• Title/Summary/Keyword: Myosin light chain

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Force-mediated proinvasive matrix remodeling driven by tumor-associated mesenchymal stem-like cells in glioblastoma

  • Lim, Eun-Jung;Suh, Yongjoon;Kim, Seungmo;Kang, Seok-Gu;Lee, Su-Jae
    • BMB Reports
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    • v.51 no.4
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    • pp.182-187
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    • 2018
  • In carcinoma, cancer-associated fibroblasts participate in force-mediated extracellular matrix (ECM) remodeling, consequently leading to invasion of cancer cells. Likewise, the ECM remodeling actively occurs in glioblastoma (GBM) and the consequent microenvironmental stiffness is strongly linked to migration behavior of GBM cells. However, in GBM the stromal cells responsible for force-mediated ECM remodeling remain unidentified. We show that tumor-associated mesenchymal stem-like cells (tMSLCs) provide a proinvasive matrix condition in GBM by force-mediated ECM remodeling. Importantly, CCL2-mediated Janus kinase 1 (JAK1) activation increased phosphorylation of myosin light chain 2 in tMSLCs and led to collagen assembly and actomyosin contractility. Collectively, our findings implicate tMSLCs as stromal cells providing force-mediated proinvasive ECM remodeling in the GBM microenvironment, and reminiscent of fibroblasts in carcinoma.

The Effect of Sagunja Decoction on Functional Dyspepsia - A Systematic Review and Meta-Analysis (기능성 소화불량에 대한 사군자탕의 치료효과 - 체계적 문헌고찰과 메타분석)

  • Kim, Kyong-lim;Je, Yu-ran;Kim, Kyoung-min
    • The Journal of Internal Korean Medicine
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    • v.42 no.3
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    • pp.259-278
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    • 2021
  • Objectives: This study examines the effect of Sagunja-tang on functional dyspepsia (FD) through a systematic review and meta-analysis of a randomized controlled trial (RCT). Methods: A search for RCTs that tested the effect of Sagunja-tang on functional dyspepsia was conducted in Medline, Embase, PubMed, CENTRAL, CiNii, CNKI, NDSL, RISS, OASIS, and KISS databases on November 8, 2020, with no limit on the year of publication. A meta-analysis was performed by synthesizing the findings, including total efficiency, clinical symptom score, myosin light-chain kinase (MLCK) level (pg/mL, and gastric half-emptying time (min). RevMan 5.4.1 software was used for data analysis. The quality of the literature was evaluated using Cochrane's risk of bias (RoB) tool. Results: A total of 14 RCTs met the selection criteria. As a result of the meta-analysis, the treatment group had higher total efficacy and MLCK levels (gastric antrum, jejunum) than the control group, and the clinical symptom score and gastric half-emptying time were lower. However, due to the low quality of the included RCT and the small sample size, the results may be slightly biased.

Pathophysiology and protective approaches of gut injury in critical illness

  • Jung, Chang Yeon;Bae, Jung Min
    • Journal of Yeungnam Medical Science
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    • v.38 no.1
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    • pp.27-33
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    • 2021
  • The gut is a complex organ that has played an important role in digestion, absorption, endocrine functions, and immunity. The gut mucosal barriers consist of the immunologic barrier and nonimmunologic barrier. During critical illnesses, the gut is susceptible to injury due to the induction of intestinal hyperpermeability. Gut hyperpermeability and barrier dysfunction may lead to systemic inflammatory response syndrome. Additionally, gut microbiota are altered during critical illnesses. The etiology of such microbiome alterations in critical illnesses is multifactorial. The interaction or systemic host defense modulation between distant organs and the gut microbiome is increasingly studied in disease research. No treatment modality exists to significantly enhance the gut epithelial integrity, permeability, or mucus layer in critically ill patients. However, multiple helpful approaches including clinical and preclinical strategies exist. Enteral nutrition is associated with an increased mucosal barrier in animal and human studies. The trophic effects of enteral nutrition might help to maintain the intestinal physiology, prevent atrophy of gut villi, reduce intestinal permeability, and protect against ischemia-reperfusion injury. The microbiome approach such as the use of probiotics, fecal microbial transplantation, and selective decontamination of the digestive tract has been suggested. However, its evidence does not have a high quality. To promote rapid hypertrophy of the small bowel, various factors have been reported, including the epidermal growth factor, membrane permeant inhibitor of myosin light chain kinase, mucus surrogate, pharmacologic vagus nerve agonist, immune-enhancing diet, and glucagon-like peptide-2 as preclinical strategies. However, the evidence remains unclear.

Muscle differentiation induced up-regulation of calcium-related gene expression in quail myoblasts

  • Park, Jeong-Woong;Lee, Jeong Hyo;Kim, Seo Woo;Han, Ji Seon;Kang, Kyung Soo;Kim, Sung-Jo;Park, Tae Sub
    • Asian-Australasian Journal of Animal Sciences
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    • v.31 no.9
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    • pp.1507-1515
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    • 2018
  • Objective: In the poultry industry, the most important economic traits are meat quality and carcass yield. Thus, many studies were conducted to investigate the regulatory pathways during muscle differentiation. To gain insight of muscle differentiation mechanism during growth period, we identified and validated calcium-related genes which were highly expressed during muscle differentiation through mRNA sequencing analysis. Methods: We conducted next-generation-sequencing (NGS) analysis of mRNA from undifferentiated QM7 cells and differentiated QM7 cells (day 1 to day 3 of differentiation periods). Subsequently, we obtained calcium related genes related to muscle differentiation process and examined the expression patterns by quantitative reverse-transcription polymerase chain reaction (qRT-PCR). Results: Through RNA sequencing analysis, we found that the transcription levels of six genes (troponin C1, slow skeletal and cardiac type [TNNC1], myosin light chain 1 [MYL1], MYL3, phospholamban [PLN], caveolin 3 [CAV3], and calsequestrin 2 [CASQ2]) particularly related to calcium regulation were gradually increased according to days of myotube differentiation. Subsequently, we validated the expression patterns of calcium-related genes in quail myoblasts. These results indicated that TNNC1, MYL1, MYL3, PLN, CAV3, CASQ2 responded to differentiation and growth performance in quail muscle. Conclusion: These results indicated that calcium regulation might play a critical role in muscle differentiation. Thus, these findings suggest that further studies would be warranted to investigate the role of calcium ion in muscle differentiation and could provide a useful biomarker for muscle differentiation and growth.

Proteome analysis of the m. longissimus dorsi between fattening stages in Hanwoo steer

  • Kim, Nam-Kuk;Lee, Seung-Hwan;Cho, Yong-Min;Son, Eun-Suk;Kim, Kyung-Yun;Lee, Chang-Soo;Yoon, Du-Hak;Im, Seok-Ki;Oh, Sung-Jong;Park, Eung-Woo
    • BMB Reports
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    • v.42 no.7
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    • pp.433-438
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    • 2009
  • The objective of this study was to identify proteins in the m. longissimus dorsi between early (12 months of age) and late (27 months of age) fattening stages of Hanwoo (Korean cattle) steers. Using two-dimensional electrophoresis and mass spectrometry, 8 proteins of 11 differentially expressed spots between the 12 and 27 month age groups were identified in the loin muscle. Among those that were differentially expressed, zinc finger 323 and myosin light chain were highly expressed in late-fattening stage, and two catabolic enzymes, triosephosphate isomerase (TPI) and succinate dehydrogenase (SDH) were expressed more in the early versus the late-fattening stage. In particular, the quantification of TPI and SDH by immunoblotting correlated well with fat content. Our data suggested that TPI and SDH are potential candidates as markers and their identification provides new insight into the molecular mechanisms and pathways associated with intramuscular fat contents of bovine skeletal muscle.

AMPK γ is Required for Maintaining Epithelial Cell Structure and Polarity (AMPK γ 유전자의 표피세포극성 유지기능 규명)

  • Koh, Hyong-Jong
    • Journal of Life Science
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    • v.21 no.5
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    • pp.621-626
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    • 2011
  • AMP-activated protein kinase (AMPK), a heterotrimeric complex comprising a catalytic ${\alpha}$ subunit and regulatory ${\beta}$ and ${\gamma}$ subunits, has been primarily studied as a major metabolic regulator in various organisms, but recent genetic studies discover its novel physiological functions. The first animal model with no functional AMPK ${\gamma}$ subunit gene was generated by using Drosophila genetics. AMPK ${\gamma}$ flies demonstrated lethality with severe defects in cuticle formation. Further histological analysis found that deletion of AMPK ${\gamma}$ causes severe defects in cell polarity in embryo epithelia. The phosphorylation of nonmuscle myosin regulatory light chain (MRLC), a critical regulator of epithelial cell polarity, was also diminished in AMPK ${\gamma}$ embryo epithelia. These defects in AMPK ${\gamma}$ mutant epithelia were successfully restored by over-expression of AMPK ${\gamma}$. Collectively, these results suggested that AMPK ${\gamma}$ is a critical cell polarity regulator in metazoan development.

The Relationship of the L-type $Ca^{2+}$ Channel on the Depolarization-and Depletion of SR $Ca^{2+}$ -induced Smooth Muscle Contraction and Intracellular $Ca^{2+}$ Mobilization (탈분극과 근장그물 내 $Ca^{2+}$ 고갈-유도 평활근의 수축 및 세포 내 $Ca^{2+}$ 변동에 관여하는 L-형 $Ca^{2+}$ 통로의 상관성)

  • Kim, Jung-Hwan
    • The Journal of Korean Physical Therapy
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    • v.19 no.5
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    • pp.65-76
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    • 2007
  • Purpose: It is generally accepted that smooth muscle contraction is triggered by intracellular $Ca^{2+}$ ($[Ca^{2+}]_i$) released from intracellular $Ca^{2+}$ stores such as sarcoplasmic teticulum (SR) and from the extracellular space. The increased $[Ca^{2+}]^i$ can phosphorylate the 20,000 dalton myosin light chain $(MLC_{20})$ by activating MLC kinase (MLCK), and this initiates smooth muscle contraction. In addition to the $[Ca^{2+}]_i$MACK-tension pathway, a number of intracellular signal molecules, including mitogen-activated protein kinase (MAPK), protein kinase C (PKC) and others, play important roles in the regulation of smooth muscle contraction. However, the mechanisms regulating contraction of depletion of SR $Ca^{2+}$ in mouse gastric smooth muscle strips is not still clear. Methods: To investigate the rotes of $Ca^{2+}$ influx and SR $Ca^{2+}$ release channel on gastric motility, isometric contraction and $[Ca^{2+}]_i$ were examined in mouse gastric smooth muscle strips. Results: High KCl, ryanodine, an activator of $Ca^{2+-}$induced $Ca^{2+}$ release channel, and cyclopiazonic acid (CPA), an inhibitor of SR $Ca^{2+-}$ATPase evoked a sustained increase in muscle contraction and $[Ca^{2+}]_i$. These increases induced by high KCl, ryanodine, and CPA were partially blocked by application of verapamil ($10{\mu}M$), a L-type $Ca^{2+}$ channel inhibitor. Additionally, in $Ca^{2+-}$free solution (1 mM EGTA), ryanodine and CPA had no effect contraction and $[Ca^{2+}]_i$ in fundic muscle strips. Conclusion: These results that extracellular $Ca^{2+}$ influx and depletion of SR trigger $Ca^{2+}$ influx through verapamil-sensitive $Ca^{2+}$ channel, and extracellular and SR $Ca^{2+}$ store may functionally involve in the subcellular $Ca^{2+}$ mobilization in mouse gastric muscle.

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Changes in Cytosolic $Ca^{2+}$ but not in cGMP Contents May be more Important to Nitric Oxide-Mediated Relaxation in Depolarized Vascular Smooth Muscle

  • Lee, Hyun-Seok;Chang, Ki-Churl
    • The Korean Journal of Physiology and Pharmacology
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    • v.2 no.1
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    • pp.63-68
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    • 1998
  • Nitric oxide (NO)-mediated relaxation in vascular smooth muscle involves not only activation of guanylate cyclase but also hyperpolarization of the membrane. It has been shown that depolarization decreases the [$Ca^{2+}$] sensitivity of myosin light chain kinase in arterial smooth muscle, and nitric oxide (NO)-mediated relaxation was attenuated in this situation. However, why potassium inhibits or attenuates the action of EDRF/NO is not clear. Therefore, we investigated the magnitude of relaxation and cGMP contents using measures known to release NO, such as photorelaxation, photo activated NO-mediated relaxation, and NO-donor (SNP)-mediated relaxation in porcine coronary arterial rings in which contractile conditions were made by different degree of depolarization, i.e., contraction in response to U46619 or U46619 plus KCl. In all cases, the magnitude of relaxation was significantly greater (P<0.05) in U46619-contracted rings than in U46619+KCl-contracted ones. Although accumulation of cGMP was evident with three measures employed in the present study, no difference was found in cGMP contents between U46619 and U46619+KCl conditions, indicating that the diminished relaxation in KCl containing solution is cGMP-independent mechanism(s). To understand this further, cytosolic $Ca^{2+}$ changes due to NO were compared in rat thoracic aorta by exploiting photoactivated NO using streptozotocin (STZ) that was contracted with either NE or KCl. Fura-3 $[Ca]_{cyt}$ signal caused by NO was small and transient in high $K^+$-, but large and sustained in NE-contracted aorta. The inhibitory potency of STZ expressed in terms of $IC_{50}$ was 5.14 and 3.88 ${\mu}M$ in NE and in high $K^+$, respectively. These results suggest that modification of the cellular mobilization of $Ca^{2+}$ rather than cGMP levels may be an important mechanism for the NO-mediated relaxation when vascular membrane is depolarized, such as atherosclerosis and hypertension.

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Developmental Proteomic Profiling of Porcine Skeletal Muscle during Postnatal Development

  • Kim, Nam-Kuk;Lim, Jong-Hyun;Song, Min-Jin;Kim, Oun-Hyun;Park, Beom-Young;Kim, Myung-Jick;Hwang, In-Ho;Lee, Chang-Soo
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.10
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    • pp.1612-1617
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    • 2007
  • In this study, we have compared the skeletal muscle proteome at various stages of porcine postnatal development. Korean native pigs were divided into five postnatal stages of 30, 70, 130, 170 and 300 d and their loin muscles were analyzed for muscle proteome by using two-dimensional electrophoresis and mass spectrometry. We found 5 proteins showing a consistent pattern during skeletal muscle growth. Four proteins were identified as myosin light chain 1 slow-twitch (MLC1sa) isoform, troponin T, triosephosphate isomerase (TIP) and DJ-1 protein. The remaining protein was not identified. Two muscle fiber proteins of MLC1sa isoform and troponin T showed a high expression level at an early postnatal stage and then their levels were decreased markedly during growth stages. On the other hand, the expression of TIP and DJ-1 protein, which are well known as catalysis enzyme and antioxidant-related protein, respectively, were linearly increased during growth stages. Thus, the stage-related muscle proteins may be useful as parameters for understanding the developmental characteristics of biochemical and physiological properties in Korean native pig skeletal muscle.

Proteomic Comparison between Japanese Black and Holstein Cattle by Two-dimensional Gel Electrophoresis and Identification of Proteins

  • Ohsaki, H.;Okada, M.;Sasazaki, S.;Hinenoya, T.;Sawa, T.;Iwanaga, S.;Tsuruta, H.;Mukai, F.;Mannen, H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.5
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    • pp.638-644
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    • 2007
  • Differences of meat qualities between Japanese Black and Holstein have been known in Japan, however, the causative proteins and/or the genetic background have been unclear. The aim of this study was to identify candidate proteins causing differences of the meat qualities between the two breeds. Using technique of two-dimensional gel electrophoresis, protein profiling was compared from samples of the longissimus dorsi muscle and subcutaneous adipose tissue. Five protein spots were observed with different expression levels between breeds. By using LC-MS/MS analysis and Mascot program, three of them were identified as ankyrin repeat protein 2, phosphoylated myosin light chain 2 and mimecan protein. Subsequently, we compared the DNA coding sequences of three proteins between breeds to find any nucleotide substitution. However, there was no notable mutation which could affect pI or molecular mass of the proteins. The identified proteins may be responsible for different characteristics of the meat qualities between Japanese Black and Holstein cattle.