• 제목/요약/키워드: $P2Y_{12}$ receptor

검색결과 252건 처리시간 0.028초

Recombinant Expression, Isotope Labeling and Purification of the Vitamin D Receptor Binding Peptide

  • Chae, Young-Kee;Singarapu, Kiran;Westler, W. Milo;Markley, John L.
    • Bulletin of the Korean Chemical Society
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    • 제32권12호
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    • pp.4337-4340
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    • 2011
  • The vitamin D receptor binding peptide, VDRBP, was overexpressed as a fused form with the ubiquitin molecule in Rosetta(DE3)pLysS, a protein production strain of Escherichia coli harboring an induction controller plasmid. The fusion protein was bound to the immobilized metal ions, and the denaturation and renaturation of the fusion protein were performed as a part of the purification procedure. After the elution of the fusion protein, the peptide hormone was released from its fusion partner by using yeast ubiquitin hydrolase (YUH), and subsequently purified by reverse phase chromatography. The purity of the resulting peptide fragment was checked by MALDI-TOF mass and NMR spectroscopy. The final yields of the target peptide were around 5 and 2 mg per liter of LB and minimal media, respectively. The recombinant expression and purification of this peptide will enable structural and functional studies using multidimensional NMR spectroscopy and X-ray crystallography.

인체유방암 세포주 MCF-7 세포에서 genistein의 Aryl Hydrocarbon Receptor와 Cytochrome P450 1A1에 대한 영향 (Effect of Genistein on the Aryl Hydrocarbon Receptor and Cytochrome P450 1A1 in MCF-7 Human Breast Carcinoma Cells)

  • 한은희;김지영;정혜광
    • Environmental Analysis Health and Toxicology
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    • 제21권1호
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    • pp.13-19
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    • 2006
  • 화학적 예방효과가 있는 식물성 에스트로젠은 다양한 환성을 나타내며 여러 세포 수용체와 상호작용한다. Genistein은 isoflavone의 주요물질 중의 하나로 콩류에 존재하며 대표적인 식물성 에스트로젠이다. 본 논문에서는 유방암 세포주인 MCF-7에서 aryl hydrocarbon receptor(AhR)에 의해 매개되는 발암물질 활성화 경로에 대한 genistein의 영향을 살펴보았다. 세포에 genistein을 처리할 경우 cytochrome P450 1A1(CYP1A1) 약물대사효소의 특이적인 효소반응인 7-ethoxyresorufin O-deethylase (EROD) 활성도와 CYP1A1의 유전자 발현이 genistein의 농도 의존적으로 증가하였다. Genistein과 발암물질인 방향족탄화 수소 7, 12-dimethylbenz[a]anthracene(DMBA)를 동시 처리하였을 경우 DMBA에 의해 유도되어 증가된 EROD활성도와 CYP1A1의 유전자 발현이 genistein에 의해 감소하였다. 랫트의 간에서 분리한 세포질을 이용하여 genistein과 AhR의 대표적인 ligand인 2,3,7,8-tetrachlorodibenzo-p-dioxin과 경쟁적 결합에 대한 영향을 조사한 결과 genistein이 AhR에 경쟁적으로 결합함을 알 수 있었다. 이러한 결과들은 genistein이 천연 AhR ligand임을 암시한다. 따라서, 식물성 에스트로젠인 genistein은 AhR경로의 길항제/항진제로 작용할 수 있을 것으로 사료된다.

가임신 흰쥐 자궁조직 분화에 미치는 난소 스테로이드 호르몬의 영향 (The Effect of the Ovarian Steroid Hormone on the Differenciation of the Pseudopregnanct Rat Uterus)

  • 김성례
    • Clinical and Experimental Reproductive Medicine
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    • 제22권2호
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    • pp.155-161
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    • 1995
  • The present investigation has been undertaken to elucidate the differentiation mechanism the uterus which is the environment of the embryo development, by demonstrating the role of ovarian steroids hormone in the decidualization of the pseudopregnant rat uterus. To determine the effect of ovarine steroids and artificial stimulation (trauma) on the differenciation of the uterine endometrium and decidualization for implantation, attempt was made to measure concentrations of serum estradiol($E_2$), progesterone($P_4$) and nuclear $P_4$ receptor in the traumatized and non-traumatized uterine tissue of the pseudopregnant rat. The results obtained are as followings : The concentration of serum $E_2$ on day 9(implantation stage) was similar in both of intact pseudopregnant rat(47.63pg/ml) and normal pregnant rat(40.71pg/ml). And among the treated groups, $E_2$ concentration was highest in the $E_2$ treated group in comparision with intact control group(relative value; 73.27%). The concentration of serum $P_4$ was also highest in the $P_4$ treated group(23.12pg/ml). Relative value of $P_4$ treated group in comparision with intact group(24.88pg/ml) was 92.93%. The nuclear $P_4$ receptor levels in the artificial traumatized groups were higher compared with the non-traumatized control groups. This study, therefore, clearly demonstrates that the methods for inducing pseudopregnant (vagina tapping;120/min) and inducing decidualization(oil injection; 0.1ml/uterine horn) appear to be effective, $P_4$ appears to be effective in the differenciation of the uterine endometrial tissue for the implantation process. Concentration of serum $P_4$ seems to be well correlated with the level of the nuclear $P_4$ receptor during the early embryo development. These results seem to be well correlated with ALPase activities in the normal and pseudopregnant rat uterus shown in the previous study.

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Dietary Niacin Supplementation Suppressed Hepatic Lipid Accumulation in Rabbits

  • Liu, Lei;Li, Chunyan;Fu, Chunyan;Li, Fuchang
    • Asian-Australasian Journal of Animal Sciences
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    • 제29권12호
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    • pp.1748-1755
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    • 2016
  • An experiment was conducted to investigate the effect of niacin supplementation on hepatic lipid metabolism in rabbits. Rex Rabbits (90 d, n = 32) were allocated to two equal treatment groups: Fed basal diet (control) or fed basal diet with additional 200 mg/kg niacin supplementation (niacin). The results show that niacin significantly increased the levels of plasma adiponectin, hepatic apoprotein B and hepatic leptin receptors mRNA (p<0.05), but significantly decreased the hepatic fatty acid synthase activity and adiponectin receptor 2, insulin receptor and acetyl-CoA carboxylase mRNA levels (p<0.05). Plasma insulin had a decreasing tendency in the niacin treatment group compared with control (p = 0.067). Plasma very low density lipoproteins, leptin levels and the hepatic adiponectin receptor 1 and carnitine palmitoyl transferase 1 genes expression were not significantly altered with niacin addition to the diet (p>0.05). However, niacin treatment significantly inhibited the hepatocytes lipid accumulation compared with the control group (p<0.05). In conclusion, niacin treatment can decrease hepatic fatty acids synthesis, but does not alter fatty acids oxidation and triacylglycerol export. And this whole process attenuates lipid accumulation in liver. Besides, the hormones of insulin, leptin and adiponectin are associated with the regulation of niacin in hepatic lipid metabolism in rabbits.

Purinergic-mediated Calcium Homeostasis and Dopamine R~lease in PC 12 Cells: Effect of Ethanol

  • Kim, Won-Ki
    • 한국생물물리학회:학술대회논문집
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    • 한국생물물리학회 1997년도 학술발표회
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    • pp.16-16
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    • 1997
  • Extracelluar ATP evokes many biological processes, including neuronal excitation and neurotransmitter secretion, through activation of purinergic P2 receptors. Although excitatory and inhibitory receptor-operated channels (ROC) and voltage-dependent calcium channels (VDCC) have been reported to be altered by acute and chronic exposure to ethanol, little is known of the ethanol effects on purinergic receptor-operated channels in neuronal cells.(omitted)

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한국적 의학 기준에 근거한 고혈압환자의 Angiotensin II Receptor Blockers와 Calcium Channel Blockers의 약물 평가 (Evaluation of Therapeutic Differences of Angiotensin II Receptor Blockers and Calcium Channel Blockers Among Hypertensive Patients Classified by Oriental Traditional Way)

  • 이옥상;천영주;예경남;윤희영;김정태;이윤정;임성실
    • 약학회지
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    • 제58권2호
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    • pp.141-149
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    • 2014
  • Background: Oriental lifestyle for treating diseases has been developed and well-accepted for a long time among Koreans. Sasang Constitution theory, originated from Korean traditional medicine, suggests that medication treatment should be differentiated by each patient's body classification (So-yang [SY], So-eum [SE], Tae-yang [TY], and Tae-eum [TE]), in contrary to western medicine's theory that medication should be applied equally by disease indication without such classification. However, the pharmacotherapeutic outcomes of these theories have not been compared to date. In this study, we aimed to compare the two theories by evaluating blood pressure (BP), which is lowered as a therapeutic outcome, among hypertensive patients taking angiotensin II receptor blockers (ARBs) or calcium channel blockers (CCBs), two most commonly used antihypertensive classes in Korea. Methods: From April 2006 to June 2012, we retrospectively collected data on hypertensive patients with Sasang Constitution classification at Kyunghee University Hospital at Gangdong, one of the East-West collaborative medical centers in Korea. We collected information on age, gender, underlying diseases, antihypertensive drugs (ARB, CCB, ARB+CCB), and BP by reviewing the electronic medical records. We excluded patients with missing blood pressure at baseline or follow-up, or those who had a change in their antihypertensive drug class during follow-up. Results: We selected a total of 573 patients (SY: 165, SE: 158, TY: 0, TE: 250). Baseline BPs were on average 139.0/82.0 mmHg for SY, 137.8/78.5 mmHg for SE, and 138.7/79.2 mmHg for TE. In all three groups, CCBs were the most prescribed, followed by combination therapy with ARB+CCB, then ARBs. BP reduction after 1 month of initial medication was significantly different among the drug classes, but not in Sasang constitutional classification (ARB [SY: -12.4/-4.7, SE: -12.3/-2.5, TE: -8.6/-1.8], CCB [SY: -12.3/-5.4, SE: -13.0/-2.3, TE: -10.8/-6.0], ARB+CCB [SY: -15.6/-6.7, SE: -18.4/-8.1, TE: -20.2/-6.7], drug [$P{\leq}0.05$/P>0.05], constitutional type [P>0.05/P>0.05]). Conclusion: We observed significant differences in reduction of blood pressure by classes of drugs (ARB+CCB>CCB>ARB) but not by Sasang constitutional classification. Therefore, current approach of antihypertensive pharmacotherapy assisted by Western medicine is appropriate for treatment of hypertension. However, further larger scale or prospective studies are required in order to confirm these results.

사람 혈청 트란스페린수용체의 단클론 항체를 이용한 방사면역측정과 철영양상태의 진단 (Immunoradiometric Assay using Monoclonal Antibody Against Human Serum Transferrin Receptor for Diagnosis of Iron Deficiency)

  • 김승렬
    • Journal of Nutrition and Health
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    • 제29권9호
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    • pp.971-980
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    • 1996
  • The soluble transferrin receptor(TfR) in human serum has been shown recently to be a truncated form of intact membrane bound receptor containing most of the extracellular domain. We purfied the transferin-free TfR from human serum by immounoaffinity chromatography which produced the single protein identity in high resolution gel chormatography. The monoclonal antibodies(MAb) against purifed serum TfR were produced by fusion of spleen cells o fimmunized Balb/c mice and SP2 cells. Ten hybrids producing MAb specific for serum TfR were identifed and determine their iostypes. A immunoraddiometric assay (IRMA) for serum TfR was established using two monoclonal IgG1 antibodies as the coating and indicator antibodies on the bosis of their suitability in sandwich IRMA of serum TfR. The mean serum TfR levels in the 15 normal male, 15 normal female, and 19 iron-deficient subjects were 5.4$\pm$0.98, 4.6$\pm$0/76, and 18.0$\pm$12.8mg/1, respectively, and the difference in mean values between normal and iron deficient subjects was significant(p=0.0005). There existed the inverse logarithmic relationship(r=-0.9336, p<0.0001) between the serum TfR and ferritin levels.

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Higher Ki67 Expression is Associates With Unfavorable Prognostic Factors and Shorter Survival in Breast Cancer

  • Kilickap, Saadettin;Kaya, Yalcin;Yucel, Birsen;Tuncer, Ersin;Babacan, Nalan Akgul;Elagoz, Sahande
    • Asian Pacific Journal of Cancer Prevention
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    • 제15권3호
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    • pp.1381-1385
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    • 2014
  • Background: The prognostic value of the Ki67 expression level is yet unclear in breast cancer. The aim of this study was to investigate the association between Ki67 expression levels and prognostic factors such as grade, Her2 and hormone receptor expression status in breast cancers. Materials and Methods: Clinical and pathological features of the patients with breast cancer were retreived from the hospital records. Results: In this study, 163 patients with breast cancer were analyzed, with a mean age of $53.4{\pm}12.2$ years. Median Ki67 positivity was 20% and Ki67-high tumors were significantly associated with high grade (p<0.001), lymphovascular invasion (p=0.001), estrogen receptor (ER) negativity (p=0.035), Her2 positivity (p=0.001), advanced stage (p<0.001) and lymph node positivity (p<0.003). Lower Ki67 levels were significantly associated with longer median relapse-free and overall survival compared to those of higher Ki67 levels. Conclusions: High Ki67 expression is associated with ER negativity, Her2 positivity, higher grade and axillary lymph node involvement in breast cancers. The level of Ki67 expression is a prognostic factor predicting relapse-free and overall survival in breast cancer patients.

Association of Human Epidermal Growth Factor Receptor-2 Expression and Clinicopathological Findings in Patients with Colorectal Cancer

  • Karaca, Halit;Deniz, Kemal;Berk, Veli;Inanc, Mevlude;Ozkan, Metin
    • Asian Pacific Journal of Cancer Prevention
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    • 제13권12호
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    • pp.6221-6225
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    • 2012
  • Background: To determine the frequency of HER-2 overexpression in colorectal cancer (CRC) patients, and to explore the relationship between clinicopathological prognostic factors and their effects on survival, based on immunohistochemistry (IHC) and fluorescent in situ hybridization (FISH) analysis. Materials and Methods: The study included 80 patients with a histologically proven diagnosis of CRC that received adjuvant FOLFOX-4 chemotherapy at our department between March 2006 and September 2010. Patient data were analyzed retrospectively. Results: The median follow-up period and age of the patients were 24 months and 59 years, respectively. In immunohistochemical staining, 3+ staining was found in 2 patients (2.5%) while 2+ was in 13 (16%). FISH for HER-2 was performed for all of these 15 patients; samples which were 3+ showed positivity but the ones with 2+ were negative. There was no significant correlation between HER-2 expression and age, gender, tumor localization, histological subtype, grade, lymphovascular and perineural invasion, or pTN stage (P>0.05), even when the patients with HER-2 overexpression were analyzed separately. There was also no significant relationship between progression-free survival (PFS) and overall survival (OS), and HER-2 expression, gender, tumor localization, obstruction-perforation, bleeding, histological type, grade, lymphovascular and perineural invasion, or pT staging (P>0.05); however, there was a significant relationship between lymph node involvement, and PFS and OS (P<0.05). Conclusions: Evaluation of HER-2 overexpression in a more comprehensive, multi-center, prospective trial with standardized methods will be an appropriate approach.

ACTIVATION OF P38 MAP KINASE AND AP-1 DURING THE PROMOTION OF NEURITE EXTENSION OF PC-12 CELLS BY 15-DEOXY-$\Delta$12,14-PROSTAGLANDIN J2

  • Song, Y.S.;Oh. J.H.;Park, K.S.;Jung, K.M.;Lee, M.K.;Jung, H.K.;Jung, S.Y.;Hong, J.T.
    • 한국독성학회:학술대회논문집
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    • 한국독성학회 2002년도 Current Trends in Toxicological Sciences
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    • pp.110-110
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    • 2002
  • 15-Deoxy-$\Delta$12, 14-prostaglandin J2 (15-deoxy-PGJ2), a naturally occurring ligand activates the peroxisome proliferator-activated receptor-$\gamma$(PPAR-$\gamma$). It was known to have promoting ability of nerve growth factor(NGF)-induced neurite extension. However, it is not clear yet as to what signaling pathway is involved in its promoting ability of neurite extension.(omitted)

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