• Title/Summary/Keyword: Cytidine

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Nucleotide Sequence on Upstream of the cdd Locus in Bacillus subtilis

  • JONG-GUK KIM;KIM, KYE-WON;SEON-KAP HWANG;JOO-WON SUH;BANG-HO SONG;SOON-DUCK HONG
    • Journal of Microbiology and Biotechnology
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    • v.5 no.3
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    • pp.125-131
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    • 1995
  • A 3, 346 bp of the cdd upstream region in Bacillus subtilis was sequenced from the pSO1 (Song BH and J Neuhard. 1989. Mol. Gen. Genet 216: 462-468) and sequence homology was searched to the known genes in Genbank and European Molecular Biology Laboratory databanks. Five complete and one truncated putative coding sequences deduced from the nucleotide sequence were found through the ORF searching by Genetyx and Macvector software, and one of them was identified as the dgk (diacylglycerol kinase) gene and another, a truncated one, as the phoH (phosphate starvation-inducible gene) gene. The B. subtilis dgk gene, having a role for response to several environmental stress signals, revealed an open reading frame of 134 amino acids with 43.1% of sequence identity to the Streptococcus mutans dgk gene. The carboxy terminal 59 residues of the truncated phoH gene showed 52.7% and 34.5% of sequence identity in amino acids with the corresponding genes of Mycobacterium leprae and Escherichia coli. The four remaining coding sequences consisting of 115, 421, 91, and 91 residues were thought to be unknown ORFs because they have no significant similarity to known genes.

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Phospholipids from Bombycis corpus and their neurotrophic effects

  • Jung, I-Yeon;Kwon, Hak-Cheol;Nam, Sung-Hee;Chang, Seung-Jong;Cho, Sae-Yeon;Hur, In-Young;Kim, Sun-Yeon;Lee, Kang-Ro
    • Proceedings of the Korean Society of Sericultural Science Conference
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    • 2003.04a
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    • pp.46-46
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    • 2003
  • Three phospholiplds (4 6) and three aromatic amines (1 3) were obtained from the methanol extract of Bombycis corpus. Based on spectral data, their structures have been elucidated as nicotiamide(1), cytidine(2), adenine(3), 1-O-(9Z-octadecenoyl)-2-O-(8Z, llZ-octadecadienoyl)-sn-glycero-3-phosphorylcholine(4), 1,2-di-O-hexadecanoly-sn-glycero-3-phosphorylcholine(5) and 1,2-di-O-9Z-octadecenoly-sn-glycero-3-phosphorylcholine(6). (omitted)

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Purification and Properties of Intracellular Cytosine Deaminase from Chromobacterium violaceum YK 391

  • KIM , JUNG;YU, TAE-SHICK
    • Journal of Microbiology and Biotechnology
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    • v.14 no.6
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    • pp.1182-1189
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    • 2004
  • Cytosine deaminase (cytosine aminohydrolase, EC 3.5.4.1) stoichiometrically catalyzes the hydrolytic deamination of cytosine and 5-fluorocytosine to uracil and 5-fluorouracil, respectively. The intracellular cytosine deaminase from Chromobacterium violaceum YK 391 was purified to apparent homogeneity with 272.9-fold purification with an overall yield of $13.8\%$. The enzyme consisted of dimeric polypeptides of 63 kDa, and the total molecular mass was calculated to be approximately 126 kDa. Besides cytosine, the enzyme deaminated 5-fluorocytosine, cytidine, 6-azacytosine, and 5-methylcytosine, but not 5-azacytosine. Optimum pH and temperature for the enzyme reaction were 7.5 and $30^{\circ}C$, respectively. The enzyme was stable at pH 6.0 to 8.0, and at 30T for a week. About $70\%$ of the enzyme activity was retained at $60^{\circ}C$ for 5 min. The apparent $K_{m}$ values for cytosine, 5-fluorocytosine, and 5-methylcytosine were calculated to be 0.38 mM, 0.87 mM, and 2.32 mM, respectively. The enzyme activity was strongly inhibited by 1 mM $Hg^{2+},\;Zn^{2+},\;Cu^{2+},\;Pb^{2+},\;and\;Fe^{3+}$, and by o-phenanthroline, $\alpha,\;{\alpha}'$-dipyridyl, p-choromercuribenzoate, N-bromosuccinimide, and cWoramine­T. In addition, the enzyme activity was strongly inhibited by I mM 2-thiouracil, and weakly inhibited by 2-thiocytosine, or 5-azacytosine. Finally, intracellular and extracellular cytosine deaminases from Chromobacterium violaceum YK 391 were found to have a different optimum temperature, apparent $K_{m}$ value, and molecular mass.

A Literature Study on Acupuncture for Cerebral Palsy -Based on the Current Traditional Chinese Medical Journals- (뇌성마비의 침자(鍼刺) 치료에 대한 문헌적 고찰 -최근 중의잡지를 중심으로-)

  • Ha, Su-Yun;Min, Sang-Yeon;Kim, Jang-Hyun
    • The Journal of Pediatrics of Korean Medicine
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    • v.23 no.1
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    • pp.205-228
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    • 2009
  • Objectives The purpose of this study is to investigate the current acupuncture therapy of cerebral palsy. Methods We investigated the Chinese clinical papers which were published in the last 10 years(from 1999 to 2008). We found these papers from the oriental medical library in university and we also used the China National Knowledge Infrastructure(CNKI) through the internet and selected 32 papers for analyzing. Results Most papers were described the effect of acupuncture or acupoint-injection. This is more effective way to treat than the general rehabilitation treatment such as the physical therapy, the occupational therapy, and the speech therapy. Acupuncture or acupoint-injection has overall $80{\sim}100%$ of rehabilitation rate. The younger the children were, the longer the treatment period was, and the more successful in treatment. The acupuncture was often used with the general acupuncture and scalp acupuncture. Commonly used major acupuncture points were sishencong(四神聰), bohui(百會), zusanli(足三里), yundongqu(運動區), pinghengqu(平衡區), quchi(曲池), and sanyinjiao(三陰交). Commonly used main meridian pathways were bladder, governor vessel, gallbladder, large intestine, stomach, small intestine meridian. Head is the common site for acupuncture. The main acupoint-injection points were zusanli(足三里), dazhui(大椎), shenshu(腎兪), yamen(啞門), neiguan(內關), and fengchi(風池). For the injection, brain activator, ganglioside M1, cerebroprotein hydrolysate, cytidine diphosphate choline, Vit B1, Vit B12, the salviae root, the safflower were commonly used. Conclusions Acupuncture and acupoint-injection have been shown as an effective treatment on cerebral palsy. The acupuncture was used often the general acupuncture and scalp acupuncture all together. Commonly used main acupuncture points were sishencong, bohui, zusanli, yundongqu, pinghengqu, quchi, sanyinjiao.

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Euchromatin histone methyltransferase II (EHMT2) regulates the expression of ras-related GTP binding C (RRAGC) protein

  • Hwang, Supyong;Kim, Soyoung;Kim, Kyungkon;Yeom, Jeonghun;Park, Sojung;Kim, Inki
    • BMB Reports
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    • v.53 no.11
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    • pp.576-581
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    • 2020
  • Dimethylation of the histone H3 protein at lysine residue 9 (H3K9) is mediated by euchromatin histone methyltransferase II (EHMT2) and results in transcriptional repression of target genes. Recently, chemical inhibition of EHMT2 was shown to induce various physiological outcomes, including endoplasmic reticulum stress-associated genes transcription in cancer cells. To identify genes that are transcriptionally repressed by EHMT2 during apoptosis, and cell stress responses, we screened genes that are upregulated by BIX-01294, a chemical inhibitor of EHMT2. RNA sequencing analyses revealed 77 genes that were upregulated by BIX-01294 in all four hepatic cell carcinoma (HCC) cell lines. These included genes that have been implicated in apoptosis, the unfolded protein response (UPR), and others. Among these genes, the one encoding the stress-response protein Ras-related GTPase C (RRAGC) was upregulated in all BIX-01294-treated HCC cell lines. We confirmed the regulatory roles of EHMT2 in RRAGC expression in HCC cell lines using proteomic analyses, chromatin immune precipitation (ChIP) assay, and small guide RNA-mediated loss-of-function experiments. Upregulation of RRAGC was limited by the reactive oxygen species (ROS) scavenger N-acetyl cysteine (NAC), suggesting that ROS are involved in EHMT2-mediated transcriptional regulation of stress-response genes in HCC cells. Finally, combined treatment of cells with BIX-01294 and 5-Aza-cytidine induced greater upregulation of RRAGC protein expression. These findings suggest that EHMT2 suppresses expression of the RRAGC gene in a ROS-dependent manner and imply that EHMT2 is a key regulator of stress-responsive gene expression in liver cancer cells.

Phospholipase Activities in Clinical and Environmental Isolates of Acanthamoeba

  • Matin, Abdul;Jung, Suk-Yul
    • Parasites, Hosts and Diseases
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    • v.49 no.1
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    • pp.1-8
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    • 2011
  • The pathogenesis and pathophysiology of Acanthamoeba infections remain incompletely understood. Phospholipases are known to cleave phospholipids, suggesting their possible involvement in the host cell plasma membrane disruption leading to host cell penetration and lysis. The aims of the present study were to determine phospholipase activities in Acanthamoeba and to determine their roles in the pathogenesis of Acanthamoeba. Using an encephalitis isolate (T1 genotype), a keratitis isolate (T4 genotype), and an environmental isolate (T7 genotype), we demonstrated that Acanthamoeba exhibited phospholipase $A_2$ (PLA$_2$). and phospholipase D (PLD) activities in a spectrophotometry-based assay. Interestingly, the encephalitis isolates of Acanthamoeba exhibited higher phospholipase activities as compared with the keratitis isolates, but the environmental isolates exhibited the highest phospholipase activities. Moreover, Acanthamoeba isolates exhibited higher PLD activities compared with the PLA$_2$. Acanthamoeba exhibited optimal phospholipase activities at $37^{\circ}C$ and at neutral pH indicating their physiological relevance. The functional role of phospholipases was determined by in vitro assays using human brain microvascular endothelial cells (HBMEC), which constitute the blood-brain barrier. We observed that a PLD-specific inhibitor, i.e., compound 48/80, partially inhibited Acanthamoeba encephalitis isolate cytotoxicity of the host cells, while PLA$_2$-specific inhibitor, i.e., cytidine 5'-diphosphocholine, had no effect on parasite-mediated HBMEC cytotoxicity. Overall, the T7 exhibited higher phospholipase activities as compared to the T4. In contract, the T7 exhibited minimal binding to, or cytotoxicity of, HBMEC.

Effect of $1-{\beta}-D-Arabinofuranosylcytosine$ on the Cytoplasmic Organelles of the Hepatocytes in Albino Mice ($1-{\beta}-D-Arabinofuranosylcytosine$이 Mouse의 간세포소기관(肝細胞小器官)에 미치는 영향(影響))

  • Kim, S.Y.;Lee, K.S.
    • Applied Microscopy
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    • v.13 no.1
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    • pp.13-30
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    • 1983
  • [ $1-{\beta}-D-Arabinofuranosylcytosine$ ](ara-C), which is a pyrimidine nucleoside analog is cytotonic to mammalian cells in culture and is active in vitro and in vivo against a variety of DNA viruses. The precise mechanism of action of ara-C has not been determined, although ara-C is thought to act as an antimetabolite, interfering with the synthesis of deoxyribonucleic acid(DNA). Cytosine arabinoside originally seemed to act principally by inhibiting the conversion of cytidine to deoxytidine, thus inhibiting DNA synthesis. But recent data suggest that effects upon DNA polymerase and effects via incorporation into DNA and RNA may well be of equal importance. The author have demonstrated the effect of cytosine arabinoside on the hepatocytes of albino mice treated with ara-C, observing changes in the cytoplasmic organelles of the hepatocytes. A total of 120 healthy male albino mice were divided into the control and ara-C treated groups. The animals of the ara-C group were given 10mg. per kg of body weight of mouse ara-C in physiological saline solution and the animals of control group were given physiological saline solution, intraperitoneally. After an administration of ara-C or physiological saline solution, the animal were killed at. interval of 6, 12, and 24 hours. The specimens, which were obtained from the left anterier lobe of the liver, were stained with uranyl acetate and lead citrate and observed with JEM 100B electron microscope. The results were obtained as follow: A pronounced dilatation, sacculation and fragmentation of the cisterane of rough endoplasmic reticulum with dissociation of membrane bound-ribosomes, disaggregation of free ribosomes in the cytoplasm, proliferation of the smooth endoplasmic reticulum associated with depletion of glycogen paracles, atrophies of Golgi complex, production of numerous lipid droplets, and formation of antophagic vacuoles, multivesicular bodies and residual bodies are recognized in the hepatocytes of ara-C treated mice. Consequently it is suggested that cytosine arabinoside would induce a changes of the cytoplasmic organelles of the hepatocytes in albino mice.

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Phospholipids from Bombycis corpus and Their Neurotrophic Effects

  • Kwon, Hak-Cheol;Jung, I-Yeon;Cho, Se-Yeon;Cho, Ock-Ryun;Yang, Min-Cheol;Lee, Sung-Ok;Hur, Jin-Young;Kim, Sun-Yeou;Yang, Jong-Beom;Lee, Kang-Ro
    • Archives of Pharmacal Research
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    • v.26 no.6
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    • pp.471-477
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    • 2003
  • Three phospholipids (4-6) and three aromatic amines (1-3) were obtained from the methanol extract of Bombycis corpus. Based on spectral data, their structures have been elucidated as nicotiamide (1), cytidine (2), adenine (3), 1-Ο-(9Z-octadecenoyl)-2-Ο-(8Z,11Z-octadecadienoyl)-sn-glycero-3-phosphorylcholine (4), 1,2-di-Ο-hexadecanoyl-sn-glycero-3-phosphorylcholine (5) and 1,2-di-Ο-9Z-octadecenoyl-sn-glycero-3-phosphorylcholine (6). We examined the effects of compounds on synthesis of NGF in cultured astrocytes. By RT-PCR analysis, expresison of NGF mRNA in astrocytes cultured in serum-starvation increased after the addition of phospholipid (10 $\mu$M). The NGF content in the culture medium was significantly increased by compound 5, compared with the control value. These results suggest that three phospholipid compounds isolated from the methanol extract of Bombycis corpus may exert neurotrophic effects by stimulation of NGF synthesis in astrocytes.

Cloning of hnRNP E1 cDNA via yeast two-hybrid system and a study on protein-protein interaction between hnRNP E1 and hnRNP K (이스트 two-hybrid 시스템을 이용한 hnRNP E1 cDNA의 클로닝과 hnRNP E1-hnRNP K 상호결합에 대한 연구)

  • Choi, Mie-Young
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.9 no.6
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    • pp.1795-1799
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    • 2008
  • The heterogeneous nuclear ribonucleoprotein K (hnRNP K) is a component of hnRNP complexes. This protein binds strongly to cytidine-rich RNA/DNA sequences. It is a nucleocytoplasmic shuttling protein. To investigate the functions of hnRNP K, I searched for hnRNP K-interacting proteins in HeLa cDNA library using a yeast two-hybrid screening system. One of the cDNA clones is identical to human hnRNP E1 (poly(rC) binding protein 1) cDNA (GenBank accession number XM_031585). In this study, hnRNP K is shown to specifically interact with hnRNP E1 in yeast two-hybrid system and in vitro biochemical assay.

Nucleotide Analysis in Korean Dairy Products Using High- Performance Liquid Chromatography with Diode Array Detector

  • Won, Jong-Eun;Bang, Han-Yeol;Kwak, Byung-Man;Park, Jong-Su;Kim, Gui-Ran;Kwon, Joong-Ho
    • Food Science of Animal Resources
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    • v.39 no.1
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    • pp.93-101
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    • 2019
  • Nucleotides play important roles in numerous intracellular biochemical processes and are used in infant formulas and other dairy products. However, domestic analytical methods for assessing nucleotide content in products have not yet been established, and therefore, methods for determining nucleotide content are urgently required. A rapid and simple analytical method for determining the content of five types of nucleotides in dairy products was improved using solid phase extraction clean-up and high-performance liquid chromatography with diode array detector. The extraction solvent used in the AOAC method was not well dissolved and was changed to hydrophilic EDTA-Na. In addition, the results obtained using the isocratic elution method and a single wavelength were similar to those obtained using the AOAC method, and the time taken for analysis was shortened from 40 min to 25 min. The process of method validation revealed the following parameters: accuracy (84.69%-102.72%), precision (1.51%-6.82%), linearity (0.999), and limit of detection (cytidine 5'-monophosphate, 0.09 mg/L; uridine 5'-monophosphate, 0.11 mg/L; adenosine 5'-monophosphate, 0.12 mg/L; guanosine 5'-monophosphate, 0.11 mg/L; and inosine 5'-monophosphate, 0.14 mg/L). The method was also used to determine the nucleotide concentration in 25 samples (infant formulas, 1.99-29.39 mg/100 g; and cow milk, 0.28-0.83 mg/100 g). The newly improved method was appropriate for analyzing nucleotides in infant formulas and other dairy products faster when compared to conventional methods.