• Title/Summary/Keyword: r-GTP.

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Hypermethylation and Clinicopathological Significance of RASAL1 Gene in Gastric Cancer

  • Chen, Hong;Pan, Ying;Cheng, Zheng-Yuan;Wang, Zhi;Liu, Yang;Zhao, Zhu-Jiang;Fan, Hong
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.11
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    • pp.6261-6265
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    • 2013
  • Background: Recent studies have suggested that expression of the RAS protein activator like-1 gene (RASAL1) is decreased in gastric carcinoma tissues and cell lines, indicated a role in tumorigenesis and development of gastric cancer. Reduced expression of RASAL1 could result in aberrant increase of activity of RAS signaling pathways in cancer cells. However, the exact mechanism which induces down-regulation of the RASAL1 gene remains unclear. This study aimed to determine the methylation status and regulation of RASAL1 in gastric cancer. Materials and Methods: Using the methylation-specific polymerase chain reaction (MSP), the methylation status of CpG islands in the RASAL1 promoter in gastric cancers and paired adjacent non-cancerous tissues from 40 patients was assessed and its clinicopathological significance was analyzed. The methylation status of RASAL1 in gastric cancer lines MKN-28, SGC-790l, BGC-823, as well as in normal gastric epithelial cell line GES-l was also determined after treatment with a DNA methyltransferase inhibitor, 5-aza-2'-doexycytidine (5-Aza-CdR). RAS activity (GAS-GTP) was assessed through a pull-down method, while protein levels of ERK1/2, a downstream molecule of RAS signaling pathways, were determined by Western blotting. Results: The frequencies of RASAL1 promoter methylation in gastric cancer and paired adjacent non-cancerous tissues were 70% (28/40) and 30% (12/40) respectively (P<0.05). There were significantly correlations between RASAL1 promoter methylation with tumor differentiation, tumor size, invasive depth and lymph node metastasis in patients with gastric cancer (all P<0.05), but no correlation was found for age or gender. Promoter hypermethylation of the RASAL1 gene was detected in MKN-28, SGC-790l and BGC-823 cancer cells, but not in the normal gastric epithelial cell line GES-1. Elevated expression of the RASAL1 protein, a decreased RAS-GTP and p-ERK1/2 protein were detected in three gastric cancer cell lines after treatment with 5-Aza-CdR. Conclusions: Aberrant hypermethylation of the RASAL1 gene promoter frequently occurs in gastric cancer tissues and cells. In addition, the demethylating agent 5-Aza-CdR can reverse the hypermethylation of RASAL1 gene and up-regulate the expression of RASAL1 significantly in gastric cancer cells in vivo. Our study suggests that RASAL1 promoter methylation may have a certain relationship with the reduced RASAL1 expression in gastric cancer.

Identification of Three Positive Regulators in the Geldanamycin PKS Gene Cluster of Streptomyces hygroscopicus JCM4427

  • Kim, Won-Cheol;Lee, Jung-Joon;Paik, Sang-Gi;Hong, Young-Soo
    • Journal of Microbiology and Biotechnology
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    • v.20 no.11
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    • pp.1484-1490
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    • 2010
  • In the Streptomyces hygroscopicus JCM4427 geldanamycin biosynthetic gene cluster, five putative regulatory genes were identified by protein homology searching. Among those genes, gel14, gel17, and gel19 are located downstream of polyketide synthase genes. Gel14 and Gel17 are members of the LAL family of transcriptional regulators, including an ATP/GTP-binding domain at the N-terminus and a DNA-binding helix-turn-helix domain at the C-terminus. Gel19 is a member of the TetR family of transcriptional regulators, which generally act to repress transcription. To verify the biological significance of the putative regulators in geldanamycin production, they were individually characterized by gene disruption, genetic complementation, and transcriptional analyses. All three genes were confirmed as positive regulators of geldanamycin production. Specifically, Gel17 and Gel19 are required for gel14 as well as gelA gene expression.

Regulation of c-fos promoter through interaction between dopamine D3 receptor and RGL, ral GDP dissociation stimulator-like

  • Park, Ju-Ran;Kim, So-Young;Kim, Kyeong-Man
    • Proceedings of the PSK Conference
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    • 2003.10b
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    • pp.89.2-89.2
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    • 2003
  • Ral GDP dissociation stimulator (Ral GDS) has been found to be an effector protein of Ras, and Ral, a member of small GTP-binding protein (G protein) superfamily, has been suggested to act downstream of Ras. Ral GDP dissociation stimulator-like (RGL) shares 50% amino acid identity with Ral GDP dissociation stimulator, and assumed to possess similar functional role. Using yeast two-hybrid screening, we found that dopamine D3 receptor interacts with RGL. Since RGL is known to regulate the expression of c-fos promoter, effects of D3R on gene expression of c-fos promoter was studied. (omitted)

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Hepatotoxicity in treatment of canine dermatophytosis with ketoconazole (피부사상균 감염개에서 Ketoconazole 경구투여시의 간독성에 관한 연구)

  • Bae, Seong-su;Kim, Cheol-ho;Kim, Tae-yung;Kang, Chung-boo
    • Korean Journal of Veterinary Research
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    • v.45 no.2
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    • pp.255-261
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    • 2005
  • The purpose of this study is to compare hepatotoxicity of each treatment for dermatophytosis; one is the administration of the ketoconazole only and the other, ketoconazole with diphenyl-dimeththyl-dicarboxylate. Have chosen the range of 14-24 months of healthy dogs divided by two groups (group 1 and group 2) for the experiment of which test proved positive in dermatophytosis diagnosis and showed normal reaction in terms of physical examination, blood chemistry and especially of liver function. Group 1 was administrated ketoconazole orally at 10 mg/kg/day and of same dose of ketoconazole with diphenyl-dimethyl-dicarboxylate for group 2. After administering, we have tested two groups by blood collecting every one week in order to check the differences of hepatotoxicity state through AST, ALT and r-GTP, the barometers of liver function which lasted for 12 weeks. Moreover, tested Indocyanine Green (ICG), known as susceptible gauge of function of excretion before starting the experiment and tested ICG as well after 12 weeks. The experiment of result the value of group 1 in AST, ALT and r-GTP has been highly rised after administering ketoconazole for 10 weeks meanwhile, of group 2 has shown a steady state troughout the whole experiment. For ICG test, we injected 0.5 mg/kg of ICG into a vein for both groups and tested the retention rate at regular interval of 15, 30, 45 minutes. The results of retention rate in two groups were similar to before the drug administration. However, after 12 weeks the retention rate of group 1 has been delayed, on the other hand, retention rate of group 2 were a steady state. In conclusion, the administration of ketoconazole only for a long period of time induced hepatotoxicity where as, the administration of ketoconazole with diphenyl-dimethyl-dicarboxylate didn't induce hepatotoxicity. Therefore, when doctors prescribes for a dog with dermatophytosis should not administrate ketoconnazole itself but with diphenyl-dimethyl-dicarboxylate and one who has abnormal condition of liver function should not be prescribed ketoconazole treatment. If there is a case needed to prescribe ketoconazole treatment, the regular monitoring should be accompanied by at the same time.

Discrepancy between in vitro and in vivo Effect of $G{\alpha}_s$ Gene Mutation on the mRNA Expression of TRH Receptor

  • Park, Seung-Joon;Yang, In-Myung;Yim, Sung-Vin;Chung, Joo-Ho;Jung, Jee-Chang;Ko, Kye-Chang;Kim, Young-Seol;Choi, Young-Kil
    • The Korean Journal of Physiology and Pharmacology
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    • v.2 no.1
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    • pp.101-108
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    • 1998
  • We investigated the effect of ${\alpha}-subunit$ of the stimulatory GTP-binding protein ($G{\alpha}_s$) gene mutation on the expression of the thyrotropin-releasing hormone (TRH) receptor (TRH-R) gene in GH3 cells and in growth hormone (GH)-secreting adenomas of acromegalic patients. In the presence of cyclohexicmide, forskolin and isobutylmethylxanthine, cholera toxin, and GH-releasing hormone (GHRH) decreased rat TRH-R (rTRH-R) gene expression by about 39%, 43.7%, and 46.7%, respectively. Transient expression of a vector expressing mutant-type $G{\alpha}_s$ decreased the rTRH-R gene expression by about 50% at 24 h of transfection, whereas a wild-type $G{\alpha}_s$ expression vector did not. The transcript of human TRH-R (hTRH-R) gene was detected in 6 of 8 (75%) tumors. Three of them (50%) showed the paradoxical GH response to TRH and the other three patients did not show the response. The relative expression of hTRH-R mRNA in the tumors from patients with the paradoxical response of GH to TRH did not differ from that in the tumors from patients without the paradoxical response. Direct PCR sequencing of $G{\alpha}_s$ gene disclosed a mutant allele and a normal allele only at codon 201 in 4 of 8 tumors. The paradoxical response to TRH was observed in 2 of 4 patients without the mutation, and 2 of 4 patients with the mutation. The hTRH-R gene expression of pituitaty adenomsa did not differ between the tumors without the mutation and those with mutation. The present study suggests that the expression of TRH-R gene is not likely to be a main determinant for the paradoxical response of GH to TRH, and that $G{\alpha}_s$ mutation may suppress the gene expression of TRH-R in GH-secreting adenoma. However, a certain predisposing factor(s) may play an important role in determining the expression of TRH-R.

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Non-ribosomal Ribosome Assembly Factors in Escherichia coli (Escherichia coli 에서 리보솜 조립과정에 관여하는 단백질들)

  • Choi, Eunsil;Hwang, Jihwan
    • Journal of Life Science
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    • v.24 no.8
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    • pp.915-926
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    • 2014
  • The ribosome is a protein synthesizing machinery and a ribonucleoprotein complex that consists of three ribosomal RNAs (23S, 16S and 5S) and 54 ribosomal proteins in bacteria. In the course of ribosome assembly, ribosomal proteins (r-protein) and rRNAs are modified, the r-proteins bind to rRNAs to form ribonucleoprotein complexes which are folded into mature ribosomal subunits. In this process, a number of non-ribosomal trans-acting factors organize the assembly process of the components. Those factors include GTP- and ATP-binding proteins, rRNA and r-protein modification enzymes, chaperones, and RNA helicases. During ribosome biogenesis, they participate in the modifications of ribosomal proteins and RNAs, and the assemblies of ribosomal proteins with rRNAs. Ribosomes can be assembled from a discrete set of components in vitro, and it is notable that in vivo ribosome assembly is much faster than in vitro ribosome assembly. This suggests that non-ribosomal ribosome assembly factors help to overcome several kinetic traps in ribosome biogenesis process. In spite of accumulation of genetic, structural, and biochemical data, not only the entire procedure of bacterial ribosome synthesis but also most of roles of ribosome assembly factors remain elusive. Here, we review ribosome assembly factors involved in the ribosome maturation of Escherichia coli, and summarize the contributions of several ribosome assembly factors which associate with 50S and 30S ribosomal subunits, respectively.

Study of 25-Hydroxyvitamin D3 concentration according to the Hepatitis B virus (Hepatitis B virus에 따른 25-Hydroxyvitamin D3 농도 영향 연구)

  • Kim, Jean-Soo;Lee, Dong-Yeop
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.14 no.6
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    • pp.2743-2748
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    • 2013
  • The vitamin gets through food and the sun. But there is case that density alters by certain factor in vitamin. So this research analyzed effect of the liver function factor and change of $25OH-VIT.D_3$ density. Research target is a person who undergo health medical examination in the certain university hospital medical examination center from August, 2012 to December, 2012. Comparison classified to experiment group and control group. As a result, variable that influence in $25OH-VIT.D_3$ density showed by age and B type inflammation of the liver titer. Specially, B type inflammation of the liver titer displayed reverse corelation ($R^2$=0.40). Therefore, inflammation of the liver carrier must manage liver fuction test and density of $25OH-VIT.D_3$.

Hematological and blood chemical changes of rabbits infected with viral hemorrhagic disease virus (토끼 출혈성 바이러스에 감염된 토끼의 혈액상과 혈액화학치의 변화)

  • Yoon, In-joong;Jeon, Yun-seong
    • Korean Journal of Veterinary Research
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    • v.30 no.1
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    • pp.73-78
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    • 1990
  • Hematological and blood chemical changes of rabbits infected with rabbit hemorrhagic disease virus were studied and the results were summarized as follows: 1. Total leukocyte count ($2,410{\pm}1,076/{\mu}l$), lymphocyte count ($1,582{\pm}632.5{\mu}l$) and heterophil count ($705{\pm}411.1{\mu}l$) were significantly decreased after 24 hours of the infection (p<0.01). However, no significant changes were observed in monocyte, eosinophil and basophil numbers. 2. A significant increase of aspartate aminotransferase (96IU/L), alanine aminotransferase (96IU/L) and alkaline phosphatase ($401.1{\pm}131.8IU/L$) was observed (p<0.01). 3. A moderate increase of BUN ($26.9{\pm}3.6mg/100ml$) and creatinine ($3.2{\pm}1.9mg/100ml$) was observed (p<0.05). 4. No significant changes of r-GTP, thymol turbidity, glucose, cholesterol, albumin, total serum protein, fibrinogen were observed.

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The Clinical Study on Abnormal Liver Function Patients Caused by Obesity (비만증(肥滿症)과 간기능(肝機能) 이상(異常)을 동반(同伴)한 환자(患者) 11례(例)에 대한 임상적(臨床的) 고찰(考察))

  • Lim, Choon-Woo;Kim, Kyung-Hoon;Park, Young-Jun;Park, Jou-Han;Yun, Bo-Hyeon
    • The Journal of Internal Korean Medicine
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    • v.22 no.4
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    • pp.547-555
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    • 2001
  • Objectives: Obesity is regarded as the aggregation of needless risk factors, for instance, cardiovascular disease, joint disease, induce cancer. We studied on interrelation between abnormal liver function and obesity. Methods: We analyzed liver function, T.Cholesterol, Triglyceride before and after lose weight treatment. The collateral condition is over 6 weeks period on obesity treatment, no liver injury and no complicated another disease on personal past history and found out abnormal impression on biochemical liver function blood test. Results: The improvement rate of LFT, compare with before treatment is 10.6% in T.Bilirubin, 11.1% in ALP, 21% in AST, 38% in ALT, 37.3% in r-GTP, 9.2% in LDH and decreased 2.7% in T.Protein, increased 2.3% in Albumin. Hyperlipidemia is 19.4% in T.Cholesterol, 42.5% in Triglyceride. Conclusions: LFT and Hyperlipidemia of abnormal liver function patients, caused by obesity, is improved to normal limit in proportion to reduce patient's weight.

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Studies on the hematological and blood chemical findings for dry and lactating periods (건유(乾乳)와 필유(泌乳)에 따른 유우(乳牛)의 혈액(血液) 및 혈액화학상(血液化學像)의 변화에 관한 연구)

  • Kang, Chung-boo;Lee, Kuk-cheon;Lee, Soon-sun;Cho, Hee-taek;Ahn, Dong-won
    • Korean Journal of Veterinary Research
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    • v.29 no.4
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    • pp.437-443
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    • 1989
  • This experiment was carried out to investigate the hematological and blood chemical values in dairy cattle during the dry and lactating period. Blood was collected from six healthy dairy cattle in Gyeongnam province. Leucocytes count, hemoglobin concentrations and the percentage of packed cell volume were lowest at the early lactation period. Leucocytes count was not affected during dry the to lactating period. The differential count of eosinophilic leucocytes was low at the late lactation period, while the basophilic leucocytes was high at the dry period. Inorganic phosphorus value was below under normal level. Creatinine value was from 1 to 2mg/100ml of serum. Positive reaction to CRP was shown in normal dairy cattle. The values of AST and ALT were higher during the dry than lactating period. r-GTP and total cholesterol tend to be decreased during the lactating period. CPK value was not affected for dry and lactating period.

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