• Title/Summary/Keyword: protein tissues

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Hemocyte-specific Promoter for the Development of Transgenic Silkworm, Bombyx mori

  • Park, Seung-Won;Goo, Tae-Won;Kim, Seong-Ryul;Choi, Gwang-Ho
    • International Journal of Industrial Entomology and Biomaterials
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    • v.25 no.1
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    • pp.111-114
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    • 2012
  • In previous studies we have shown that a sw17255 gene was expressed in hemocyte-specific tissues of the silkworm, Bombyx mori L. (Lepidoptera: Bombycidae). It was verified that the sw17255 core promoter region contains elements that regulate the expression of this gene in hemocyte tissue; the selected promoter region spans nucleotides -1 to -2,112 upstream of the start codon. Each of the luciferase reporter gene expression vectors under the control of 4 different kinds of promoter candidates, (-2,112/-1), (-1,640/-1), (-1,169/-1) and (-579/-1), and the control reporter plasmid DNA, were introduced into B. mori larval coelom by direct injection using a syringe. The promoter candidate [E] (-579/-1) showed more than 1.67 fold transcriptional activity compared to control promoter activity. Higher productivity of an expressed gene in the transgenic silkworm by this promoter combination could be achieved in the near future. The foreign recombinant protein could be easily harvested from the blood of the transgenic silkworm.

Protective Effects of Thiazolo[3,2-b]-1,2,4-Triazoles on Ethanol­Induced Oxidative Stress in Mouse Brain and Liver

  • Aktay Goknur;Tozkoparan Birsen;Ertan Mevlut
    • Archives of Pharmacal Research
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    • v.28 no.4
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    • pp.438-442
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    • 2005
  • A series of 3-[1-(4-(2-methylpropyl) phenyl) ethyl]-1,2,4-triazole-5-thione (I) and its bicyclic condensed derivatives 6-benzylidenethiazolo[3,2-b]-1, 2,4-triazole-5(6H)-ones (IIa-IIf) were investigated for the prevention of ethanol-induced oxidative stress in liver and brain of mice. Administration of ethanol (0.1 mL/mice, p.o.) resulted in a drop of total thiol groups (T-SH) and non-protein thiol groups (NP-SH), and an increase in thiobarbituric acid reactive substances (TBARS) in both liver and brain tissue of mice (p<0.001). Among the compounds investigated (at a dose of 200 mg/kg, p.o.), I and IId ameliorated the peroxidative injury in these tissues effectively. Compounds IIa, IIc and IIe improved the peroxidative tissue injury only in brain. These findings suggest that certain condensed thiazolo-triazole compounds may contribute to the control of ethanol-induced oxidative stress in an organ selective manner.

Cloning and Nucleotide Sequence of a cDNA Encoding the Rat Triosephosphate Isomerase

  • Lee, Kyunglim;Ryu, Jiwon
    • Archives of Pharmacal Research
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    • v.19 no.6
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    • pp.497-501
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    • 1996
  • A gene coding for triosephosphate isomerase (TPI) from a rat skeletal muscle cDNA library was cloned and its nucleotide sequence was determined. The 1, 348-bp cDNA clone contains 24 bp $5^I$ noncoding region, the entire 750 bp coding region corresponding to a protein of 249 amino acids, $547bp 3^I$ noncoding region and part of a poly(A) tail. It also contains a polyadenylation signal, AATAAA, starting from 17 bp upstream of the poly(A) tail. The calculated molecular weight of rat TPI is 27.8 kDa and the net charge is +4. The deduced amino acid sequence from rat TPI CDNA sequence has 93% and 94% homology with that of mouse and human clones, respectively. The amino acids at the residue of Asn12, Lys14, His96, Glu 166, His96, His101, Ala177, Tyr165, Glu13O, Tyr2O9, and Ser212 in catalytic site are completely identical, confirming that the functional residues in TPI proteins are highly conserved throughout evolution. The most profound characteristic of rat TPI enzyme, compared with other TPIs, is that there are five cysteine substitutions at the residue of 21, 27, 159, 195 and 204. A Glu123 instead of Gly was found in rabbit, rhesus, mouse and human sequences. Through the method of RT-PCR, the mRNA transcription level of TPI gene was found to be different among various tissues and was highest in muscle.

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Enhanced Expression of Plasma Glutathione Peroxidase in the Thymus of Mice Treated with TCDD and Its Implication for TCDD-induced Thymic Atrophy

  • Cho, Hyun-Jin;Hahn, Eun-Jin;Hwang, Ju-Ae;Hong, Min-Sun;Kim, Sook-Kyung;Pak, Hye-Ryun;Park, Joo-Hung
    • Molecules and Cells
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    • v.21 no.2
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    • pp.276-283
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    • 2006
  • The potent environmental contaminant, 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), induces thymus atrophy in experimental animals. However, its mechanism of action is not fully understood. To gain insight into its immunosuppressive effect, Balb/c mice were intraperitoneally injected with TCDD ($30{\mu}g/kg$ body weight) and genes regulated by TCDD were identified using cDNA arrays [Park and Lee (2002)]. One of the regulated genes was that for plasma glutathione peroxidase (pGPx). Upon TCDD injection, pGPx mRNA levels in the thymus increased, in parallel with increases in GPx activity and the frequency of anti-human pGPx antibody-reactive cells. pGPX mRNA levels were also moderately up-regulated in the testis and spleen. This is the first report that a particular isotype of the glutathione peroxidase family is regulated by TCDD at both mRNA and protein levels. pGPx is expressed in various tissues in contact with body fluids, and detoxifies hydrogen peroxides and lipid hydroperoxides. It will be of interest to assess the role of pGPx in TCDD-induced thymic atrophy.

TRAF6 Distinctly Regulates Hematopoietic Stem and Progenitors at Different Periods of Development in Mice

  • Kim, Hyekang;Lee, Seungwon;Lee, Seung-Woo
    • Molecules and Cells
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    • v.41 no.8
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    • pp.753-761
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    • 2018
  • Tumor necrosis factor receptor-associated factor 6 (TRAF6) is identified as a signaling adaptor protein that regulates bone metabolism, immunity, and the development of several tissues. Therefore, its functions are closely associated with multiple diseases. TRAF6 is also involved in the regulation of hematopoiesis under steady-state conditions, but the role of TRAF6 in modulating hematopoietic stem and progenitor cells (HSPCs) during the developmental stages remains unknown. Here, we report that the deletion of TRAF6 in hematopoietic lineage cells resulted in the upregulation of HSPCs in the fetal liver at the prenatal period. However, in the early postnatal period, deletion of TRAF6 drastically diminished HSPCs in the bone marrow (BM), with severe defects in BM development and extramedullary hematopoiesis in the spleen being identified. In the analysis of adult HSPCs in a BM reconstitution setting, TRAF6 played no significant role in HSPC homeostasis, albeit it affected the development of T cells. Taken together, our results suggest that the role of TRAF6 in regulating HSPCs is altered in a spatial and temporal manner during the developmental course of mice.

Pulse Exposure to Ethanol Augments Vascular Contractility Through Stress Response

  • Yang, Eun-Kyoung;Kim, In-Kyeom
    • The Korean Journal of Physiology and Pharmacology
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    • v.5 no.1
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    • pp.47-53
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    • 2001
  • Drinking excessive alcohol has been recognized as a risk factor for hypertension. However, the mechanism by which alcohol intake causes hypertension still remains elusive. We tested the hypothesis that ethanol itself acts as a stress factor on vasculature and indirectly modulates vascular contractility. After end of exposure to 1, 2.5 and 5% ethanol for 45 min, rat aortic strips were subjected to contractile responses, immunoblot for Hsp70 and the measurement of levels of myosin light chain phosphorylation. Exposure to 5% ethanol not only augmented contractions to KCl or phenylephrine, but also increased expression of Hsp70 and the levels of myosin light chain phosphorylation. There were no significant differences in contractions produced by $1\;{\mu}mol/L$ phorbol 12,13-dibutyrate, a protein kinase C activator, whether the tissues were exposed to 5% ethanol or not. This is the first report to show that even short exposure to ethanol has a delayed effect to increase vascular smooth muscle contractility through a modulation of thick filament regulation. It may be a mechanism by which ingestion of alcohol induces hypertension.

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Studies on the Experimental Heart and Heart-Lung Transplantation in the Mongrel Dogs for the Purpose of Clinical Application (임상적용을 위한 한국산 잡견에서의 실험적 심장및 심폐 이식술)

  • 이정렬
    • Journal of Chest Surgery
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    • v.25 no.5
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    • pp.458-468
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    • 1992
  • With the aid of extracorporeal circulation, nine dogs underwent orthotopic cardiopulmonary transplantation after preservation of the donor heart in a hypothermic amino acid[glutamate, aspartate] enriched high potassium extracellular solution, and preservation of the donor lung with hypothermic low potassium dextran solution from June 1990 to May 1991. The mean body weights of dogs were 20kg and the recipients` preoperative hematologic and hemodynamic pictures were within normal range except slightly decreased level of albumin and total protein, which was supposed to be due to malnutrition. The following modifications of the original Stanford technique were emphasized: [1] the posterior mediastinum is dissected as little as possible with meticulous hemostasis; [2] the surgical procedure is kept away from the phrenic and vagus nerves; [3] the tracheal anastomosis may be wrapped with recipient`s pulmonary artery flap or surrouding soft tissues. A combination of Cyclosporine, Azathioprine, corticosteroid was used as perioperative immunosuppressive therapy. Postoperatively all recipients could be weaned from extracorporeal circulation, showing favorable vital signs, but within 24 hours, irreversible congetive heart failure, ascites, arrhythmias developed with a mean survival time 13.6$\pm$6.6[n=9, range=6~26] hours. Hemoglobin and platelet counts were significantly[p<0.05] decreased postoperatively, which is thought to be attributed to blood damage by cardiopulmonary bypass and hemodilution. Postmortem finding included multiple subendocardial patch hemorrhage in both atrial and ventricular cavities, pulmonary and liver congestion, and all tracheal anastomoses were intact. Further consideration about quality control of the animal, infection, rejection, the effect of cardiopulmonary bypass on the experimental animal is required to improve the results.

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Calcium Homeostasis and Regulation of Calbindin-D9k by Glucocorticoids and Vitamin D as Bioactive Molecules

  • Choi, Kyung-Chul;Jeung, Eui-Bae
    • Biomolecules & Therapeutics
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    • v.17 no.2
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    • pp.125-132
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    • 2009
  • Calbindin-$D_{9k}$ (CaBP-9k), a cytosolic calcium-binding protein, is expressed in a variety of tissues, i.e., the duodenum, uterus, placenta, kidney and pituitary gland. Duodenal CaBP-9k is involved in intestinal calcium absorption, and is regulated at transcriptional and post-transcriptional levels by 1,25-dihydroxyvitamin D3, the hormonal form of vitamin D, and glucocorticoids (GCs). Uterine CaBP-9k has been implicated in the regulation of myometrial action(s) through modulation of intracellular calcium, and steroid hormones appear to be the main regulators in its uterine and placental regulation. Because phenotypes of CaBP-9k-null mice appear to be normal, other calcium-transporter genes may compensate for its gene deletion and physiological function in knockout mice. Previous studies indicate that CaBP-9k may be controlled in a tissue-specific fashion. In this review, we summarize the current information on calcium homeostasis related to CaBP-9k gene regulation by GCs, vitamin D and its receptors, and its molecular regulatory mechanism. In addition, we present related data from our current research.

Functional Characterization of NtCDPK1 in Tobacco

  • Lee, Sang Sook;Yoon, Gyeong Mee;Rho, Eun Jung;Moon, Eunpyo;Pai, Hyun-Sook
    • Molecules and Cells
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    • v.21 no.1
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    • pp.141-146
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    • 2006
  • We previously showed that NtCDPK1, a tobacco calcium-dependent protein kinase, interacts with and phosphorylates the Rpn3 regulatory subunit of the 26S proteasome, and that both NtCDPK1 and Rpn3 are mainly expressed in rapidly proliferating tissues, including shoot and root meristem. In this study, we examined NtCDPK1 expression in roots using GUS expression in transgenic Arabidopsis plants, and investigated its function in root development by generating transgenic tobacco plants carrying a sense NtCDPK1 transgene. GUS activity was first detected in roots two days after sowing. In later stages, strong GUS expression was detected in the root meristem and elongation zone, as well as the initiation sites and branch points of lateral roots. Transgenic tobacco plants in which NtCDPK1 expression was suppressed were smaller, and their root development was abnormal, with reduced lateral root formation and less elongation. These results suggest that NtCDPK1 plays a role in a signaling pathway regulating root development in tobacco.

Preparation of Collagen/Poly(L-lactic acid) Composite Material for Wound Dressing

  • Lee, Jung-Soo;Kim, Jae-Kyung;Park, So-Ra;Chang, Yoon-Ho
    • Macromolecular Research
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    • v.15 no.3
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    • pp.205-210
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    • 2007
  • Collagen is the major structural protein of connective tissues. It can be used as a prosthetic biomaterial applicable to artificial skin, tendon, ligaments, and collagen implants. The objective of this study is to investigate the possibility of realizing wound dressing medical products by the synthesis of composite materials with collagen and a biodegradable polymer, PLLA, via a surface modification process. Type I collagen was obtained from pig skin by a separation process. The structural characteristics of the extracted collagen were confirmed by SDS-polyacrylamide (PAcr) gel electrophoresis (PAGE) and FTIR. Also, PLLA-g-PAcr was synthesized by the radical polymerization of acrylamide initiated by AIBN in the presence of PLLA. The surface of PLLA was modified by the presence of the acrylamide residues. The structural characteristics of the copolymer were analyzed by FTIR, $^1H-NMR$ and contact angle measurements. The water uptake and WVTR of the collagen/PLLA-g-PAcr composite tended to increase with increasing collagen concentration and with decreasing EDC concentration.