• Title/Summary/Keyword: semi-quantitative analysis

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COMPARISON OF SALIVARY ANTIBODY IgA TITRE TO STREPTOCOCCUS MUTANS BETWEEN THE CARIES-EXPERIENCED AND NON-EXPERIENCED GROUPS USING IMMUNO-SLOT BLOT METHOD (Immuno-slot blot method를 이용한 우식 경험 아동과 비경험 아동간의 Streptococcus mutans에 대한 타액내 IgA 역가의 비교)

  • Eum, Jong-Hyeok;Jeong, Tae-Sung;Kim, Shin
    • Journal of the korean academy of Pediatric Dentistry
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    • v.29 no.3
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    • pp.354-361
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    • 2002
  • The aim of this study is to develop and establish rapid, convenient, and accurate method of analyzing salivary IgA against S. mutans semi-quantitatively. Relative salivary IgA titer was calculated as maximum dilution fold of S. mutans protein that was not detected by salivary antibody after measuring relative intensity of the immune blot bands by densitometry. Analyses were performed in caries-experienced and non-experienced children. Mean IgA titer of non-experienced group shows higher level than that of caries-experienced without statistical significance due to high individual variety of antibody titer in non-experienced group: $2^{6.278{\pm}2.260}$ in non-experienced group and $2^{5.730{\pm}0.499}$ in caries-experienced group(p=0.464). Those results suggest that naturally induced salivary IgA antibodies against S. mutans were present in all subjects, but high titer of antibodies were not achieved in caries-experienced group. On the contrary, antibody titer in non-experienced group shows marked individual variations suggesting that antibody production is multifactorial. In conclusion, immune-slot blot method developed in this study would be useful and applicable in semi-quantitative analysis of antibodies.

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The Effect of PET/CT Images on SUV with the Correction of CT Image by Using Contrast Media (PET/CT 영상에서 조영제를 이용한 CT 영상의 보정(Correction)에 따른 표준화섭취계수(SUV)의 영향)

  • Ahn, Sha-Ron;Park, Hoon-Hee;Park, Min-Soo;Lee, Seung-Jae;Oh, Shin-Hyun;Lim, Han-Sang;Kim, Jae-Sam;Lee, Chang-Ho
    • The Korean Journal of Nuclear Medicine Technology
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    • v.13 no.1
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    • pp.77-81
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    • 2009
  • Purpose: The PET of the PET/CT (Positron Emission Tomography/Computed Tomography) quantitatively shows the biological and chemical information of the body, but has limitation of presenting the clear anatomic structure. Thus combining the PET with CT, it is not only possible to offer the higher resolution but also effectively shorten the scanning time and reduce the noises by using CT data in attenuation correction. And because, at the CT scanning, the contrast media makes it easy to determine a exact range of the lesion and distinguish the normal organs, there is a certain increase in the use of it. However, in the case of using the contrast media, it affects semi-quantitative measures of the PET/CT images. In this study, therefore, we will be to establish the reliability of the SUV (Standardized Uptake Value) with CT data correction so that it can help more accurate diagnosis. Materials and Methods: In this experiment, a total of 30 people are targeted - age range: from 27 to 72, average age : 49.6 - and DSTe (General Electric Healthcare, Milwaukee, MI, USA) is used for equipment. $^{18}F$- FDG 370~555 MBq is injected into the subjects depending on their weight and, after about 60 minutes of their stable position, a whole-body scan is taken. The CT scan is set to 140 kV and 210 mA, and the injected amount of the contrast media is 2 cc per 1 kg of the patients' weight. With the raw data from the scan, we obtain a image showing the effect of the contrast media through the attenuation correction by both of the corrected and uncorrected CT data. Then we mark out ROI (Region of Interest) in each area to measure SUV and analyze the difference. Results: According to the analysis, the SUV is decreased in the liver and heart which have more bloodstream than the others, because of the contrast media correction. On the other hand, there is no difference in the lungs. Conclusions: Whereas the CT scan images with the contrast media from the PET/CT increase the contrast of the targeted region for the test so that it can improve efficiency of diagnosis, there occurred an increase of SUV, a semi-quantitative analytical method. In this research, we measure the variation of SUV through the correction of the influence of contrast media and compare the differences. As we revise the SUV which is increasing in the image with attenuation correction by using contrast media, we can expect anatomical images of high-resolution. Furthermore, it is considered that through this trusted semi-quantitative method, it will definitely enhance the diagnostic value.

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Effects of Melatonin on Gene Expression of IVM/IVF Porcine Embryos

  • Jang, H.Y.;Kong, H.S.;Choi, K.D.;Jeon, G.J.;Yang, B.K.;Lee, C.K.;Lee, H.K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.1
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    • pp.17-21
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    • 2005
  • The effect of melatonin on in vitro embryo development and the expression of antioxidant enzyme gene in preimplantation porcine embryos was determined by modified semi-quantitative single cell RT-PCR. Porcine embryos derived from in vitro maturation /in vitro fertilization were cultured in 5% $CO_2$ and 20% $O_2$ at $37^{\circ}C$ in NCSU23 medium. Melatonin was added to medium at concentration of 1nM, 5 nM, and 10 nM. When treated with 1nM (39.0%) of melatonin, the developmental rate of embryos beyond the morula stage were higher than that of control group (31.0%) (p<0.05). Number of inner cell mass and tropectoderm cell in control (23.0${\pm}$0.5 and 17.3${\pm}$0.8), 1 nM (23.6${\pm}$0.6 and 19.0${\pm}$0.5), and 5 nM (23.3${\pm}$1.1 and 16.3${\pm}$0.8) treated with melatonin were higher than in 10 nM (20.0${\pm}$0.5 and 13.3${\pm}$0.8) treated with melatonin (p<0.05). To develop an mRNA phenotypic map for the expression of catalase, bax and caspase-3, single cell RT-PCR analysis were carried out in porcine IVM/IVF embryo. Catalase was detected in 0, 1 and 5 nM supplemented with melatonin, but bax and caspase-3 were detected in 10 nM treated with melatonin.

Correlation between chromosome abnormalities and genomic imprinting in developing human - 1) Frequent biallelic expression of insulin-like growth factor II (IGF2) in gynogenetic Ovarian Teratomas: Uncoupling of H19 and IGF2 imprinting

  • Choi, Bo-Hwa;Lee, In-Hwan;Chun, Hyo-Jin;Kang, Shin-Sung;Chang, Sung-Ik
    • Journal of Genetic Medicine
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    • v.2 no.1
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    • pp.41-47
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    • 1998
  • Human uniparental gestations such as gynogenetic ovarian teratomas provide a model to evaluate the integrity of parent-specific gene expression - i.e. imprinting - in the absence of a complementary parental genetic contribution. The few imprinted genes characterized so far include the insulin-like growth factor-2 gene (IGF2) coding for a fetal growth factor and H19 gene whose normal function is unknown but it is likely to act as an mRNA. IGF2 is expressed by the paternal allele and H19 by the maternal allele. This reciprocal expression is quite interesting because both H19 and IGF2 genes are located close to each other on chromosome 11p15.5. In situ RNA hybridization analysis has shown variable expression of the H19 and IGF2 alleles according to the tissue origin in 11 teratomas. Especially, Skin, derivative of ectoderm, is expressed conspicuously. We examined imprinting of H19 and IGF2 in teratomas using PCR and RT-PCR of exonic polymorphism. H19 and IGF2 transcript could be expressed either biallelically or monoallelically in the teratomas. Biallelic expression (i.e., loss of imprinting) of IGF2 occurred in 5 out of 6 mature teratomas and 1 out of 1 immature teratoma. Biallelic expression of H19 occurred in 4 out of 10 mature teratomas and 1 out of 1 immature teratoma. Expression levels of H19 and IGF2 transcript using the semi-quantitative RT-PCR had no relation between monoallelic and biallelic expression. Moreover, IGF2 biallelic expression did not affect allele-specificity or levels of H19 expression. These results demonstrate that both genes, H19 and IGF2, can be imprinted, expressed and regulated independently and individually of each other in ovarian teratoma.

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Molecular Mechanism of Atopic Dermatitis Induction Following Sensitization and Challenge with 2,4-Dinitrochlorobenzene in Mouse Skin Tissue

  • Kim, JiYoun;Lee, JaeHee;Shin, SoJung;Cho, AhRang;Heo, Yong
    • Toxicological Research
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    • v.34 no.1
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    • pp.7-12
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    • 2018
  • Laboratory animal models have been developed to investigate preventive or therapeutic effect of medicinal products, or occurrence or progression mechanism of atopic dermatitis (AD), a pruritic and persistent inflammatory skin disease. The murine model with immunologic phenomena resembling human AD was introduced, which demonstrated skewedness toward predominance of type-2 helper T cell reactivity and pathophysiological changes similar as human AD following 2,4-dinitrochlorobenzene (DNCB) sensitization and challenge. Molecular mechanism on the DNCB-mediated AD was further evaluated. Skin tissues were collected from mice treated with DNCB, and each tissue was equally divided into two sections; one for protein and the other for mRNA analysis. Expression of filaggrin, an important protein for keratinocyte integrity, was evaluated through SDS-PAGE. Level of mRNA expression for cytokines was determined through semi-quantitative reverse transcriptase polymerase chain reaction. Expression of filaggrin protein was significantly enhanced in the mice treated with DNCB compared with the vehicle (acetone : olive oil = 4 : 1 mixture) treatment group or the normal group without any treatment. Level of tumor necrosis factor-alpha and interleukin-18 mRNA expression, cytokines involved in activity of type-1 helper T ($T_H1$) cell, was significantly downregulated in the AD group compared with other control groups. These results suggest that suppression of $T_H1$ cell-mediated immune response could be reflected into the skin tissue of mice treated with DNCB for AD induction, and disturbance of keratinocyte integrity might evoke a compensatory mechanism.

Identification of Genes that are Induced after Cadmium Exposure by Suppression Subtractive Hybridization

  • 이미옥
    • Proceedings of the Korea Society of Environmental Toocicology Conference
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    • 2003.05a
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    • pp.107-107
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    • 2003
  • The heavy metal cadmium is a xenobiotic toxicant of environmental and occupational concern and it has been classified as a human carcinogen. Inhalation of cadmium has been implicated in the development of emphysema and pulmonary fibrosis, but, the detailed mechanism by which cadmium induces adverse biological effects is not yet known. Therefore, we undertook the investigation of genes that are induced after cadmium exposure to illustrate the mechanism of cadmium toxicity For this purpose, we employed the polymerase chain reaction-based suppression subtractive hybridization technique. We identified 29 different cadmium-inducible genes in human peripheral mononuclear cells, such as macrophage migration inhibitory factor, lysophosphatidic acid acyltransferase-${\alpha}$, enolase-1${\alpha}$, VEGF, Bax, neuron-derived orphan receptor-1, and Nur77, which are known to be associated with inflammation, cell survival, and apoptosis. Induction of these genes by cadmium treatment was further confirmed by semi-quantitative reverse-transcription polymerase chain reaction. Further, we found that these genes were also induced after cadmium exposure in normal human lung fibroblast cell line, WI-38, suggesting potential use of this induction profile to monitor cadmium toxicity in the lung. Next, Nur77, one of cadmium-inducible genes, was further studied since the products of Nur77 are known to be involved in the apoptotic process of lung cells. Following cadmium treatment, Nur77 gene expression was increased at protein-level in A549 cells. Consistently, the reporter containing Nur77 binding sequence was activated by 2.5-fold after exposure to cadmium in reporter gene analysis by transient transfection experiments. When the plasmid encoding dominant negative Nur77 that represses the transcriptional function of wild-type Nur77 was transfected into A549 cells, the expression of Bax was significantly reduced, suggesting that induction of Nur77 was an important process in cadmium-induced apoptosis in the cells. Cadmium induced the expression of Nur77 in vivo, confirming the relevance of the data obtained in viro. Together our results suggest that Nur77 gene expression in exposure to cadmium leads apoptosis of lung cells which may cause pathological changes in lung.

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Effects of Genistein on the Gene Expressions of Glutathione Peroxidases and Superoxide Dismutases in Ethanol-Treated Mouse Fetuses

  • Yon, Jung-Min;Lin, Chunmei;Jung, A-Young;Lee, Jong-Geol;Jung, Ki-Youn;Baek, In-Jeoung;Lee, Beom-Jun;Nam, Sang-Yoon;Yun, Young-Won
    • Journal of Embryo Transfer
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    • v.26 no.2
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    • pp.135-140
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    • 2011
  • Genistein is a product of naturally occurring isoflavones at relatively high levels in soybeans. The harmful effects of ethanol are attributed to the induction of biological processes which lead to an increase in the generation of reactive oxygen species in fetuses. In this study, we investigated the effects of genistein ($1{\times}10^{-8}$ and $1{\times}10^{-7}\;{\mu}g$/ml) on gene expressions of the representative cellular antioxidative enzymes in ethanol (1 ${\mu}l$/ml)-treated mouse fetuses during the critical period (embryonic days 8.5~10.5) of organogenesis using a semi-quantitative RT-PCR analysis. The mRNA levels of cytosolic glutathione peroxidase (GPx), phospholipid hydroperoxide GPx, cytosolic CU,Zn-superoxide dismutase (SOD), and mitochondrial SOD were significantly decreased in ethanol-treated fetuses. However, the mRNA levels of ethanol plus genistein-treated fetuses were significantly higher than those of ethanol alone fetuses. These results indicate that genistein can up-regulate the expressions of GPx and SOD mRNAs reduced by the ethanol treatment in fetuses.

The Emotional Response and Coping Pattern by Age among Patients with Cancer (암 환자의 연령에 따른 정서반응과 대처양상)

  • Lee, Nae-Young;Kim, Young-Hae;Kim, Bock-Ryun;Kim, Jung-Sun
    • Asian Oncology Nursing
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    • v.5 no.1
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    • pp.22-30
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    • 2005
  • Purpose: The aim of this study was to investigate the differences in emotional response and coping pattern by age among cancer patients. Method: As descriptive research, from November 2000 to April 2001, data was collected with semi-structured questionnaire to 90 adult cancer patients, and analyzed using quantitative analysis. Result: Most emotional response at the time of diagnosis of cancer is despair in 20-39years & more than 60 years, and Impact in 40-59years. In emotional response during treatment by age, there were most much hope in 20-39 years, fear in 40-59years, and acceptance in more than 60years. In difficulties by age during treatment, there were most much mental burden in 20-29years, problems about occupation/finance in 40-59years, and physical discomfort related to treatment in more than 60 years. Resolution of difficulties of treatment shows avoidance in 20-39years, active participation in 40-59years and compliance in more than 60 years. Coping pattern during treatment was positive thinking in 20-39years, refreshment in 40-59years, and despair/avoidance in more than 60 years. Coping with treatment & progress shows in 20-39years maintenance of current health, 40-59years impossible to recover, more than 60year health recovery. Conclusion: Nursing could be considered emotional response and coping pattern according to age.

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Sonchus asper extract inhibits LPS-induced oxidative stress and pro-inflammatory cytokine production in RAW264.7 macrophages

  • Wang, Lan;Xu, Ming Lu;Liu, Jie;Wang, You;Hu, Jian He;Wang, Myeong-Hyeon
    • Nutrition Research and Practice
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    • v.9 no.6
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    • pp.579-585
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    • 2015
  • BACKGROUND/OBJECTIVES: Sonchus asper is used extensively as an herbal anti-inflammatory for treatment of bronchitis, asthma, wounds, burns, and cough; however, further investigation is needed in order to understand the underlying mechanism. To determine its mechanism of action, we examined the effects of an ethyl acetate fraction (EAF) of S. asper on nitric oxide (NO) production and prostaglandin-E2 levels in lipopolysaccharide (LPS)-stimulated RAW264.7 macrophages. MATERIALS/METHODS: An in vitro culture of RAW264.7 macrophages was treated with LPS to induce inflammation. RESULTS: Treatment with EAF resulted in significant suppression of oxidative stress in RAW264.7 macrophages as demonstrated by increased endogenous superoxide dismutase (SOD) activity and intracellular glutathione levels, decreased generation of reactive oxygen species and lipid peroxidation, and restoration of the mitochondrial membrane potential. To confirm its anti-inflammatory effects, analysis of expression of inducible NO synthase, cyclooxygenase-2, tumor necrosis factor-${\alpha}$, and the anti-inflammatory cytokines IL-$1{\beta}$ and IL-6 was performed using semi-quantitative RT-PCR. EAF treatment resulted in significantly reduced dose-dependent expression of all of these factors, and enhanced expression of the antioxidants MnSOD and heme oxygenase-1. In addition, HPLC fingerprint results suggest that rutin, caffeic acid, and quercetin may be the active ingredients in EAF. CONCLUSIONS: Taken together, findings of this study imply that the anti-inflammatory effect of EAF on LPS-stimulated RAW264.7 cells is mediated by suppression of oxidative stress.

Higher levels of serum triglyceride and dietary carbohydrate intake are associated with smaller LDL particle size in healthy Korean women

  • Kim, Oh-Yoen;Chung, Hye-Kyung;Shin, Min-Jeong
    • Nutrition Research and Practice
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    • v.6 no.2
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    • pp.120-125
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    • 2012
  • The aim of this study was to investigate the influencing factors that characterize low density lipoprotein (LDL) phenotype and the levels of LDL particle size in healthy Korean women. In 57 healthy Korean women (mean age, $57.4{\pm}13.1$ yrs), anthropometric and biochemical parameters such as lipid profiles and LDL particle size were measured. Dietary intake was estimated by a developed semi-quantitative food frequency questionnaire. The study subjects were divided into two groups: LDL phenotype A (mean size: $269.7{\AA}$, n = 44) and LDL phenotype B (mean size: $248.2{\AA}$, n = 13). Basic characteristics were not significantly different between the two groups. The phenotype B group had a higher body mass index, higher serum levels of triglyceride, total-cholesterol, LDL-cholesterol, apolipoprotein (apo)B, and apoCIII but lower levels of high density lipoprotein (HDL)-cholesterol and LDL particle size than those of the phenotype A group. LDL particle size was negatively correlated with serum levels of triglyceride (r = -0.732, $P$ < 0.001), total-cholesterol, apoB, and apoCIII, as well as carbohydrate intake (%En) and positively correlated with serum levels of HDL-cholesterol and ApoA1 and fat intake (%En). A stepwise multiple linear regression analysis revealed that carbohydrate intake (%En) and serum triglyceride levels were the primary factors influencing LDL particle size ($P$ < 0.001, $R^2$ = 0.577). This result confirmed that LDL particle size was closely correlated with circulating triglycerides and demonstrated that particle size is significantly associated with dietary carbohydrate in Korean women.