• Title/Summary/Keyword: Functional Model

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Integrated Quantitative Phosphoproteomics and Cell-Based Functional Screening Reveals Specific Pathological Cardiac Hypertrophy-Related Phosphorylation Sites

  • Kwon, Hye Kyeong;Choi, Hyunwoo;Park, Sung-Gyoo;Park, Woo Jin;Kim, Do Han;Park, Zee-Yong
    • Molecules and Cells
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    • v.44 no.7
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    • pp.500-516
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    • 2021
  • Cardiac hypertrophic signaling cascades resulting in heart failure diseases are mediated by protein phosphorylation. Recent developments in mass spectrometry-based phosphoproteomics have led to the identification of thousands of differentially phosphorylated proteins and their phosphorylation sites. However, functional studies of these differentially phosphorylated proteins have not been conducted in a large-scale or high-throughput manner due to a lack of methods capable of revealing the functional relevance of each phosphorylation site. In this study, an integrated approach combining quantitative phosphoproteomics and cell-based functional screening using phosphorylation competition peptides was developed. A pathological cardiac hypertrophy model, junctate-1 transgenic mice and control mice, were analyzed using label-free quantitative phosphoproteomics to identify differentially phosphorylated proteins and sites. A cell-based functional assay system measuring hypertrophic cell growth of neonatal rat ventricle cardiomyocytes (NRVMs) following phenylephrine treatment was applied, and changes in phosphorylation of individual differentially phosphorylated sites were induced by incorporation of phosphorylation competition peptides conjugated with cell-penetrating peptides. Cell-based functional screening against 18 selected phosphorylation sites identified three phosphorylation sites (Ser-98, Ser-179 of Ldb3, and Ser-1146 of palladin) displaying near-complete inhibition of cardiac hypertrophic growth of NRVMs. Changes in phosphorylation levels of Ser-98 and Ser-179 in Ldb3 were further confirmed in NRVMs and other pathological/physiological hypertrophy models, including transverse aortic constriction and swimming models, using site-specific phospho-antibodies. Our integrated approach can be used to identify functionally important phosphorylation sites among differentially phosphorylated sites, and unlike conventional approaches, it is easily applicable for large-scale and/or high-throughput analyses.

A Comparison of the Density Functional Theory Based Methodologies for the Triplet Excited State of 𝛑-Conjugated Molecules: Time-Dependent DFT (TD-DFT), TD-DFT within Tamm-Dancoff Approximation (TDA-DFT), and Spin-Unrestricted DFT (UDFT) (𝛑-공액계 분자의 삼중항 여기 상태에 대한 Density Functional Theory (DFT)에 기반한 계산 방법들의 비교: 시간-의존 DFT (TD-DFT), Tamm-Dancoff 근사법을 적용한 DFT (TDA-DFT), 스핀-비제한 DFT (UDFT))

  • Ahn, Chang Hwan;Kim, Dongwook
    • Journal of the Korean Chemical Society
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    • v.63 no.2
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    • pp.73-77
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    • 2019
  • We compared methodologies based on the density functional theory (DFT), e.g., time-dependent DFT (TD-DFT), TD-DFT within Tamm-Dancoff approximation (TDA-DFT), and spin-unrestricted DFT (UDFT), that are usually employed to optimize the geometries of ${\pi}$-conjugated molecules in their lowest lying triplet excited ($T_1$) state. As a model system for ${\pi}$-conjugated molecules, we employed 1,2,3,4,5-pentacyano-6-phenyl-benzene. In conjunction with 6-31G(d) basis sets, we made use of gap-tuned range-separated ${\omega}B97X$ functional which is often employed recently in the calculations of molecular excited states. Near the equilibrium geometries, we found that the important difference between the geometries derived at UDFT level and those at TD-DFT or TDA-DFT methods: more stable ground-state energies but higher triplet excitation energies for UDFT derived geometries. In the studies, we discuss such differences in more detail.

Purification of ginseng rare sapogenins 25-OH-PPT and its hypoglycemic, antiinflammatory and lipid-lowering mechanisms

  • Xu, Jing;Liu, Hairong;Su, Guangyue;Ding, Meng;Wang, Wei;Lu, Jincai;Bi, Xiuli;Zhao, Yuqing
    • Journal of Ginseng Research
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    • v.45 no.1
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    • pp.86-97
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    • 2021
  • Background: Panax ginseng Meyer has been used as a nourishing edible herb in East Asia for thousands of years. 25-OH-PPT was first discovered as a natural rare triterpenoid saponin in ginseng stems and leaves by our group. Research found that it showed strong inhibitory effects on α-glucosidase and protein tyrosine phosphatase 1B, and protected cardiocytes (H9c2) through PI3K/Akt pathway. Methods: In the research, in order to optimize the 25-OH-PPT enrichment process, optimal macroporous resins and optimal purification conditions were studied. Meanwhile, the hypoglycemic effect and mechanism of 25-OH-PPT were evaluated by using STZ to establish insulin-dependent diabetic mice and the spontaneous type 2 diabetes DB/DB mice. Results and Conclusion: Research found that 25-OH-PPT can reduce blood glucose and enhance glucose tolerance in STZ model mice. It increases insulin sensitivity by upregulating GLUT4 and AMPK in skeletal muscle, and activating insulin signaling pathways. In DB/DB mice, 25-OH-PPT achieves hypoglycemic effects mainly by activating the insulin signaling pathway. Meanwhile, through the influence of liver inflammatory factors and lipids in serum, it can be seen that 25-OH-PPT has obvious anti-inflammatory and lipid-lowering effects. These results provide new insights into the study of ginseng as a functional food.

In silico characterisation, homology modelling and structure-based functional annotation of blunt snout bream (Megalobrama amblycephala) Hsp70 and Hsc70 proteins

  • Tran, Ngoc Tuan;Jakovlic, Ivan;Wang, Wei-Min
    • Journal of Animal Science and Technology
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    • v.57 no.12
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    • pp.44.1-44.9
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    • 2015
  • Background: Heat shock proteins play an important role in protection from stress stimuli and metabolic insults in almost all organisms. Methods: In this study, computational tools were used to deeply analyse the physicochemical characteristics and, using homology modelling, reliably predict the tertiary structure of the blunt snout bream (Ma-) Hsp70 and Hsc70 proteins. Derived three-dimensional models were then used to predict the function of the proteins. Results: Previously published predictions regarding the protein length, molecular weight, theoretical isoelectric point and total number of positive and negative residues were corroborated. Among the new findings are: the extinction coefficient (33725/33350 and 35090/34840 - Ma-Hsp70/ Ma-Hsc70, respectively), instability index (33.68/35.56 - both stable), aliphatic index (83.44/80.23 - both very stable), half-life estimates (both relatively stable), grand average of hydropathicity (-0.431/-0.473 - both hydrophilic) and amino acid composition (alanine-lysine-glycine/glycine-lysine-aspartic acid were the most abundant, no disulphide bonds, the N-terminal of both proteins was methionine). Homology modelling was performed by SWISS-MODEL program and the proposed model was evaluated as highly reliable based on PROCHECK's Ramachandran plot, ERRAT, PROVE, Verify 3D, ProQ and ProSA analyses. Conclusions: The research revealed a high structural similarity to Hsp70 and Hsc70 proteins from several taxonomically distant animal species, corroborating a remarkably high level of evolutionary conservation among the members of this protein family. Functional annotation based on structural similarity provides a reliable additional indirect evidence for a high level of functional conservation of these two genes/proteins in blunt snout bream, but it is not sensitive enough to functionally distinguish the two isoforms.

Functional Genomic Approaches Using the Nematode Caenorhabditis elegans as a Model System

  • Lee, Jun-Ho;Nam, Seung-Hee;Hwang, Soon-Baek;Hong, Min-Gi;Kwon, Jae-Young;Joeng, Kyu-Sang;Im, Seol-Hee;Shim, Ji-Won;Park, Moon-Cheol
    • BMB Reports
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    • v.37 no.1
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    • pp.107-113
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    • 2004
  • Since the completion of the genome project of the nematode C. elegans in 1998, functional genomic approaches have been applied to elucidate the gene and protein networks in this model organism. The recent completion of the whole genome of C. briggsae, a close sister species of C. elegans, now makes it possible to employ the comparative genomic approaches for identifying regulatory mechanisms that are conserved in these species and to make more precise annotation of the predicted genes. RNA interference (RNAi) screenings in C. elegans have been performed to screen the whole genome for the genes whose mutations give rise to specific phenotypes of interest. RNAi screens can also be used to identify genes that act genetically together with a gene of interest. Microarray experiments have been very useful in identifying genes that exhibit co-regulated expression profiles in given genetic or environmental conditions. Proteomic approaches also can be applied to the nematode, just as in other species whose genomes are known. With all these functional genomic tools, genetics will still remain an important tool for gene function studies in the post genome era. New breakthroughs in C. elegans biology, such as establishing a feasible gene knockout method, immortalized cell lines, or identifying viruses that can be used as vectors for introducing exogenous gene constructs into the worms, will augment the usage of this small organism for genome-wide biology.

An 8-wk, randomized, double-blind, placebo-controlled clinical trial for the antidiabetic effects of hydrolyzed ginseng extract

  • Park, Soo-Hyun;Oh, Mi-Ra;Choi, Eun-Kyung;Kim, Min-Gul;Ha, Ki-Chan;Lee, Seung-Kwon;Kim, Young-Gon;Park, Byung-Hyun;Kim, Dal-Sik;Chae, Soo-Wan
    • Journal of Ginseng Research
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    • v.38 no.4
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    • pp.239-243
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    • 2014
  • Background: To investigate the antidiabetic effects of hydrolyzed ginseng extract (HGE) for Korean participants in an 8-wk, randomized, double-blinded, placebo-controlled clinical trial. Methods: Impaired fasting glucose participants [fasting plasma glucose (FPG) ${\geq}5.6mM$ or < 6.9mM who had not been diagnosed with any disease and met the inclusion criteria were recruited for this study. The 23 participants were randomly divided into either the HGE (n = 12, 960 mg/d) or placebo (n = 11) group. Outcomes included measurements of efficacy (FPG, postprandial glucose, fasting plasma insulin, postprandial insulin, homeostatic model assessment-insulin resistance, and homeostatic model assessment-${\beta}$) and safety (adverse events, laboratory tests, electrocardiogram, and vital signs). Results: After 8 wk of HGE supplementation, FPG and postprandial glucose were significantly decreased in the HGE group compared to the placebo group. No clinically significant changes in any safety parameter were observed. Our study revealed that HGE is a potent antidiabetic agent that does not produce noticeable adverse effects. Conclusion: HGE supplementation may be effective for treating impaired fasting glucose individuals.

Effects of Mungbean Flour Level in Combination with Microbial Transglutaminase on Physicochemical and Textural Properties of Low-salt Pork Model Sausages

  • Lee, Hong Chul;Chin, Koo Bok
    • Food Science of Animal Resources
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    • v.33 no.2
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    • pp.221-228
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    • 2013
  • This study was performed to evaluate the effects of various levels of mungbean flour (MF) (0-2.4%) on the quality characteristics of pork model sausages (PMS) in experiment 1 and also select the optimum level of MF to enhance the water retention and gelling properties of low-salt PMS (LSPMS) with or without microbial transglutaminase (MTGase) in experiment 2. In experiment 1, the addition of MF did not affect pH, chemical compositions (fat and moisture contents), color values, and functional properties (expressible moisture, EM (%) and cooking yield, CY (%)) of PMS. However, the addition of MF increased the chewiness of PMS and hardness if the mungbean flour at the level of more than 1.2% was incorporated. Since the interaction between the microbial transglutaminase (MTGase) treatment and MF level was not significant (p>0.05), data were pooled by different factors (MTGase treatment and MF level) in experiment 2. MF improved the water binding ability and textural springiness of LSPMS. On the other hand, MTGase treatment decreased the pH and cooking yield (%) of LSPMS, but increased most textural properties. In conclusion, the addition of MF could enhance the water retention and textural properties of PMS and LSPMS, regardless of MTGase, when it was added to over 1.2%. Based on these results, mungbean protein may interact with MTGase on the low-salt comminuted meat systems. Therefore, further study might be needed to understand the mechanisms of interaction between MTGase and functional components induced from MF.

Multi-dimensional Factors related to the Functional state of Acute Ischemic Stroke Patients - For Patients Visiting a Hospital in Gangwon-do (급성 허혈성 뇌졸중 환자의 퇴원시 기능 상태와 관련된 다차원적 요인 분석 - 강원도 소재 일개 병원의 환자를 대상으로)

  • Song, Hyun-Ju;Park, Ju-Hyun;Dong, Jae-Yong;Lee, Kwang-Soo
    • The Korean Journal of Health Service Management
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    • v.12 no.2
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    • pp.125-134
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    • 2018
  • Objectives : The purpose of this study was to analyze factors related to the functional state of stroke patients after discharge from hospital. Methods : The data was provided from a hospital in Wonju, Gangwon-do. The subjects of the analysis were those who were admitted to the emergency room due to stroke from July to December 2016. The dependent variable was the patient's functional status as measured by the modified Rankin Scale(mRS). Independent variables were demographic factors (age, sex, and marriage status), transportation and distance factors (transportation, travel distance), inpatient factors (lengths of stay, Charlson Comorbidity Index (CCI), Tissue plasminogen activator, National Institute of Health Stroke Scale (NIHSS). Hierarchial regression analysis was applied for the analysis. Results : In the hierarchical regression analysis, Model 3, including socio-demographic factors, transportation, distance factors, and inpatient factors, was the best fitted model. It showed that functional status of stroke patients was positively associated with age, length of stay, CCI, NIHSS, and negatively associated with unmarried status. Conclusions : Results indicated that management of stroke requires care from the pre-disease stage, and a customized education program policy is needed for high-risk stroke patients who are older and have comorbid illness.

Effects of Product Attributes of Franchise Lunch Box on Consumer's Perceived Value and Repurchase Intention (프랜차이즈 도시락 제품속성이 지각된 가치와 재구매 의도에 미치는 영향 )

  • Eui-Yeon, LEE;Kil-Suk, AHN
    • The Korean Journal of Franchise Management
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    • v.14 no.1
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    • pp.39-55
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    • 2023
  • Purpose: This study is designed to grasp how consumers' beliefs about the lunch box attributes such as healthiness, diversity, price, hygiene, and package eco-friendliness affect consumers' perceived value and purchasing behavior. Research design, data, and methodology: The data were collected from 615 respondents composed of men and women aged 20 to 59 who purchased lunch box products and were sampled using the quota sampling method. An online survey was conducted. The data were analyzed using the measurement model and structural equation model assessment with SPSS 24 and SmartPLS 4.0. Results: First, healthiness, diversity, hygiene, and the price had a significant positive effect on the perceived functional value of consumers, but package eco-friendliness did not significantly affect functional value. On the other hand, healthiness, diversity, price, and package eco-friendliness were found to have a positive impact on consumers' perceived emotional value, but hygiene was found to have no significant effect on emotional value. It was found that functional and emotional values significantly positively affected repurchase intention. Conclusions: This study confirmed that consumers' belief in the product attributes of franchise lunch boxes influenced consumers' perceived value, which thus formed the attitude of consumers and showed a relationship that affects repurchase intention. At the end of this paper, the managerial implications for the franchise lunch box brand, the limitations of this study, and future research directions were presented.