• 제목/요약/키워드: semi-quantitative analysis

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Artemisia capillaris Thunb. inhibits melanin synthesis activity via ERK-dependent MITF pathway in B16/F10 melanoma cells

  • Saba, Evelyn;Oh, Mi Ju;Lee, Yuan Yee;Kwak, Dongmi;Kim, Suk;Rhee, Man Hee
    • 대한수의학회지
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    • 제58권1호
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    • pp.1-7
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    • 2018
  • Genus Artemisia occurs as a hardy plant and has a wide range of culinary and medicinal features. In this study, we aimed to describe the melanin inhibitory activity of one Artemisia species, i.e., Artemisia capillaris Thunb. Ethanol extracts of fermented Artemisia capillaris (Art.EtOH.FT) and non-fermented Artemisia capillaris (Art.EtOH.CT) were tested for their ability to inhibit tyrosinase activity and melanin pigmentation. Both extracts showed dose-dependent inhibition against ${\alpha}$-melanocyte stimulating hormone-stimulated melanin formation and tyrosinase activity, without cytotoxicity. At $100{\mu}g/mL$, both extracts showed greater inhibition than kojic acid, the positive control. Protein expressions of microphthalmia-associated transcription factor (MITF), tyrosinase (TYR), tyrosinase-related protein 1 (TRP-1), and tyrosinase-related protein 2 (TRP-2) at the transcriptional level were determined by using real-time and semi-quantitative polymerase chain reaction. To complete the mechanistic study, presences of upstream elements of MITF, the phosphorylated-extracellular signal-regulated kinase (p-ERK), and phosphorylated-mitogen-activated protein kinase kinase (p-MEK) were confirmed by using western blot analysis. Expressions of p-TYR, p-TRP-1 and p-TRP-2, downstream factors for p-ERK and p-MITF, were translationally inhibited by both extracts. Art.EtOH.FT induced more potent effects than Art.EtOH.CT, especially signal transduction effects. In summary, Artemisia capillaris extracts appear to act as potent hypopigmentation agents.

50세 이상 중년 및 노인의 건강행위 요인에 따른 영양위험 연구 (Health-Related Behavioral Factors Associated with Nutritional Risks in Korean Aged 50 years and Over)

  • 임경숙
    • 대한지역사회영양학회지
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    • 제12권5호
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    • pp.592-605
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    • 2007
  • Undernutrition could be a significant deterrent to healthy aging and could negatively affect health outcomes in elderly. This study aimed to assess health-related factors which are associated with nutritional risks in middle-aged and elderly individuals by a cross-sectional study. Interviews were conducted with 2660 subjects (847 males, 1813 females), aged 50 years and over, in 15 cities in Korea. Data on food intake were obtained through a validated semi-quantitative food frequency questionnaires. Nutritional status were analyzed according to health-related factors including cigarette smoking, alcohol drinking, exercise, stress and depression level. Less regular exercise was associated with a higher likelihood of a poor nutrition [odds ratio (OR) 1.94; 95% confidence intervals (CI) 1.43-2.65] of middle-aged and elderly male subjects. Cigarette smoking (OR 1.84; 95% CI 1.24-2.71), less exercise (OR 2.58; 95% CI 2.07-3.21), stress (OR 1.73; 95% CI 1.36-2.22), and depression (OR 1.34, 95% CI 1.08-1.67) of middle-aged and elderly female subjects was associated with a higher likelihood of a poor nutrition. The results of the multiple regression analysis showed that less exercise proved to be the strongest predictors for the poor nutrition, followed by stress, smoking, and depression (model $R^2= 9.0%$). It suggests that guidance to promote regular exercise, to quit smoking, to minimize stress and depression level might help to improve nutritional status of middle-aged and elderly in Korea. These findings also suggest that having recommendable health behaviors are beneficial to the good nutrition of subjects aged 50 years and over.

Cloning and Characterization of ${\Delta}^1$-Pyrroline-5-Carboxylate Synthetase Genes and Identification of Point Mutants in Medicago truncatula

  • Song, Ki-Hoon;Song, Dae-Hae;Lee, Jeong-Ran;Kim, Goon-Bo;Choi, Hong-Kyu;Penmetsa, R. Varma;Nam, Young-Woo
    • 한국작물학회지
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    • 제52권4호
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    • pp.458-468
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    • 2007
  • To tolerate environmentally adverse conditions such as cold, drought, and salinity, plants often synthesize and accumulate proline in cells as compatible osmolytes. ${\Delta}^1$-Pyrroline-5-carboxylate synthetase(P5CS) catalyzes the rate-limiting step of proline biosynthesis from glutamate. Two complete genes, MtP5CS1 and MtP5CS2, were isolated from the model legume Medicago truncatula by cDNA cloning and bacterial artificial chromosome library screening. Nucleotide sequence analysis showed that both genes consisted of 20 exons and 19 introns. Alignment of the predicted amino acid sequences revealed high similarities with P5CS proteins from other plant species. The two MtP5CS genes were expressed in response to high salt and low temperature treatments. Semi-quantitative reverse transcription-polymerase chain reaction showed that MtP5CS1 was expressed earlier than MtP5CS2, indicating differential regulation of the two genes. To evaluate the reverse genetic effects of nucleotide changes on MtP5CS function, a Targeting Induced Local Lesions in Genomes approach was taken. Three mutants each were isolated for MtP5CS1 and MtP5CS2, of which a P5CS2 nonsense mutant carrying a codon change from arginine to stop was expected to bring translation to premature termination. These provide a valuable genetic resource with which to determine the function of the P5CS genes in environmental stress responses of legume crops.

Oily Fish Consumption Modifies the Association between CD36 rs6969989 Polymorphism and Lipid Profiles in Korean Women

  • Shin, Yoonjin;Kim, Yangha
    • Preventive Nutrition and Food Science
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    • 제21권3호
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    • pp.202-207
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    • 2016
  • The aim of this study was to investigate the association of CD36, a class B scavenger receptor, rs6969989 polymorphism with the serum lipid profiles in Korean women, together with their modulation by oily fish consumption. Subjects were participants from the Korean Genome Epidemiology Study (KoGES), which was initiated in 2001 as a large-scale. A total of 4,210 women aged 39 to 70 were included in this study. Data were collected using self-administered questionnaires, anthropometric measurements, and blood chemical analysis. Dietary intake was analyzed using a semi-quantitative food frequency questionnaire. The minor allele frequency for rs6969989 was found in 12% of this population. Homozygotes minor G allele at the rs6868989 exhibited significantly higher high density lipoprotein cholesterol (HDLC) concentrations (P-trend=0.043) and lower fasting glucose (P-trend=0.013) than major allele A carriers. The risk of low HDL-C was significantly lower in homozygotes for the G allele than the A allele carriers (P-trend=0.032). Gene-diet interaction effects between rs6969989 and oily fish intake were significantly associated with the risk of dyslipidemia (P-interaction=0.004). Subjects with homozygotes minor G allele and high oily fish intake generally had a lower risk of dyslipidemia than did those with major allele homozygotes and low oily fish intake. These findings supported that oily fish consumption may modulate the contributions of CD36 rs6969989 on genetic predisposition to the risk of dyslipidemia.

주요 간호활동의 건강보험 수가 적절성에 관한 연구 (A Study on the Appropriateness of Health Insurance Fee in Main Nursing Practices)

  • 김윤미;김경숙;박미미;김인숙;김미영
    • 임상간호연구
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    • 제23권2호
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    • pp.236-247
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    • 2017
  • Purpose: This study was to present improvement strategy and the problems of the nursing fee in national health insurance system. Methods: A total of 23 nursing activities performed by nurses were selected. Data were collected the relative value score and criteria of the Health Insurance Review & Assessment Agency. Sixty clinical nursing experts panels were composed and nursing time surveyed self-reported method. The actual fee was calculated through the nursing time, relative value score and actual labor costs. Nextly, the labor costs analyzed was compared with that in the national health insurance. Results: Although the practices were mainly performed by the nurse, other occupations have been recorded as main practitioners and the time of the nursing activity is partially improperly reflected. Additionally, although the nurse practiced mainly in glucose (semi-quantitative) test, it was confirmed that the principal practitioner was described as a clinical pathologist. The the labor cost gap was estimated that is 9.3 times (median) and 11.9 times (average) in this analysis. Conclusion: This study suggests that it is necessary to legislate a policy that can improve the quality of clinical nursing by reinforcing the appropriateness and improving nursing fee through reflection of the actual time spent for nursing care.

Screening of Domestic Silkworm Strains for Efficient Heterologous Protein Expression by Bombyx mori Nuclear Polyhedrosis Virus (BmNPV)

  • Jo, Sun Jung;Choi, Ji-Hyun;Kang, Ju-Il;Lim, Jae-Hwan;Seok, Young Sik;Lee, Jae Man;Kusakabe, Takahiro;Hong, Sun Mee
    • International Journal of Industrial Entomology and Biomaterials
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    • 제29권2호
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    • pp.185-192
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    • 2014
  • Recombinant proteins can be generated quickly and easily in large amounts and at low-cost in silkworm larvae by using Bombyx mori nuclear polyhedrosis virus (BmNPV). We searched for high-permissive silkworm strains that have high production levels of heterologous proteins and are thus suitable for use as biofactories. In this study, we performed the analysis using a BmNPV vector expressing luciferase as a marker, and we confirmed protein expression by evaluating luciferase activity, determined by western blotting and luciferase ELISA, and confirmed transcription expression by semi- and quantitative real time PCR. For the selection of host silkworm strains, we first chose 52 domestic BmNPV sensitive strains and then identified 10 high-permissive and 5 low-permissive strains. In addition, to determine which hybrid of the high-permissive strains would show heterosis, nine strains derived through three-way crossing were tested for luciferase activity by western blotting, and luciferase ELISA. We found a correlation between luciferase activity and luciferase protein expression, but not transcription. There was no noticeable difference in protein expression levels between Jam313 as the high-permissive control strain and the three-way hybrid strains; however, the three-way cross strains showed lower luciferase activity compared with Jam313. In this study, luciferase protein production in the larvae of 52 domestic silkworm strains was elucidated using BmNPV.

Transcriptional Regulation of the Methuselah Gene by Dorsal Protein in Drosophila melanogaster

  • Kim, Hyukmin;Kim, Jinsu;Lee, Yoonsoo;Yang, Jaeyeon;Han, Kyuhyung
    • Molecules and Cells
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    • 제21권2호
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    • pp.261-268
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    • 2006
  • The Drosophila methuselah (mth) mutant has an approximately 35 percent increase in average lifespan, and enhanced resistance to various forms of stress, including starvation, high temperature, and dietary paraquat. To examine the transcriptional regulation of mth, we used luciferase assays employing Drosophila S2 cells. Two positive control elements were found at -542 ~ -272 (PE1) and +28 ~ +217 (PE2), where putative binding sites for transcription factors including Dorsal (Dl) were identified. Cotransfection of a Dl expression plasmid with a mth-luciferase reporter plasmid resulted in decreased reporter activity. PE1 and PE2, the minimal elements for strong promoter activity, were required for maximal repression by Dl protein. The N-terminal Rel homology domain (RHD) of Dl was not sufficient for repression of mth. We demonstrated by chromatin affinity precipitation (ChAP) assays in S2 cells that Dl bound to the putative PE1 binding site. Unexpectedly, semi-quantitative RT-PCR analysis revealed that the level of mth transcripts was reduced in dl flies. However, the in vivo result support the view that mth expression is regulated by dl, since it is well known that Dl functions as both a transcriptional activator and repressor depending on what other transcription factors are present. These findings suggest that both innate immunity and resistance to stress are controlled by Dl protein.

성인 여성을 대상으로 한 지방섭취제한행동 변화단계에 따른 사회심리적 요인 분석 (Analysis on the Stages of Change in Fat Reducing Behavior and Social Psychological Correlates in adult Female)

  • 오세영;조미란;김진옥
    • 대한지역사회영양학회지
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    • 제5권4호
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    • pp.615-623
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    • 2000
  • Under the assumption that people go through stages in making dietary behavior change, this study was attempted to apply the stages of a change model for fat intake by examining the associations of social psychological variables with stages of change in dietary fat reduction. Derived from social psychological theories, 10 social psychological variables on motivational beliefs(6), social influence(3) and self-efficacy(1) related specifically to selecting every day diets low in fat were constructed. Fat and energy intakes were assessed by a short form semi-quantitative food frequency questionnaire. The associations of stages of change with motivational beliefs, social influence, and self-efficacy variables and energy and fat intakes were assessed in 333 female adults from large cities in Korea. Dietary stage groups differed significantly on most of the social psychological variables in ways predicted by theory. Motivational factors that lead to a psychological state of readiness to take action were important in the early stages of the dietary change process. Social influences were more important in the stages as people decide to take action. Self efficacy and motivational beliefs, particularly, reduction of perceived barriers were important in maintaining fat reduction behavior. The results of our study indicate differences in stages of change in fat reduction behavior in terms of nutrient intakes and social psychological correlates and suggests that adding a time dimension to social psychological models increases our understanding of dietary change, which assist us in designing nutrition education interventions that are more appropriately targeted by stage of change. (Korean J Community Nutrition 5(4) 615∼623, 2000)

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Expression of a Glutathione Reductase from Brassica rapa subsp. pekinensis Enhanced Cellular Redox Homeostasis by Modulating Antioxidant Proteins in Escherichia coli

  • Kim, Il-Sup;Shin, Sun-Young;Kim, Young-Saeng;Kim, Hyun-Young;Yoon, Ho-Sung
    • Molecules and Cells
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    • 제28권5호
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    • pp.479-487
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    • 2009
  • Glutathione reductase (GR) is an enzyme that recycles a key cellular antioxidant molecule glutathione (GSH) from its oxidized form (GSSG) thus maintaining cellular redox homeostasis. A recombinant plasmid to overexpress a GR of Brassica rapa subsp. pekinensis (BrGR) in E. coli BL21 (DE3) was constructed using an expression vector pKM260. Expression of the introduced gene was confirmed by semi-quantitative RT-PCR, immunoblotting and enzyme assays. Purification of the BrGR protein was performed by IMAC method and indicated that the BrGR was a dimmer. The BrGR required NADPH as a cofactor and specific activity was approximately 458 U. The BrGR-expressing E. coli cells showed increased GR activity and tolerance to $H_2O_2$, menadione, and heavy metal ($CdCl_2$, $ZnCl_2$ and $AlCl_2$)-mediated growth inhibition. The ectopic expression of BrGR provoked the co-regulation of a variety of antioxidant enzymes including catalase, superoxide dismutase, glutathione peroxidase, and glucose-6-phosphate dehydrogenase. Consequently, the transformed cells showed decreased hydroperoxide levels when exposed to stressful conditions. A proteomic analysis demonstrated the higher level of induction of proteins involved in glycolysis, detoxification/oxidative stress response, protein folding, transport/binding proteins, cell envelope/porins, and protein translation and modification when exposed to $H_2O_2$ stress. Taken together, these results indicate that the plant GR protein is functional in a cooperative way in the E. coli system to protect cells against oxidative stress.

흡연과 Carotenoid 함유 식품 섭취빈도에 따른 지질과산화 및 항산화능 비교 (Effect of Smoking and Carotenoid-rich Food Consumption on Lipid Peroxidation and Antioxidant Status)

  • 김유경;천종희
    • Journal of Nutrition and Health
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    • 제38권10호
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    • pp.836-846
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    • 2005
  • This study was performed to determine the effect of carotenoid-rich food consumption and smoking on the lipid peroxidation and antioxidant status in human. The subjects consisted of the health 210 middle-aged adults who visited health care center in the Inha University hospital. The blood and urine samples of the subjects were taken to analyze serum lipid profiles, plasma TBARS, total antioxidant status (TAS) and urinary 8-isoprostanes concentration. The anthropometric indices of the subjects were measured. The nutritional intake and the frequency of carotenoid-rich food consumption was determined by semi-quantitative food frequency questionnaire survey. HDL-cholesterol level of the smoking subjects was significantly lower than that of the non-smoking subjects in men. In the analysis of the carotenoid-rich food consumption, the frequency of pepper and tomato consumption of the non-smoking subjects was significantly higher than that of the smoking subjects in men. In women, the Sequency of pear and peach consumption of the non-smoking subjects was significantly higher than that of the smoking subjects. HDL-cholesterol level of the high carotenoid rich food consumption group was significantly higher than that of medium and low group in women. TAS of the high carotenoid rich food consumption group was significantly higher than that of medium and low group. In conclusion, carotenoid intakes seemed to be effective to reduce lipid peroxidation and increase antioxidant status in the body. The frequency of the carotenoid-rich food consumption seemed to be lower in the smokers than in the non-smokers. However, further researches consisted of large-scaled and randomized clinical trials are required to determine whether carotenoids have any other beneficial effect in human. (Korean J Nutrition 38(10): 836$\sim$846,2005)