• Title/Summary/Keyword: protein tissues

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Effects of Warming Acupuncture on Ligament Recovery in Injury-induced Rats

  • Heo, Dong-Seok;Geum, Dong-Ho
    • The Journal of Korean Medicine
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    • v.27 no.4
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    • pp.156-161
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    • 2006
  • Object : Warming acupuncture (WA) has been used in Oriental Medicine for the treatment of physical disabilities caused by ligament damage. Here, the effects of WA on injured ligament tissues were investigated using the rat model. Methods : The rats were induced injury on the right hind ankle, and 4 weeks later, WA was given onto the acupoint GB4O(Quixu) of the injury area on a weekly basis for 6 weeks. Main outcome was measured by levels of Erk1/2. Hoechst nuclear staining and collagen staining in the ligament tissue. Result : Levels of active form of Erk1/2 kinase were increased in the injured ligament with WA compared with the control ligament induced injury only, and this change correlated with cell number increases in the ligament by WA. Type III, but not type I, collagen mRNA and protein levels were elevated in the injured ligament treated with WA. Moreover, histological staining showed increased re-organization of collagen fibers in the ligament by WA. Conclusions : The present data suggest that WA performance to the injured ligament may facilitate the healing process via increasing cellular activity.

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A New Assay Method for Spermidine and Spermine Synthases Using Antibody Against MTA

  • Lee, Sung-Ho;Cho, Young-Dong
    • BMB Reports
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    • v.30 no.6
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    • pp.443-447
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    • 1997
  • We have developed a novel method for assays of spermidine and spermine synthase (aminopropyltransferase) activities using antibody against 5'-deoxy-5'-methylthioadenosine (MTA). A new assay is reported here which is based on the observation that MTA is formed as a stoichiometric by-product of the spermidine and spermine synthases reactions. In order to determine MTA, a radioimmunoassay method with sensitivity and rapidity was used. (Lee and Cho, 1997). In this assay, adenine must be added in the reaction mixture, since it effectively inhibits the action of MTA phosphorylase by which MTA is metabolized. This assay is a improvement in term of sensitivity and time saving, compared to the currently used methods. It has a level of sensitivity (100 fmol) sufficient to monitor aminopropyltransferase activities in incubations containing as little as $10{\mu}g$ protein prepared from rat tissue homogenate. The results obtained showed that this method is particularly useful for cultured cells with low enzyme concentration. Moreover, this assay has the advantage which allows studies using alternative substrates (other amines). Spermidine synthase activity was high in rat liver, but low in rat kidney. The activity of spermine synthase was in most rat tissues very low as compared to that of spermidine synthase, but was high in brain.

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Identification of MFGE8 in mesenchymal stem cell secretome as an anti-fibrotic factor in liver fibrosis

  • Jang, Yu Jin;An, Su Yeon;Kim, Jong-Hoon
    • BMB Reports
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    • v.50 no.2
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    • pp.58-59
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    • 2017
  • The beneficial paracrine roles of mesenchymal stem cells (MSCs) in tissue repair have potential in therapeutic strategies against various diseases. However, the key therapeutic factors secreted from MSCs and their exact molecular mechanisms of action remain unclear. In this study, the cell-free secretome of umbilical cord-derived MSCs showed significant anti-fibrotic activity in the mouse models of liver fibrosis. The involved action mechanism was the regulation of hepatic stellate cell activation by direct inhibition of the $TGF{\beta}$/Smad-signaling. Antagonizing the milk fat globule-EGF factor 8 (MFGE8) activity blocked the anti-fibrotic effects of the MSC secretome in vitro and in vivo. Moreover, MFGE8 was secreted by MSCs from the umbilical cord as well as other tissues, including teeth and bone marrow. Administration of recombinant MFGE8 protein alone had a significant anti-fibrotic effect in two different models of liver fibrosis. Additionally, MFGE8 downregulated $TGF{\beta}$ type I receptor expression by binding to ${\alpha}v{\beta}3$ integrin on HSCs. These findings revealed the potential role of MFGE8 in modulating $TGF{\beta}$-signaling. Thus, MFGE8 could serve as a novel therapeutic agent for liver fibrosis.

Melatonin-Induced PGC-1α Improves Angiogenic Potential of Mesenchymal Stem Cells in Hindlimb Ischemia

  • Lee, Jun Hee;Han, Yong-Seok;Lee, Sang Hun
    • Biomolecules & Therapeutics
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    • v.28 no.3
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    • pp.240-249
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    • 2020
  • Despite the therapeutic effect of mesenchymal stem cells (MSCs) in ischemic diseases, pathophysiological conditions, including hypoxia, limited nutrient availability, and oxidative stress restrict their potential. To address this issue, we investigated the effect of melatonin on the bioactivities of MSCs. Treatment of MSCs with melatonin increased the expression of peroxisome proliferator-activated receptor gamma coactivator-1 alpha (PGC-1α). Melatonin treatment enhanced mitochondrial oxidative phosphorylation in MSCs in a PGC-1α-dependent manner. Melatonin-mediated PGC-1α expression enhanced the proliferative potential of MSCs through regulation of cell cycle-associated protein activity. In addition, melatonin promoted the angiogenic ability of MSCs, including migration and invasion abilities and secretion of angiogenic cytokines by increasing PGC-1α expression. In a murine hindlimb ischemia model, the survival of transplanted melatonin-treated MSCs was significantly increased in the ischemic tissues, resulting in improvement of functional recovery, such as blood perfusion, limb salvage, neovascularization, and protection against necrosis and fibrosis. These findings indicate that the therapeutic effect of melatonin-treated MSCs in ischemic diseases is mediated via regulation of PGC-1α level. This study suggests that melatonin-induced PGC-1α might serve as a novel target for MSC-based therapy of ischemic diseases, and melatonin-treated MSCs could be used as an effective cell-based therapeutic option for patients with ischemic diseases.

A Safety Evaluation of Genetically Modified Feedstuffs for Livestock Production; the Fate of Transgenic DNA and Proteins

  • Beever, D.E.;Glenn, K.;Phipps, R.H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.16 no.5
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    • pp.764-772
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    • 2003
  • Two genetic constructs used to confer improved agronomic characteristics, namely herbicide tolerance (HT) in maize and soyabean and insect resistance (Bt) in maize, are considered in respect of feeding to farm livestock, animal performance and the nutritional value and safety of animal products. A review of nucleic acid (DNA) and protein digestion in farm livestock concludes that the frequency of intact transgenic DNA and proteins of GM and non-GM crops being absorbed is minimal/non existent, although there is some evidence of the presence of short fragments of rubisco DNA of non-GM soya in animal tissues. It has been established that feed processing (especially heat) prior to feeding causes significant disruption of plant DNA. Studies with ruminant and non-ruminant farm livestock offered GM feeds demonstrated that animal performance and product composition are unaffected and that there is no evidence of transgenic DNA or proteins of current GM in the products of animals consuming such feeds. On this evidence, current HT and Bt constructs represent no threat to the health of animals, or humans consuming the products of such animals. However as new GM constructs become available it will be necessary to subject these to rigorous evaluation.

Isolation and Characterization of Mucinase Complex Secreted from Vibrio parahaemolyticus

  • Jun, In-Joon;Kim, Yoon-Hee;Kim, Min-Jeong;Hwang, Ho-Soon;Lee, Tae-Ho;Cha, Jae-Ho
    • Journal of Microbiology and Biotechnology
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    • v.13 no.5
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    • pp.731-737
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    • 2003
  • Mucinase complex from V. parahaemolyticus ATCC 17802 was purified 6-fold with 0.4% yield by two sequential steps of Q-Sepharose and Superdex 200HR column chromatographies. Partially purified mucinase complex showed at least 8 times higher mucin-degrading activity than the culture filtrates. The mucinase complex also showed gelatin-and-casein-hydrolyzing activities, which demonstrates that the protein is a complex compound containing several proteases. The optimum pH and temperature of partially purified mucinase complex for mucin degradation was 8.0 and $35^{\circ}C$, respectively. The partially purified mucinase complex showed high cytotoxic activity on vero cells when examined by MTT assay and microscopic observations. Cytotoxicity was significantly increased in proportion to the concentration of the mucinase complex. Mouse experiments revealed that the jejunum, ileum. and large intestinal tissues were damaged by the injection of the mucinase complex. In particular, the reduction of the goblet cells in the large intestine was remarkable. Collectively, these data suggest that the mucinase complex partially purified from V. parahaemolyticus ATCC 17802 contributes to the adhesion and invasion of V. parahaemolyticus into the host intestinal tract.

Antimicrobial Peptides (AMPs): Peptide Structure and Mode of Action

  • Park, Yoon-Kyung;Hahm, Kyung-Soo
    • BMB Reports
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    • v.38 no.5
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    • pp.507-516
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    • 2005
  • Antimicrobial peptides (AMPs) have been isolated and characterized from tissues and organisms representing virtually every kingdom and phylum. Their amino acid composition, amphipathicity, cationic charge, and size allow them to attach to and insert into membrane bilayers to form pores by 'barrel-stave', 'carpet' or 'toroidal-pore' mechanisms. Although these models are helpful for defining mechanisms of AMP activity, their relevance to resolving how peptides damage and kill microorganisms still needs to be clarified. Moreover, many AMPs employ sophisticated and dynamic mechanisms of action to carry out their likely roles in antimicrobial host defense. Recently, it has been speculated that transmembrane pore formation is not the only mechanism of microbial killing by AMPs. In fact, several observations suggest that translocated AMPs can alter cytoplasmic membrane septum formation, reduce cell-wall, nucleic acid, and protein synthesis, and inhibit enzymatic activity. In this review, we present the structures of several AMPs as well as models of how AMPs induce pore formation. AMPs have received special attention as a possible alternative way to combat antibiotic-resistant bacterial strains. It may be possible to design synthetic AMPs with enhanced activity for microbial cells, especially those with antibiotic resistance, as well as synergistic effects with conventional antibiotic agents that lack cytotoxic or hemolytic activity.

Gene repressive mechanisms in the mouse brain involved in memory formation

  • Yu, Nam-Kyung;Kaang, Bong-Kiun
    • BMB Reports
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    • v.49 no.4
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    • pp.199-200
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    • 2016
  • Gene regulation in the brain is essential for long-term plasticity and memory formation. Despite this established notion, the quantitative translational map in the brain during memory formation has not been reported. To systematically probe the changes in protein synthesis during memory formation, our recent study exploited ribosome profiling using the mouse hippocampal tissues at multiple time points after a learning event. Analysis of the resulting database revealed novel types of gene regulation after learning. First, the translation of a group of genes was rapidly suppressed without change in mRNA levels. At later time points, the expression of another group of genes was downregulated through reduction in mRNA levels. This reduction was predicted to be downstream of inhibition of ESR1 (Estrogen Receptor 1) signaling. Overexpressing Nrsn1, one of the genes whose translation was suppressed, or activating ESR1 by injecting an agonist interfered with memory formation, suggesting the functional importance of these findings. Moreover, the translation of genes encoding the translational machineries was found to be suppressed, among other genes in the mouse hippocampus. Together, this unbiased approach has revealed previously unidentified characteristics of gene regulation in the brain and highlighted the importance of repressive controls.

Insulin as a Potent Stimulator of Akt, ERK and Inhibin-βE Signaling in Osteoblast-Like UMR-106 Cells

  • Ramalingam, Mahesh;Kwon, Yong-Dae;Kim, Sung-Jin
    • Biomolecules & Therapeutics
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    • v.24 no.6
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    • pp.589-594
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    • 2016
  • Insulin is a peptide hormone of the endocrine pancreas and exerts a wide variety of physiological actions in insulin sensitive tissues, such as regulation of glucose homeostasis, cell growth, differentiation, learning and memory. However, the role of insulin in osteoblast cells remains to be fully characterized. In this study, we demonstrated that the insulin (100 nM) has the ability to stimulate the phosphorylation of protein kinase B (Akt/PKB) and extracellular signal-regulated kinase (ERK) and the levels of inhibin-${\beta}E$ in the osteoblast-like UMR-106 cells. This insulin-stimulated activities were abolished by the PI3K and MEK1 inhibitors LY294002 and PD98059, respectively. This is the first report proving that insulin is a potential candidate that enables the actions of inhibin-${\beta}E$ subunit of the TGF-${\beta}$ family. The current investigation provides a foundation for the realization of insulin as a potential stimulator in survival signaling pathways in osteoblast-like UMR-106 cells.

New Antisense RNA Systems Targeted Against Plant Pathogens

  • Matousek, J.;Vrba, L.;Kuchar, M.;Pavingerova, D.;Orctova, L.;Ptacek, J.;Schubert, J.;Steger, G.;Beier, H.;Riesner, D.
    • Korean Journal of Plant Tissue Culture
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    • v.27 no.5
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    • pp.379-385
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    • 2000
  • tRNA and 7SL RNA based antisense vehicles were prepared by inserting conserved anti-viral and anti-viroid domains. Anti-PVS coat protein leader sequence (ACPL) and antistructural antihairpin domain of PSTVd (AHII) were inserted in tRNA cassette; anti- zing finger domain of PVS, AHII and anti hop latent viroid ribozyme were inserted in 7SL RNA gene isolated from A. thaliana. These constructs were shown to be transcribed both, in in vitro and in in vivo conditions. However, it followed from our work that closely linked position of PoIII reference genes and PoIIII antisense genes within T-DNA lead to the impairment of RNA expression in transgenic plants. To assay in vivo transcription of antisense genes, hairy root potato cultures were established using h. tumefaciens A4-24 bearing both, Ri plasmid and PoIII-promoterless plant expression vectors with antisense RNA genes. Expression of antisense RNA in transgenic potato tissues was proven by specific RT-PCR reactions.

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