• 제목/요약/키워드: calcium sensitivity

검색결과 82건 처리시간 0.025초

Establishment of a NanoBiT-Based Cytosolic Ca2+ Sensor by Optimizing Calmodulin-Binding Motif and Protein Expression Levels

  • Nguyen, Lan Phuong;Nguyen, Huong Thi;Yong, Hyo Jeong;Reyes-Alcaraz, Arfaxad;Lee, Yoo-Na;Park, Hee-Kyung;Na, Yun Hee;Lee, Cheol Soon;Ham, Byung-Joo;Seong, Jae Young;Hwang, Jong-Ik
    • Molecules and Cells
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    • 제43권11호
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    • pp.909-920
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    • 2020
  • Cytosolic Ca2+ levels ([Ca2+]c) change dynamically in response to inducers, repressors, and physiological conditions, and aberrant [Ca2+]c concentration regulation is associated with cancer, heart failure, and diabetes. Therefore, [Ca2+]c is considered as a good indicator of physiological and pathological cellular responses, and is a crucial biomarker for drug discovery. A genetically encoded calcium indicator (GECI) was recently developed to measure [Ca2+]c in single cells and animal models. GECI have some advantages over chemically synthesized indicators, although they also have some drawbacks such as poor signal-to-noise ratio (SNR), low positive signal, delayed response, artifactual responses due to protein overexpression, and expensive detection equipment. Here, we developed an indicator based on interactions between Ca2+-loaded calmodulin and target proteins, and generated an innovative GECI sensor using split nano-luciferase (Nluc) fragments to detect changes in [Ca2+]c. Stimulation-dependent luciferase activities were optimized by combining large and small subunits of Nluc binary technology (NanoBiT, LgBiT:SmBiT) fusion proteins and regulating the receptor expression levels. We constructed the binary [Ca2+]c sensors using a multicistronic expression system in a single vector linked via the internal ribosome entry site (IRES), and examined the detection efficiencies. Promoter optimization studies indicated that promoter-dependent protein expression levels were crucial to optimize SNR and sensitivity. This novel [Ca2+]c assay has high SNR and sensitivity, is easy to use, suitable for high-throughput assays, and may be useful to detect [Ca2+]c in single cells and animal models.

한국 성인 남녀 대사증후군 집단의 영양소 섭취와 식사의 질 및 hs-CRP와 관련성 - 국민건강영양조사(2015년) 자료를 활용하여 - (Relationship between Nutrients Intakes, Dietary Quality, and hs-CRP in Korea Metabolic Syndrome Patients - The 2015 Korea National Health and Nutrition Examination Survey -)

  • 김미성;김병숙;이종신;오경재;한성희
    • 한국식품영양학회지
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    • 제31권3호
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    • pp.425-434
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    • 2018
  • Metabolic syndrome is a risk factor for cardiovascular and type 2 diabetes. This study was conducted to examine the relevance between nutrition intake, meal quality, and high-sensitivity C-reactive protein in Koreans with metabolic syndrome. The 2,536 subjects, aged 19~64, who participated in 2015 National Nutrition Survey were included in this study. The 24-hour recall method was employed to analyze nutrition intake and dietary quality. Subjects were grouped into either the non-metabolic syndrome group (n=1,938) or the metabolic syndrome group (n=598). Total males and females were divided into 3 groups according to the high-sensitivity C-reactive protein (hs-CRP) level to study its relationship to metabolic syndrome and its components, including odds ratio (OR) and confidence interval (CI). Results showed the homeostasis model assessment of insulin resistance (HOMA-IR) value was higher in the metabolic syndrome group (3.37) than non-metabolic syndrome group (1.57) (p<0.001). In the Index of Nutrition Quality, males in the non-metabolic syndrome group showed higher niacin (p<0.05) than males in metabolic syndrome group. Females in the non-metabolic syndrome group had higher vitamin $B_1$ (p<0.01), vitamin $B_2$ (p<0.001), niacin (p<0.05), calcium (p<0.001), and phosphate (p<0.01). Female in the high hs-CRP group showed high OR in blood glucose component (OR 2.488, 95% CI: 1.269~4.879) and metabolic syndrome risk (OR 2.856, 95% CI: 1.292~6.314). Females in the middle hs-CRP group had high triglycerides component (OR 2.956, 95% CI: 1.920~4.551), compared to the low hs-CRP group. The study showed females with higher hs-CRP had a higher risk of metabolic syndrome.

한국 성인의 식사 패턴과 인슐린 저항성 간의 상관성: 2015년도 국민건강영양조사를 이용하여 (The association of dietary patterns with insulin resistance in Korean adults: based on the 2015 Korea National Health and Nutrition Examination Survey)

  • 김이슬;양윤정
    • Journal of Nutrition and Health
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    • 제54권3호
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    • pp.247-261
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    • 2021
  • 본 연구는 제6기 2015년도 국민건강영양조사 자료를 활용하여 만 19-64세 한국 성인의 주요 식사 패턴을 도출한 후, 식사 패턴과 인슐린 저항성 간의 상관성을 분석하기 위해 수행되었다. 그 결과는 다음과 같다. 대상자의 식사 패턴을 요인분석 방법으로 분석한 결과, 'Healthy Korean meal pattern', 'Western meal pattern', 'White rice, alcohol, meat pattern'으로 나타났다. 'Healthy Korean meal pattern' 점수가 높을수록 연령이 높게 나타났으며, 식사 패턴에 따른 영양소 섭취량 분석 결과, 'Healthy Korean meal pattern' 섭취 점수가 높을수록 총 에너지, 탄수화물, 단백질, 비타민 A, 비타민 C, 칼슘, 인, 철, 칼륨, 식이섬유의 섭취가 유의하게 증가하였으며 'Western meal pattern' 섭취 점수가 높을수록 총 에너지, 단백질, 지방, 콜레스테롤의 섭취가 유의하게 증가하였고, 'White rice, alcohol, meat pattern' 점수가 높을수록 총 에너지와 나트륨의 섭취는 유의하게 증가하였고, 탄수화물, 비타민 C, 칼슘, 인, 칼륨, 식이섬유의 섭취는 유의하게 감소하였다. 식사 패턴에 따른 인슐린 저항성의 위험도를 분석한 결과 'Healthy Korean meal pattern'과 'Western meal pattern'은 인슐린 저항성과 유의한 연관성을 보이지 않았고, 'White rice, alcohol, meat pattern'은 인슐린 저항성의 위험도와 양의 연관성을 보였다. 이상의 결과로부터 'White rice, alcohol, meat pattern'의 식사를 많이 할수록 인슐린 저항성의 위험도가 높아지는 것을 알 수 있었다. 본 연구는 단면연구라는 제한점이 있으므로 향후 식사 패턴과 인슐린 저항성의 상관성을 확인할 수 있는 전향적 코호트 등의 연구가 필요하다.

Enhancement of β-cyclodextrin Production and Fabrication of Edible Antimicrobial Films Incorporated with Clove Essential Oil/β-cyclodextrin Inclusion Complex

  • Farahat, Mohamed G.
    • 한국미생물·생명공학회지
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    • 제48권1호
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    • pp.12-23
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    • 2020
  • Edible films containing antimicrobial agents can be used as safe alternatives to preserve food products. Essential oils are well-recognized antimicrobials. However, their low water solubility, volatility and high sensitivity to oxygen and light limit their application in food preservation. These limitations could be overcome by embedding these essential oils in complexed product matrices exploiting the encapsulation efficiency of β-cyclodextrin. This study focused on the maximization of β-cyclodextrin production using cyclodextrin glucanotransferase (CGTase) and the evaluation of its encapsulation efficacy to fabricate edible antimicrobial films. Response surface methodology (RSM) was used to optimize CGTase production by Brevibacillus brevis AMI-2 isolated from mangrove sediments. This enzyme was partially purified using a starch adsorption method and entrapped in calcium alginate. Cyclodextrin produced by the immobilized enzyme was then confirmed using high performance thin layer chromatography, and its encapsulation efficiency was investigated. The clove oil/β-cyclodextrin inclusion complexes were prepared using the coprecipitation method, and incorporated into chitosan films, and subjected to antimicrobial testing. Results revealed that β-cyclodextrin was produced as a major product of the enzymatic reaction. In addition, the incorporation of clove oil/β-cyclodextrin inclusion complexes significantly increased the antimicrobial activity of chitosan films against Staphylococcus aureus, Staphylococcus epidermidis, Salmonella Typhimurium, Escherichia coli, and Candida albicans. In conclusion, B. brevis AMI-2 is a promising source for CGTase to synthesize β-cyclodextrin with considerable encapsulation efficiency. Further, the obtained results suggest that chitosan films containing clove oils encapsulated in β-cyclodextrin could serve as edible antimicrobial food-packaging materials to combat microbial contamination.

증감지(增感紙)-필름계(系)의 상대감도(相對感度)와 절대감도(絶對感度)에 관한 실험(實驗) (The Relative and Absolute Speed of Radiographic Screen-Film Systems)

  • 허준;이인자
    • 대한방사선기술학회지:방사선기술과학
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    • 제16권1호
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    • pp.67-80
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    • 1993
  • Recently, a large number of new screen-film systems have become available for use in diagnostic radiology. These new screens are made of materials generally known as rare-earth phosphors which have high x-ray absorption and high x-ray to light conversion efficiency compared to calcium tungstate phosphors. The major advantage of these new systesms is reduction of patient exposure due to their high speed or high sensitivity. However, a system with excessively high speed can result in a significant degradation of radiographic image quality. Therefore, th speed is important parameters for users of these system. Our aim of in this was to determine accurately and precisely the absolute speed and relative speeds of both new and conventional screen-film system. We determined the absolute speed in condition of BRH phantom beam qulity and the relative speed were measured by a split-screen technique in condition of BRH and ANSI phantom beam qulity. The absolute and the relative speed were determined for 8 kinds of screen-4 kinds of film in regular system and 7 kinds of screen-7 kinds of film in ortho system. In this study we could know the New Rx, T-MAT G has the highest film speed, also know Green system's stndard deviation of relative speed larger than blue system. It was realized that there were no relationship between the absolute speed and the blue system. It was realized that there were no relationship between the absolute speed and the relative speed in ortho or regular system.

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A Gene Encoding Phosphatidyl Inositol-specific Phospholipase C form Cryphonectria parasitica Modulates the Hypoviral-modulated Laccase1 Expression

  • Kim, Dae-Hyuk
    • 한국미생물학회:학술대회논문집
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    • 한국미생물학회 2005년도 International Meeting of the Microbiological Society of Korea
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    • pp.159-161
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    • 2005
  • Hypovirus infection of the chestnut blight fungus Cryphonectria parasitica is a useful model system to study the hypoviral regulation of fungal gene expression. The hypovirus is known to downregulate the fungal laccase1 (lac 1), the modulation of which is tightly governed by the inositol triphosphate ($IP_3$) and calcium second messenger system in a virus-free strain. We cloned the gene cplc1 encoding a phosphatidyl inositol-specific phospholipase C (PLC), in order to better characterize the fungal gene regulation by hypovirus. Sequence analysis of the cplc1 gene indicated that the protein product contained both the X and Y domains, which are the two conserved regions found in all known PLCs, with a 133 amino acid extension between the 2nd ${\beta}$-strand and the ${\alpha}$-helix in the X domain. In addition, the gene organization appeared to be highly similar to that of a ${\delta}$ type PLC. Disruption of the cplc1 gene resulted in slow growth and produced colonies characterized by little aerial mycelia and deep orange in color. In addition, down regulation of lac1 expression was observed. However, temperature sensitivity, osmosensitivity, virulence, and other hypovirulence-associated characteristics did not differ from the wild-type strain. Functional complementation of the cplc1-null mutant with the PLC1 gene from Saccharomyces cerevisiae restored lac1 expression, which suggests that the cloned gene encodes PLC activity. The present study indicates that the cplc1 gene is required for appropriate mycelial growth, and that it regulates the lac1 expression, which is also modulated by the hypovirus. Although several PLC genes have been identified in various simple eukaryotic organisms, the deletion analysis of the cplc1 gene in this study appears to be the first report on the functional analysis of PLC in filamentous fungi.

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Application of artificial neural networks to predict total dissolved solids in the river Zayanderud, Iran

  • Gholamreza, Asadollahfardi;Afshin, Meshkat-Dini;Shiva, Homayoun Aria;Nasrin, Roohani
    • Environmental Engineering Research
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    • 제21권4호
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    • pp.333-340
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    • 2016
  • An Artificial Neural Network including a Radial Basis Function (RBF) and a Time Delay Neural Network (TDNN) was used to predict total dissolved solid (TDS) in the river Zayanderud. Water quality parameters in the river for ten years, 2001-2010, were prepared from data monitored by the Isfahan Regional Water Authority. A factor analysis was applied to select the inputs of water quality parameters, which obtained total hardness, bicarbonate, chloride and calcium. Input data to the neural networks were pH, $Na^+$, $Mg^{2+}$, Carbonate ($CO{_3}^{-2}$), $HCO{_3}^{-1}$, $Cl^-$, $Ca^{2+}$ and Total hardness. For learning process 5-fold cross validation were applied. In the best situation, the TDNN contained 2 hidden layers of 15 neurons in each of the layers and the RBF had one hidden layer with 100 neurons. The Mean Squared Error and the Mean Bias Error for the TDNN during the training process were 0.0006 and 0.0603 and for the RBF neural network the mentioned errors were 0.0001 and 0.0006, respectively. In the RBF, the coefficient of determination ($R^2$) and the index of agreement (IA) between the observed data and predicted data were 0.997 and 0.999, respectively. In the TDNN, the $R^2$ and the IA between the actual and predicted data were 0.957 and 0.985, respectively. The results of sensitivity illustrated that $Ca^{2+}$ and $SO{_4}^{2-}$ parameters had the highest effect on the TDS prediction.

Comparative evaluation of NovaMin desensitizer and Gluma desensitizer on dentinal tubule occlusion: a scanning electron microscopic study

  • Joshi, Surabhi;Gowda, Ashwini Shivananje;Joshi, Chintan
    • Journal of Periodontal and Implant Science
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    • 제43권6호
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    • pp.269-275
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    • 2013
  • Purpose: In this study, the effect of calcium sodium phosphosilicate (NovaMin) desensitizing agent, which is a powder-based system, and hydroxyethyl methacrylate and glutaraldehyde (Gluma desensitizer), which is liquid-based system, on dentinal tubule occlusion was analyzed by scanning electron microscope. The effects of the above two along with one control group were compared to determine the more effective method of sealing the dentinal tubules after initial application. Methods: Twenty specimens were allocated to each of 3 groups: Control, Gluma desensitizer, and NovaMin. Two additional samples were also prepared and treated with Gluma and NovaMin; these samples were longitudinally fractured. The specimens were prepared from extracted sound human premolars and were stored in 10% formalin at room temperature. The teeth were cleaned of gross debris and then sectioned to provide one to two dentin specimens. The dentin specimens were etched with 6% citric acid for 2 minutes and rinsed in distilled water. Control discs were dried, and the test discs were treated with the desensitizing agents as per the manufacturer's instructions. The discs as well as longitudinal sections were later analyzed under the scanning electron microscope. The proportions of completely occluded, partially occluded, and open tubules within each group were calculated. The ratios of completely and partially occluded tubules to the total tubules for all the groups was determined, and the data was statistically analyzed using nonparametric tests and statistical significance was calculated. Results: NovaMin showed more completely occluded tubules ($0.545{\pm}0.051$) while Gluma desensitizer showed more partially occluded tubules ($0.532{\pm}0.075$). The differences among all the groups were statistically significant ($P{\leq}0.05$). Conclusion: Both materials were effective in occluding dentinal tubules but NovaMin appeared more promising in occluding tubules completely after initial application.

토양중(土壤中) 2,4-D와 MCPP의 잔류특성(殘留特性) (Persistence of 2,4-D and MCPP in Soil)

  • 이영득;이해근;박영선
    • 한국환경농학회지
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    • 제2권2호
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    • pp.59-64
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    • 1983
  • 수원소재(水原所在) 2개토양(個土壤)에서 2,4-D ethyl ester 및 MCPP의 토양중(土壤中) 잔유소장(殘油消長)을 조사(調査)한 결과(結果)는 다음과 같다. 1) 살포(撒布)한 2,4-D ethyl ester는 토양중(土壤中)에서 신속(迅速)히 가수분해(加水分解)되어 2,4-D를 생성(生成)하였다. 2) 포장조건하(圃場條件下)에서 2,4-D 잔유분(殘留分)을 포함한 총(總) 2,4-D ethyl ester의 반감기(半減期)는 $4{\sim}7$일(日)이었으며 실험실(實驗室) 조건하(條件下)에서는 다소 긴 $4{\sim}7$일(日)이었다. 3) 포장조건하(圃場條件下)에서의 토양중(土壤中) MCPP의 반감기(半減期)는 $9{\sim}12$일(日)이었고 실험실(實驗室) 조건하(條件下)에서는 다소 긴 $12{\sim}23$일(日)의 반감기(半減期)를 보였다.

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L6 근육세포에서 포도당 수송능에 미치는 $CdCl_2$의 영향 (Effects of Cadmium on Glucose Transport in L6 Myocytes)

  • 강동희;길이룡;박광식;이병훈;문창규
    • Environmental Analysis Health and Toxicology
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    • 제20권1호
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    • pp.75-85
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    • 2005
  • This study was aimed to know the effect of cadmium chloride (CdCl₂) on glucose transport in L6 myotube and its action mechanism. CdCl₂ increased the 2-deoxy- (l-3H)-D-glucose (2-DOG) uptake 1.9 and 2.4 fold at 10 and 25 μM respectively. To investigate the stimulating-mechanism of glucose transport induced by CdCl₂, the wortmannin and PD98059 were used as PI3K (phosphatidylinositol 3-kinase) inhibitor and MAPK inhibitor respectively, which did not affect 2-DOG uptake. This fact suggests that CdCl₂ induced 2-DOG uptake may not be concerned to the insulin signalling pathway. Whereas nifedipine, a calcium channel blocker, and trifluoperazine, a calmodulin inhibitor, were found to inhibit the 2-DOG uptake stimulted by CdCl₂. In addition, we also measured the ROS (reactive oxygen species) production and GSH level in L6 myotube to investigate the correlation between the glucose uptake and ROS. CdCl₂(25 μM) increased ROS generation approximately 1.5 fold and changed the cellular GSH level, but GSSG/GSH ratio remained unchanged. CdCl₂ stimulated 2-DOG uptake and ROS generation were inhibited by N-acetylcystein. And BSO pretreatment, a potent inhibitor of γ-GCS, resulted in the dramatic decrease of 2-DOG uptake and also the increase of the sensitivity to cadmium cytotoxicity. The obtained results suggest that CdCl₂-stimulated glucose uptake might be based on the activation of HMP shunt as an antioxidant defense mechanism of the cells.