• 제목/요약/키워드: Metabolic Analysis

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Energy Metabolism in Human Pluripotent Stem and Differentiated Cells Compared Using a Seahorse XF96 Extracellular Flux Analyzer

  • Hyun Kyu Kim;Yena Song;Minji Kye;Byeongho Yu;Sang Beom Park;Ji Hyeon Kim;Sung-Hwan Moon;Hyungkyu Choi;Jong-Seok Moon;Jae Sang Oh;Man Ryul Lee
    • International Journal of Stem Cells
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    • 제17권2호
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    • pp.194-203
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    • 2024
  • Evaluating cell metabolism is crucial during pluripotent stem cell (PSC) differentiation and somatic cell reprogramming as it affects cell fate. As cultured stem cells are heterogeneous, a comparative analysis of relative metabolism using existing metabolic analysis methods is difficult, resulting in inaccuracies. In this study, we measured human PSC basal metabolic levels using a Seahorse analyzer. We used fibroblasts, human induced PSCs, and human embryonic stem cells to monitor changes in basal metabolic levels according to cell number and determine the number of cells suitable for analysis. We evaluated normalization methods using glucose and selected the most suitable for the metabolic analysis of heterogeneous PSCs during the reprogramming stage. The response of fibroblasts to glucose increased with starvation time, with oxygen consumption rate and extracellular acidification rate responding most effectively to glucose 4 hours after starvation and declining after 5 hours of starvation. Fibroblasts and PSCs achieved appropriate responses to glucose without damaging their metabolism 2~4 and 2~3 hours after starvation, respectively. We developed a novel method for comparing basal metabolic rates of fibroblasts and PSCs, focusing on quantitative analysis of glycolysis and oxidative phosphorylation using glucose without enzyme inhibitors. This protocol enables efficient comparison of energy metabolism among cell types, including undifferentiated PSCs, differentiated cells, and cells undergoing cellular reprogramming, and addresses critical issues, such as differences in basal metabolic levels and sensitivity to normalization, providing valuable insights into cellular energetics.

Clinical Usefulness of Serum Uric Acid and Resting Heart Rate in the Diagnosis of Metabolic Syndrome in Korean Adults

  • Shin, Kyung-A
    • 대한의생명과학회지
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    • 제23권2호
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    • pp.118-127
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    • 2017
  • Elevated serum uric acid and resting heart rate are risk factors and predictors of metabolic syndrome. However, few studies have examined the optimal cutoff value for serum uric acid and resting heart rate to predict metabolic syndrome in Korean adults. Subjects for this study were 22,302 adults (average age 45 years old), who underwent health screening examination from January 2010 to December 2012 at the Health Promotion Center of one hospital in Gyeonggi-do for general health check-up. The uric acid and resting heart rate cutoff values were calculated by ROC analysis for metabolic syndrome. Elevated serum uric acid and resting heart rate were associated with an increased prevalence of metabolic syndrome in Korean adults. The optimal cutoff value for uric acid level to predict metabolic syndrome in adults was 4.95 mg/dL (male 6.35, female 4.55) and optimal cutoff value for resting heart rate to predict metabolic syndrome was 68 beats per minute (male 66, female 68). However, serum uric acid and resting heart rate were found to have limitations for the diagnosis of metabolic syndrome.

일부지역 근로자의 대사증후군 위험요인별 분류와 구강건강상태 비교 (Comparison of oral health status by metabolic syndrome risk factors in workers)

  • 구인영;문선정;이명선;가경환
    • 한국치위생학회지
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    • 제13권4호
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    • pp.581-588
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    • 2013
  • Objectives : The aim of the study si to investigate correlation between metabolic syndrome risk factors and oral health status in workers and to propose the integrated and effective management measures. Methods : Subjects were 4,600 workers in industrial company. Data by medical checkup and oral examination were collected from July 13, 2010 to September 12, 2010. Self-reported questionnaire included general characteristics, job description, smoking, drinking, and metabolic syndrome risk factor indicators. Gingivitis, dental calculus and missing tooth tended to have more metabolic syndrome risk factors. Data were analyzed by descriptive statistics, chi-square test, correlation analysis, logistic regression using SPSS version 18.0. Results : Dental caries revealed 1.146 times higher than the control group. Risk group showed the following results; gingivitis was 1.317 times higher in risk group and 1.612 times in metabolic syndrome. Dental calculus was 1.532 times higher in risk group and 1.557 times in metabolic syndrome. Mssing teeth were 1.976 higher in metabolic syndrome. Conclusions : Metabolic syndrome risk factors had close correlation to poor raol health status. It is necessary to establish the effective plan for the oral health in workers.

Applications of Metabolic Modeling to Drive Bioprocess Development for the Production of Value-added Chemicals

  • Mahadevan, Radhakrishnan;Burgard, Anthony P.;Famili, Iman;Dien, Steve Van;Schilling, Christophe H.
    • Biotechnology and Bioprocess Engineering:BBE
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    • 제10권5호
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    • pp.408-417
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    • 2005
  • Increasing numbers of value added chemicals are being produced using microbial fermentation strategies. Computational modeling and simulation of microbial metabolism is rapidly becoming an enabling technology that is driving a new paradigm to accelerate the bioprocess development cycle. In particular, constraint-based modeling and the development of genome-scale models of industrial microbes are finding increasing utility across many phases of the bioprocess development workflow. Herein, we review and discuss the requirements and trends in the industrial application of this technology as we build toward integrated computational/experimental platforms for bioprocess engineering. Specifically we cover the following topics: (1) genome-scale models as genetically and biochemically consistent representations of metabolic networks; (2) the ability of these models to predict, assess, and interpret metabolic physiology and flux states of metabolism; (3) the model-guided integrative analysis of high throughput 'omics' data; (4) the reconciliation and analysis of on- and off-line fermentation data as well as flux tracing data; (5) model-aided strain design strategies and the integration of calculated biotransformation routes; and (6) control and optimization of the fermentation processes. Collectively, constraint-based modeling strategies are impacting the iterative characterization of metabolic flux states throughout the bioprocess development cycle, while also driving metabolic engineering strategies and fermentation optimization.

Metabolic Flux Distribution in a Metabolically Engineered Escherichia coli Strain Producing Succinic Acid

  • Hong, Soon-Ho;Lee, Sang-Yup
    • Journal of Microbiology and Biotechnology
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    • 제10권4호
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    • pp.496-501
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    • 2000
  • Escherichia cole NZN111, which is known as a pfl ldhA double mutant strin, was metabolically engineered to produce succinic acid by overexpressing malic enzyme into the E. coli controlled by a trc promoter. Fermentation studies were carried out in a LB medium by first growing cells aerobically to an $OD_{600}$ of 5. At this point, 0.01 mM IPTG was added to induce the overexpression of malic enzyme and the agitation speed was gradually lowered. When the culture $OD_{600}$ reached 11, a complete anaerobic condition was achieved by flushing with a $CO_3-H_2$ gas mixture. When NZN111(pTrcML) was cultured at $37^{\circ}C$, the final succinic acid concentration of 2.8 g/l could be obtained after 30 h of anaerobic cultivation. The fermentation results were analyzed by the calculation of metabolic fluxes. Metaolic flux analysis showed that about 85% of phosphoenolpyruvate (PEP) was converted to pyruvate, and further converted to malic acid by malic enzyme.

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구강건강신념과 대사증후군이 지역사회치주치료요구지수에 미치는 영향 (Effect of oral health belief and metabolic syndrome on CPITN)

  • 정명희;김창숙;이경수
    • 한국치위생학회지
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    • 제16권6호
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    • pp.979-991
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    • 2016
  • Objectives: The objective of the study is to investigate the effect of oral health belief and metabolic syndrome on CPITN. Methods: A self-reported questionnaire was completed by 847 adult visitors over 20 years old in Busan from August 1 to December 31, 2011. Except incomplete answers, 776 data were analyzed by t-test, one way ANOVA, and logistic regression analysis using SPSS 20.0 program. The questionnaire comprised general characteristics of the subjects, oral health behavior, and metabolic syndrome related risk factors. Results: There was statistically significant difference of an oral health belief rating between the groups receiving oral health education. Community periodontal index had statistically significant difference depending on smoking and waist circumference. To analyze related factors of community periodontal index, multiple logistic regression analysis was performed. As the final outcome, gender, age, benefit, salience, scaling experience, experience of oral health education, and related factors and metabolic syndrome were shown to impact on significant influence (p<0.05). Conclusions: Thus, To manage the effective oral health, it is necessary to control the factors of metabolic syndrome and oral health.

The Role of High-throughput Transcriptome Analysis in Metabolic Engineering

  • Jewett, Michael C.;Oliveira, Ana Paula;Patil, Kiran Raosaheb;Nielsen, Jens
    • Biotechnology and Bioprocess Engineering:BBE
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    • 제10권5호
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    • pp.385-399
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    • 2005
  • The phenotypic response of a cell results from a well orchestrated web of complex interactions which propagate from the genetic architecture through the metabolic flux network. To rationally design cell factories which carry out specific functional objectives by controlling this hierarchical system is a challenge. Transcriptome analysis, the most mature high-throughput measurement technology, has been readily applied In strain improvement programs in an attempt to Identify genes involved in expressing a given phenotype. Unfortunately, while differentially expressed genes may provide targets for metabolic engineering, phenotypic responses are often not directly linked to transcriptional patterns, This limits the application of genome-wide transcriptional analysis for the design of cell factories. However, improved tools for integrating transcriptional data with other high-throughput measurements and known biological interactions are emerging. These tools hold significant promise for providing the framework to comprehensively dissect the regulatory mechanisms that identify the cellular control mechanisms and lead to more effective strategies to rewire the cellular control elements for metabolic engineering.

Mapping of Carbon Flow Distribution in the Central Metabolic Pathways of Clostridium cellulolyticum: Direct Comparison of Bacterial Metabolism with a Soluble versus an Insoluble Carbon Source

  • DESVAUX, MICKAEL,
    • Journal of Microbiology and Biotechnology
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    • 제14권6호
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    • pp.1200-1210
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    • 2004
  • Metabolic flux analysis was established by adapting previous stoichiometric model developed during growth with cellulose to cell grown with cellobiose for further direct comparison of the bacterial metabolism. In carbon limitation with cellobiose, a shift from acetate-ethanol fermentation to ethanol-lactate fermentation is observed and the pyruvate overflow is much higher than with cellulose. In nitrogen limitation with cellobiose, the cellodextrin and exopolysaccharide overflows are much higher than on cellulose. In carbon and nitrogen saturation with cellobiose, the cellodextrin, exopolysaccharide, and free amino acids overflows reach the highest levels observed but all remain limited on cellulose. By completely shunting the cellulosome, the use of cellobiose allows to reach much higher carbon consumption rates which, in return, highlights the metabolic limitation of C. cellulolyticum. Therefore, the physical nature of the carbon source has a profound impact on the metabolism of C. cellulolyticum and most probably of other cellulolytic bacteria. For cellulolytic bacteria, the use of soluble carbon substrate must carefully be taken into consideration for the interpretation of results. Direct comparison of metabolic flux analysis from cellobiose and cellulose revealed the importance of cellulosome, phosphoglucomutase and pyruvate-ferredoxin oxidoreductase in the distribution of carbon flow in the central metabolism. In the light of these findings, future directions for improvement of cellulose catabolism by this bacterium are discussed.

커피와 대사증후군 : 체계적 문헌고찰 및 메타분석 (Coffee and metabolic syndrome: A systematic review and meta-analysis)

  • 이유진;손자경;장지영;박경
    • Journal of Nutrition and Health
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    • 제49권4호
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    • pp.213-222
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    • 2016
  • Purpose: Coffee is the most frequently consumed food item in South Korea after rice and cabbage. Coffee contains various substances, including caffeine, cafestol, kahweol, chlorogenic acid, and many other known and unknown ingredients with some health benefits. Especially, cumulative evidence has shown that regular coffee use is associated with lower risk of type 2 diabetes, although limited and inconsistent data are available regarding metabolic syndrome. Methods: This study reviewed all available scientific and epidemiologic evidence on coffee consumption, metabolic syndrome, and the association between them. Most epidemiologic research regarding this association was of a cross-sectional design, and a few case-control and cohort studies were available. We conducted meta-analysis with 11 observational studies investigated in Europe, America, and Asia. Summary odds ratios (OR) were calculated using a random-effects model. Results: The overall OR of metabolic syndrome was 0.90 (95% Confidence Interval (CI), 0.81-0.99) for the highest category of coffee intake compared with the lowest intake category. These associations were stronger in populations of US and Europe (OR 0.84, 95% CI 0.76-0.94), whereas no association was observed in the Asian population (OR 1.00, 95% CI 0.81-1.23). Conclusion: The review results indicate that frequent coffee consumption may be beneficial to metabolic syndrome, but the association between coffee consumption and metabolic syndrome may differ by nations or continents.

Development of a food-based index of dietary inflammatory potential for Koreans and its relationship with metabolic syndrome

  • Na, Woori;Yu, Tae Yang;Sohn, Cheongmin
    • Nutrition Research and Practice
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    • 제13권2호
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    • pp.150-158
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    • 2019
  • BACKGROUND/OBJECTIVES: Inflammation is known to be a risk factor for metabolic diseases. This study aimed to develop a Food-based Index of Dietary Inflammatory Potential (FBDI) and examine its association with metabolic biomarkers. SUBJECTS/METHODS: This study analyzed the raw data from the 2012-2014 Korean Genome and Epidemiology Study data of 17,771 people. To analyze the relationship between foods consumed by Koreans and inflammation, we conducted a correlation analysis between 51 food groups and hs-CRP levels. The FBDI was developed from 17 food groups selected by multiple regression method. We examined whether FBDI was associated with metabolic markers (waist circumference, blood pressure, fasting glucose, triglyceride, and HDL-cholesterol) in the 6th Korea National Health and Nutrition Examination Survey (KNHANES). We used binary logistic regression analysis to examine the association. RESULTS: The FBDI model included seven of the anti-inflammatory food groups and three of the pro-inflammatory food groups. The FBDI formula was calculated by multiplying the intake of food group by ${\beta}$-coefficients derived from the multiple regression model based on the correlation analysis. The FBDI was significantly associated with waist circumference (P < 0.001), blood pressure (P < 0.001), triglyceride level (P < 0.001), and HDL-cholesterol (P < 0.001) level among adults aged 20-64 years in the KNHANES. The prevalence of metabolic syndrome was 2.618 times higher in the group with the highest FBDI than in the group with the lowest one (95% confidence interval: 1.778-3.856, P for trend < 0.001). CONCLUSIONS: This study established an FBDI reflecting food intake patterns of Koreans, which showed a significant relationship with the prevalence of metabolic syndrome.