• Title/Summary/Keyword: UTP

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Studies on Enzymatic Characteristic′s of Adenylate Kinase from Baker′s Yeast (제빵효모 Adenylate Kinase의 효소학적 특성에 관하여)

  • ;Takahisa Ohta;Hiroshi Sakai
    • Microbiology and Biotechnology Letters
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    • v.12 no.4
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    • pp.277-283
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    • 1984
  • In the forward reaction (ADP formation) of the adenylate kinase from baker's yeast, dissociation constants from binary complexes are higher by a factor of about 4 times then those from at ternary complexes. In the reverse reaction, dissociation constants from the binary complexes are 2 times higher then those from the ternary complexes. The enzyme showed activities against various nucleotide triphospate in following orders; ATP 100, UTP 18, ITP 9 and GTP 5, of the necleotide monophosphate. only dAMP showed 33% activity of that AMP as phosphate acceptor. Divalent cations were required in enzyme reaction in following orders; $Mg^{2+}$ 100, Co$^{2+}$ 57, Mn$^{2+}$ 54, $Ca^{2+}$ 51, Ni$^{2+}$ 10 and Sn$^{2+}$ 6. AMP, as a substrate inhibitor, competitively inhibited the adenylate kinase at pH 7.2 or 8.0. Inhibition constants of the enzyme showed greater dependence on the pH of the reaction mixture, which was the lower Ki values under higher pH. Adenosine pentaphospho adenosine was competive inhibitor to the enzyme against all substrate, and it showed the same Ki values, 2.9mM. Further, PEP was competive inhibitor with respect to AMP and non-competive inhibitor with respect to MgATP. Adenylate kinase from bakers yeast was similar to mitochondrial type of animal in the contents of aianine, leucine and asparagine or asparatic acid differing from muscle type enzyme. Based on the results and observation, characteristic of yeast adenylate kinase resembled the adenylate kinase of mitochondrial type from animals. Further, difference of characteristics in adenylate kinasa depending upon the workers might be due to the difference of strain used.

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Effect of treadmill exercise on apoptosis in the retinas of streptozotocin-induced diabetic rats (트레드밀 운동이 streptozotocin에 의해 유발된 당뇨 쥐의 망막 신경세포 사멸에 미치는 영향)

  • Kim, D.Y.;Jung, S.Y.;Kim, T.W.;Sung, Y.H.
    • Exercise Science
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    • v.21 no.3
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    • pp.289-298
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    • 2012
  • In the present study, we investigated the effect of treadmill exercise on apoptotic neuronal cell death in the retinas of streptozotocin-induced diabetic rats. Twenty-eight male Sprague-Dawley rats were used for this study. The animals were divided into four groups(n = 7 in each group):(1) control group, (2) exercise group, (3) diabetes-induced group, (4) diabetes-induced and exercise group. Diabetes mellitus(DM) was induced by intraperitoneal injection of streptozotocin. The rats in the exercise groups were forced to run on the treadmill for 30 minutes once a day, five times per a week, during 12 weeks. In this study, a terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling(TUNEL) assay and western blot for the expressions of caspase-3, cytochrome c, Bax, and Bcl-2 in the retinas were conducted for the detection of apoptotic retinal cell death. The present results showed that the number of TUNEL-positive cells was increased in the retinas of the diabetic rats, whereas treadmill exercise suppressed this number. The expressions of pro-apoptotic factors caspase-3, cytochrome c, and Bax were enhanced and the expressions of anti-apoptotic factor Bcl-2 was decreased in the retinas of the diabetic rats. In contrast, treadmill exercise suppressed the expressions of caspase-3, cytochrome c, and Bax and increased the expression of Bcl-2. The present study demonstrated that treadmill exercise suppressed diabetes-induced apoptotic neuronal cell death in the retinas. Based on the present results, treadmill exercise may be effective therapeutic strategy for the alleviating complications of diabetes patients.

Purification and Characterization of GTP Cyclohydrolase I from Streptomyces tubercidicus, a Producer of Tubercidin

  • Yoo, Jin-Cheol;Han, Ji-Man;Ko, Ok-Hyun;Bang, Hee-Jae
    • Archives of Pharmacal Research
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    • v.21 no.6
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    • pp.692-697
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    • 1998
  • GTP cyclohydrolase I catalyzing the first reaction in the biosynthesis of pterin moiety of folic acid in bacteria, was purified from Streptomyces tubercidicus by at least 203-fold with a yield of 32% to apparent homogeneity, using ammonium sulfate fractionation, DEAE-cellulose, Sepharose CL-6B, and hydroxylapatite column chromatography. The molecular weight of the native enzyme was estimated to be 230,000 daltons by gel permeation chromatography. The purified enzyme gave a single band on sodium dodesyl sulfate-polyacrylamide gel electrophoresis and its molecular weight was apparently 58,000 daltons. These results indicate that the enzyme consists of four subunits with the same molecular weight. The $K_m$ and $V_{max}$ values for GTP of the purified enzyme were determined to be 80${\mu}$M and 90nmol/min (mg protein), respectively. The optimum pH and temperature for the enzyme reaction were pH 7.5-8.5 and $40-42^{\circ}C$, respectively. Coenzyme or metal ion was not required for the enzyme activity. The enzyme activity was inhibited by most divalent cations, while it was slightly activated by potassium ion. In case of nucleotides, CTP, GMP, GDP, and UTP inhibited enzyme activity, among which GDP exhibited the strongest inhibitory effect.

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Inhibition of Cell Growth and Induction of Apoptosis by Euonymus Alatus (Thunb.) Sieb in Human Leiomyomal Smooth Muscle Cells

  • Kim Yi-Geun;Han Ji-Young;Park Young-Soo;Kim Dong-Il;Lee Tae-Kyun
    • The Journal of Korean Medicine
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    • v.25 no.4
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    • pp.108-120
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    • 2004
  • Objective : Euonymus alatus (Thunb.) Sieb (EA) is a traditional Korean herbal medicine, commonly used to treat tumors in Korea and China for centuries. Several earlier studies have indicated that EA exhibits anti-tumor properties, but its mechanism remains to be elucidated. In this study, we evaluated the molecular mechanism of EA in a human uterine leiomyomal smooth muscle cell (ULSMC) line. Methods : This study was evaluated by: (a), morphological changes by using acridine orange/ethidium bromide staining; (b), DNA fragmentation by TdT-mediated dUTP nick end labeling (TUNEL); and (c), sub-G1 cell analysis. Results : This study observed that EA treatment caused apoptotic cell death and depletion of intracellular glutathione (GSH) and that reduction of mitochondrial membrane potential was found to be involved in the initiation of apoptosis by EA. Conclusion : This results show that EA exerted clear cytotoxic effects and strongly inhibited the proliferation of ULSMC.

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Development and Implementation of the Emergency Call Program for a Welfare for the Elderly (노인 복지를 위한 응급 상황 호출 프로그램의 개발 및 구현)

  • Kim, Jung-Hwan;Cho, Myeon-Gyun;Kim, Shik
    • IEMEK Journal of Embedded Systems and Applications
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    • v.8 no.2
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    • pp.79-85
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    • 2013
  • This paper proposes a system that utilizes USN(Ubiquitous Sensor Network), Bluetooth and smart phone to improve the function of senior houses. In typical approach, a system in a senior house either directly accesses the status of elderly people by its sensor or is alerted by elderly people who trigger an emergency bell, derive a decision and take an appropriate action. In addition, it is possible for a designated social worker to check the status of senior patients through monitoring system connected by UTP(Unshielded Uwisted Pair) cables, but the responsible person has to be present to monitor patients' status. However, the new system, suggested in this paper, embed Bluetooth function in a blood pressure gauge, thus the smart phone receives patients' health information such as blood pressure through Bluebooth, if any abnormal event occurs. Consequently, the smart phone sends SMS(Short Message Service) to a responsible social worker or a designated hospital. When this program in the paper becomes a reality, an unmanned system that is able to determine suitable actions for certain events will be established, even if a social worker were absence.

Mechanical properties of blended cements at elevated temperatures predicted using a fuzzy logic model

  • Beycioglu, Ahmet;Gultekin, Adil;Aruntas, Huseyin Yilmaz;Gencel, Osman;Dobiszewska, Magdalena;Brostow, Witold
    • Computers and Concrete
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    • v.20 no.2
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    • pp.247-255
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    • 2017
  • This study aimed to develop a Rule Based Mamdani Type Fuzzy Logic (RBMFL) model to predict the flexural strengths and compressive strengths of blended cements under elevated temperatures. Clinoptilolite was used as cement substitution material in the experimental stage. Substitution ratios in the cement mortar mix designs were selected as 0% (reference), 5%, 10%, 15% and 20%. The data used in the modeling process were obtained experimentally, after mortar specimens having reached the age of 90 days and exposed to $300^{\circ}C$, $400^{\circ}C$, $500^{\circ}C$ temperatures for 3 hours. In the RBMFL model, temperature ($C^{\circ}$) and substitution ratio of clinoptilolite (%) were inputs while the compressive strengths and flexural strengths of mortars were outputs. Results were compared by using some statistical methods. Statistical comparison results showed that rule based Mamdani type fuzzy logic can be an alternative approach for the evaluation of the mechanical properties of concrete under elevated temperature.

Morphological Changes of Radiation-Induced Atretic Follicles in Mouse Ovary

  • Kim Sung-In;Kwon O-Yu;You Kwan-Hee
    • Biomedical Science Letters
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    • v.12 no.2
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    • pp.105-112
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    • 2006
  • This study was undertaken to investigate the morphological changes between normal and atretic follicle after gamma irradiation and treatment of follicle stimulating hormone (FSH). The ovaries of each group of treated immature mice were prepared the paraffin sections after 0, 6, 12, and 24 hours (hrs) of those treatment. Hematoxylin-eosin (HE) stain, reticulin stain, and terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end labeling (TUNEL) immunohistochemical stain were performed on the each paraffin sections. As the results of HE staining, the condensed nuclei of oocytes were observed in the atretic primordial follicles, on the other hand the condensations of granulosa cell nuclei were prominent in the atretic primary, preantral, and antral follicles. Only the granulosa cells of atretic follicle were stained specifically with TUNEL staining but not stained in the theca cells, which suggested granulosa cells degenerated through apoptosis. In the reticulin staining, the basement membranes of atretic follicle which was stained weakly showed irregular structure and detachment from the follicles. The ratio of normal to atretic follicle in control and FSH treated group was about 33% but this ratio increased rapidly over 90% in the 6, 12, and 24 hrs group after the irradiation. It could be suggested that the gamma irradiation is the useful tool far the induction of follicle atresia and immunohistochemical staining methods are essential in the study of follicle atresia.

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The Change of Mitochondrial Fusion and Fission in human Skeletal Muscle with Aging

  • Cho, Hyung-Jun;Park, Soo-Yeon
    • Biomedical Science Letters
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    • v.18 no.2
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    • pp.112-122
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    • 2012
  • A gradual change of molecules that are related in fission and fusion is occurred during aging process. Although aging effects on mitochondrial fusion and fission are investigated, it is still unclear that the extent of the change in mitochondria fusion and fission periodically. In this study we investigated the changes of mitochondrial proteins involved in fusion (Mfn2, Opa1) and fission (Drp1, Fis1) in the human gracilis muscle ranging from 10 to 50 years of age (n=40). The gracilis muscle showed a significant increase in muscle apoptotic changes in the age of 50s compared with 10s by using in situ terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL). The expression levels of Drp1 and Fis1 (P<0.01, P<0.05) mRNA were significantly elevated and the Mfn2 and Opa1 (P<0.01, P<0.05) levels were decreased from older individuals. The ratio of fission and fusion was altered and the level of increment of fission gene was greater than fusion gene decrement in the age of 50s. These findings suggest that changes of mitochondrial fusion and fission proteins related with aging might contribute to aged muscle apoptosis.

Effect of Chronic Inhibition of Nitric Oxide on Blood Pressure and Apoptosis in the Blood Pressure-Associated with Organs

  • Bae, Hyung-Joon
    • Biomedical Science Letters
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    • v.16 no.1
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    • pp.25-32
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    • 2010
  • Sprague-Dawley(SD) rats were orally administered with $N^G$-nitro-L-arginine methyl ester(L-NAME) which inhibits or blocks the production of nitric oxide from L-arginine in vascular endothelial cells and vessel tissue to statistically examine the effects of nitric oxide on some physiological changes such as blood pressure and heart rate, and to confirm the apoptosis induced by the suppressed nitric oxide activity in some related organs under light microscope. Systolic blood pressure significantly increased 28.5% by the chronic treatment of L-NAME for 8 weeks (P<0.001), no significant difference, however, was observed in heart rate between the control group and the L-NAME-treated group regardless of their age. Hematoxylin-eosin staining showed some histological alterations only in kidney among the examined organs; heart, liver, pancreas, and adrenal gland from the L-NAME-treated group. TUNEL (terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling) test showed a strong positive reaction, representing that the chronic treatment of L-NAME facilitates apoptosis, in the cortex and medulla of kidney, but not any significance detectable in the other organs. These results conclude that chronic treatment of L-NAME significantly increases blood pressure, and that the followed inhibition of nitric oxide synthesis occurs a typical inducement of apoptosis in kidney.

Ginseng radix Suppresses Ischemia-induced Increase in c-Fos Expression and Apoptosis in the Hippocampal CA1 Region in Gerbils

  • Park, Keung-Ryol;Jang, Mi-Hyeon;Kim, Chang-Ju;Lee, Choong-Yeol
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.19 no.1
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    • pp.224-229
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    • 2005
  • Ginseng radix, the root of Panax ginseng C.A.Meyer (Araliaceae), has traditionally been used for the treatment of various disorders including cerebrovascular accident (CVA). In the present study, the effect of Ginseng radix on c-Fos expression and apoptosis in the hippocampal CA1 region of gerbils following transient global ischemia was investigated via immunohistochemistry for c-Fos and caspase-3 and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay. Enhanced c-Fos-, TUNEL-, and caspase-3-positivities were detected in the hippocampal CA1 region in ischemic gerbils. Administration of the aqueous extract of Ginseng radix suppressed this ischemia-induced increment in the numbers of c-Fos-, TUNEL-, and caspase-3-positive cells. These results suggest that Ginseng radix has an inhibitive effect on the induction of c-Fos expression and apoptosis seen following transient global ischemia.