) on antifibrosis.Background : Idiopathic pulmonary fibrosis(IPF) is a devastating illness for which there is little effective treatment. The key cytokines currently implicated in the fibrotic process are the transforming growth factor-${\beta}_1$(TGF-${\beta}_1$), tumor necrosis factor-$\alpha$(TNF-$\alpha$), endothelin-1(ET-1) and interferon-$\gamma$(IFN-$\gamma$). The rat model for paraquat-induced pulmonary fibrosis was chosen to investigate the role of ET-1 in this disease. Both ET-1 and TGF-${\beta}_1$ expression in lung lesions were examined using immunohistochemical staining. After $Bosentan^{(R)}$ administration, an orally active ET-$1_A$ and ET-$1_B$ receptor antagonist, the degree of pulmonary fibrosis and ET-1 and TGF-${\beta}_1$ expression were analyzed. Method : Sprague-Dawley rats were divided into three groups, the control group, the fibrosis group, and the fibrosis-$Bosentan^{(R)}$-treated group. The animals were sacrificed periodically at 1, 3, 5, 7, 10, 14 days after administering saline or paraquat. The effects between groups were compared with the results of light microscopy and immunohistochemical staining for ET-1 and TGF-${\beta}_1$. The degree of fibrosis was evaluated by H&E and Masson's trichrome staining, which were graded by a computerized image analyzer. The degree of immunohistochemical staining was categorized by a semi-quantitative analysis method. Results : The lung collagen content had increased in the paraquat instillated animals by day 3, and continued to increase up to day 14. A daily treatment by gavage with $Bosentan^{(R)}$ (100mg/kg) did not prevent the increase in collagen deposition on the lung that was induced by paraquat instillation. There were increased immunohistochemical stains of ET-1 on the exudate, macrophages, vascular endothelial cells and pneumocytes in the paraquat instillated group. Furthermore, TGF-${\beta}_1$ expression was higher on the exudate, macrophages, some inflammatory cells, pneumocytes( type I, and II), vascular endothelium and the respiratory epithelial cells around the fibrotic area. After Bosentan treatment, there were no definite changes in ET-1 and TGF-${\beta}_1$ expression. Conclusion : Fibrosis of the Paraquat instillated group was more advanced when compared with the control group. In addition, there was increased ET-1 and TGF-${\beta}_1$ expression around the fibrotic area. ET-1 is associated with lung fibrosis but there was little effect of the ET-1 receptor blocker($Bosentan^{(R)}$) on antifibrosis.

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Expression and Localization of ATF4 Gene on Oxidative Stress in Preimplantation Mouse Embryo (생쥐 착상전 배아에서 산화적 스트레스에 의한 ATF4 유전자의 발현과 존재 부위)

  • Na, Won-Heum;Kang, Han-Seung;Eo, Jin-Won;Gye, Myung-Chan;Kim, Moon-Kyoo
    • Development and Reproduction
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    • v.10 no.2
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    • pp.105-113
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    • 2006
  • Reactive oxygen species(ROS) generated in cellular metabolism have an effect on cell maturation and development. In human reproductive tract, oxidative injury by ROS may induce female infertility. Also, oxidative injury may be responsible for developmental retardation and arrest of mammalian preimplantation embryos. Activating transcription factor 4(ATF4) is a member of the cyclic-AMP response element-binding(CREB) familiy of basic region- leucine zipper(bZip). ATF4 is known to regulate stress response to protect cell from various stress factors and inducer of apoptisis. The purpose of this study was to investigate whether ATF4 is involved in the defensive mechanism in oxidative stress condition during the development of mouse preimplantation embryos. To verify the expression of ATF4 in oxidative stress condition, 2-cell stage embryos were cultured in HTF media containing 0.1mM, 0.5mM or 1mM hydrogen peroxide($H_2O_2$) for 1hr(2-cell), 8hr(4-cell), 17hr(8-cell), 24hr(morula), 48hr(early blastocyst) or 64hr(late blastocyst). The developmental rate decreased in the 0.1mM $H_2O_2$ treated group compared with control group. In embryos treated with 0.5mM and 1mM $H_2O_2$ showed 2-cell block. As a results of the semi-quantitative RT-PCR analysis of SOD1, ATF4 and Bax gene expression, SOD1, ATF4 and Bax genes were increased in 0.1mM, 0.5mM, 1mM $H_2O_2$ treated groups compared with control group. In 2-cell embryos, expression of SOD1, ATF4 and Bax genes were notably increased in 0.1mM, 0.5mM, 1mM $H_2O_2$ treated groups compared with control group. Immunofluorescence analysis showed that ATF4 protein was localized at the cytoplasm of preimplantation embryos. The increase in ATF4 immunoreactivety was observed in the 0.1mM, 0.5mM, 1mM $H_2O_2$ treated groups compared with control group. It suggests that oxidative stress by $H_2O_2$ induces expression of ATF4 and may be involved in protection mechanism in preimplantation embryos from oxidative injury.

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