• Title/Summary/Keyword: Activated factor X

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Purinergic regulation of calcium signaling and exocytosis in rat prostate neuroendocrine cells

  • Kim, Jun-Hee;Kim, Mean-Hwan;Koh, Duk-su;Park, So-Jung;Kim, Soo-Jung;Nam, Joo-Hyun;Lee, Jee-Eun;Uhm, Dae-Yong;Kim, Sung-Joon
    • Proceedings of the Korean Biophysical Society Conference
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    • 2003.06a
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    • pp.54-54
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    • 2003
  • Prostate gland contains neuroendocrine cells (PNECs) are playing important roles in physiological and pathophysiological processes of the prostate gland. Here, we investigated the role of purinoceptors in PNECs freshly isolated from rat ventral prostate (RPNECs) that show immunoreactivity to chromogranin A. Fura-2 ratiometry revealed that ATP evokes both fast Ca$\^$2+/ influx and store Ca$\^$2+/ release in RPNECs. A whole-cell patch clamp study demonstrated fast inactivating cationic current activated by ATP or by ${\alpha}$,${\beta}$-MeATP, which was blocked by ATP-TNP. The activation of P2X inward current was tightly associated with a sharp increase in [Ca$\^$2+/]$\sub$c/. The presence of P2X1/3 subtypes were proved by RT-PCR analysis. For the stored Ca$\^$2+/ release, ATP and UTP showed similar effects, suggesting the dominant role or P2Y2 subtypes, also confirmed by RT-PCR. Both P2X (${\alpha}$,${\beta}$-MeATP) and P2Y (UTP) stimulation induced changes in the cell morphology (initial shrinkage and blob formation on the surface) reversibly. Exocytotic membrane trafficking events were monitored with the membrane-bound fluorescent dye, FM1-43 using confocal microscopy. In spite of the similar Ca$\^$2+/ responses, UTP was far less effective in triggering exocytosis than ${\alpha}$,${\beta}$ -MeATP. Since serotonin is reportedly stored in the secretory granule of PNECs, we directly examined whether the aforementioned agonists elicit release of serotonin using carbon fiber electrode-amperometry. In accordance with the results of FM1 -43 experiments, ${\alpha}$,${\beta}$-MeATP efficiently evoke serotonin secretion while not with UTP. In summary, the P2X-mediated Ca$\^$2+/ influx plays crucial roles in the exocytosis of RPNECs. Although a global increase in [Ca$\^$2+]$\sub$c/ might be related with the morphological changes, a sharp rise of [Ca$\^$2+/]$\sub$c/ in the putative sub-plasmalemmal ‘microdomains’ might be a decisive factor for the exocytosis.

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Enhancement of Tumor Response by MEK Inhibitor in Murine HCa-I Tumors (C3H/HeJ 마우스 간암에서 MEK 억제제에 의한 방사선 감수성 향상 효과)

  • Kim, Sung-Hee;Seong, Jin-Sil
    • Radiation Oncology Journal
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    • v.21 no.3
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    • pp.207-215
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    • 2003
  • Purpose: Extracellular signal-regulated kinase (ERK), which is part of the mitogen-activated protin kinase cascade, opposes initiation of the apoptotic cell death which is programmed by diverse cytotoxic stimuli. In this regard, the inhibition of ERK may be useful in improving the therapeutic efficacy of established anticancer agents. Materials and Methods: Murine hepatocarcinoma, HCa-I is known to be highly radioresistant with a TCD50 (radiation dose yield in $50\%$ cure) of more than 80 Gy. Various anticancer drugs have been found to enhance the radioresponse of this particular tumor but none were successful. The objective of this study was to explore whether the selective inhibition of MEK could potentiate the antitumor efficacy of radiation in vivo, particularly in the case on radioresistant tumor. C3H/HeJ mice hearing $7.5\~8\;mm$ HCa-I, were treated with PD98059(intratumoral injection of $0.16\;\mug/50\;\mul$). Results: Downregulation on ERK by PD98059 was most prominent 1h after the treatment. In the tumor growth delay assay, the drug was found to Increase the effect of the tumor radioresponse with an enhancement factor (EF) of 1.6 and 1.87. Combined treatment of 25 Gy radiation with PD98059 significantly increased radiation induced apoptosis. The peak apoptotic index (number on apoptotic nuclei in 1000 nuclei X100) was $1.2\%$ in the case of radiation treatment alone, $0.9\%$ in the case of drug treatment alone and $4.9\%,\;5.3\%$ in the combination treatment group. An analysis of apoptosis regulating molecules with Western blotting showed upregulation of p53, p$p21^{WAF1/CIP1}\;and\;Bcl-X_s$ in the combination treatment group as compared to their levels in either the radiation alone or drug alone treatment groups. The level of other molecules such as $Bcl-X_L4, Bax and Bcl-2 were changed to a lesser extent. Conclusion: The selective inhibition of MEK in combination with radiation therapy may have potential benefit in cancer treatment.

MILD HEMOPHILIA A DETECTED BY DELAYED BLEEDING: A CASE REPORT (지연된 출혈로 밝혀진 경증 혈우병 A에 대한 증례 보고)

  • Song, Je-Seon;Choi, Byung-Jai;Kim, Seong-Oh;Lee, Jae-Ho;Son, Heung-Gyu;Kim, Hyung-Jun;Choi, Hyung-Jun
    • The Journal of Korea Assosiation for Disability and Oral Health
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    • v.4 no.2
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    • pp.73-76
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    • 2008
  • Hemophilia A is an X-linked disorder resulting from a deficiency in factor VIII. Hemophilia A is classified into severe, moderate, and mild forms. A 12-month old boy visited for uncontrolled bleeding on the upper labial frenum area. Spontaneous bleeding occurred about 13 days ago. We sutured the wound. However, the patient returned after 6 days with large hematoma formation. Consultation to the hematologist and laboratory examination were carried out. Activated PTT was elongated and factor VIII was only 6%. He was diagnosed as mild hemophilia A. Fresh frozen plasma (FFP) was provided and hemostasis was achieved. At 5-day check-up, no more bleeding was observed. For the management of prolonged bleeding in hemophilia A patient, not only local hemostatic measures, but factor VIII replacement therapy, antifibrinolytics, and Desmopressin are also available. Mild hemophilia A often occurs without either familial or medical history. It is often first detected by dental trauma.

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Research on Anti-lipogenic Effect and Underlying Mechanism of Laminaria japonica on Experimental Cellular Model of Non-alcoholic Fatty Liver Disease (비알코올성 지방간 세포 모델에서 곤포의 효능과 기전 연구)

  • Kim, So-Yeon;Kwon, Jung-Nam;Lee, In;Hong, Jin-Woo;Choi, Jun-Yong;Park, Seong-Ha;Kwun, Min-Jung;Joo, Myung-Soo;Han, Chang-Woo
    • The Journal of Internal Korean Medicine
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    • v.35 no.2
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    • pp.175-183
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    • 2014
  • Objectives : We tried to uncover the anti-lipogenic effect and underlying mechanism of Laminaria japonica on an experimental cellular model of non-alcoholic fatty liver disease. Methods : Ethanol extract of Laminaria japonica (LJ) was prepared. Intracellular lipid content of palmitate-treated HepG2 cells was evaluated with or without LJ treatment. We measured the effects of LJ on liver X receptor ${\alpha}$ ($LXR{\alpha}$) and sterol regulatory element-binding transcription factor-1c (SREBP-1c) expression, transcription level of lipogenic genes, including acetyl-CoA carboxylase (ACC), fatty acid synthase (FAS), stearoyl-CoA desaturase-1 (SCD-1), and nuclear factor erythroid 2-related factor 2 (Nrf2) activation in HepG2 cells. Results : LJ markedly attenuated palmitate-induced intracellular lipid accumulation in HepG2 cells. LJ suppressed $LXR{\alpha}$-dependent SREBP-1c activation, and SREBP-1c mediated induction of ACC, FAS, and SCD-1. Furthermore, LJ activated Nrf2, which plays an important cytoprotective role in non-alcoholic fatty liver disease. Conclusions : Our study suggests that LJ has the potential to alleviate hepatic lipid accumulation, and this effect was mediated by inhibiting the $LXR{\alpha}$-SREBP-1c pathway that leads to hepatic steatosis. In addition, the anti-lipogenic potential may, at least in part, be associated with activation of Nrf2.

Effect of Snake Venom Toxin on Inhibition of Colorectal Cancer HT29 Cells Growth via Death Receptors Mediated Apoptosis

  • Shim, Yoon Seop;Song, Ho Sueb
    • Journal of Acupuncture Research
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    • v.31 no.2
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    • pp.87-98
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    • 2014
  • Objectives : We investigated whether snake venom toxin(SVT) from Vipera lebetina turanica sensitizes HT29 human epithelial colorectal cancer cells to tumor necrosis factor(TNF)-related apoptosis-inducing ligand(TRAIL) induced apoptosis in cancer cells. Methods : Cell viability assay was used to assess the inhibitory effect of TRAIL on cell growth of HT29 human colorectal cancer cells. And 6-diamidino-2-phenylindole(DAPI), terminal deoxynucleotidyl transferase mediated dUTP nick end labeling assay(TUNEL) staining assay were used to evaluate cell-apoptosis. Western blot analysis were conducted to observe apoptosis related proteins and death receptor. To assess whether the synergized inhibitory effect of SVT and TRAIL on reactive oxygen species(ROS) generation was reversed by strong anti-oxidative agent. Results : SVT with TRAIL inhibited HT29 cell growth different from TRAIL alone. Consistent with cell growth inhibition, the expression of TRAIL receptors; Expression of death receptor(DR)4 and DR5 was significantly increased and intrinsic pro-apoptotic cleaved caspase-3, -9 was subsequently increased together with increase of Bax/Bcl-2 ratio and extrinsic pro-apototic caspase-8 was also activated. In addition, the expression of anti-apoptotic survival proteins, a marker of TRAIL resistance(eg, cFLIP, survivin, X-linked inhibitor of apoptosis protein(XIAP) and Bcl-2) was suppressed by the combination treatment of SVT and TRAIL. Pretreatment with the ROS scavenger N-acetylcysteine abolished the SVT and TRAIL-induced upregulation of DR4 and DR5 expression and expression of the intrinsic pro-apoptotic caspase-3 and-9. Conclusion : The collective results suggest that SVT facilitates TRAIL-induced apoptosis in $HT_{29}$ human epithelial colorectal cancer cells through up-regulation of the TRAIL receptors; DR4 and DR5 and consecutive induction of bilateral apoptosis via regulating apoptosis related proteins.

A Study on the Health Instruction of School Nurses and Its Related Factors (양호교사(養護敎師)의 보건수업(保健授業) 실시정도(實施程度)와 이의 관련요인(關聯要因) 분석(分析) - 서울시 초(初)·중등(中等) 양호교사(養護敎師)를 중심(中心)으로 -)

  • Lee, Weon You
    • Journal of the Korean Society of School Health
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    • v.3 no.2
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    • pp.63-87
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    • 1990
  • The purpose of this study was to investigate the performance rate of regular health instruction by school nurses and its related factors. These data were collected by questionnaire in Mar. 1990. Total 986 school nurses who have been working primary, secondary schools in Seoul were given the questionnaire, and 672 school nurses' respectively, were finally analyzed. Analysis of the data was done utilizing SPSS/PC+for percentage, frequency, mean, $X^2$-test, t-test, ANOVA, correlation, regression. The major findings of this study were as follows : 1. Thc performance rate of regular health instruction by school nurses was 77.7% of 552 respondents. 2. The school nurses in primary school(87.0%) were more likely to perform regular health instruction than the school nurses in secondary school (63.8%). 3. The teachers of regular health instruction were more likely to display favorable attitudes toward need of health instruction. The school level was not statistically significant(P>0.05). 4. The evidence from the data analysis supported the proposition that the teachers of regular health instruction had more positive attitude toward the establishment of health subject(80.3%). The school level was statistically significant(P<0.05). 5. The school principal was a highly significant factor as it affected the performance rate of regular health instruction by school nurses. The school level was statistically significant(P<0.001). 6. The length of past career, the degree of self confidence on health instruction, the need of health instruction did affect the performance rate of regular health instruction by school nurses. On the basis of the above mentioned, so as for regular health instruction of school to be activated more throughly, regular class must be introduced in the form of establishment of health subject, enhancement of quality on the part of school nurses, made and various methods for enhancing perception toward the regular health instruction on the part of school principal are to be carried out.

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Inhibitory Effects of Bojungchiseub-tang on Adipocyte Differentiation and Adipogenesis in 3T3-L1 Preadipocytes (보중치습탕이 3T3-L1 지방전구세포의 분화 및 지방생성 억제에 미치는 영향)

  • Lee, Soo Jung;Kim, Won Il;Kang, Kyung Hwa
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.28 no.3
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    • pp.288-295
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    • 2014
  • Bojungchiseub-tang (BJCST) has been used in symptoms and signs of edema, dampness-phlegm, kidney failure, and so on. BJCST is also expected to have strong anti-obesity activities. However, little is known about the mechanisms of its inhibitory effects on adipocyte differentiation and adipogenesis. In the present study, we examined the effects and mechanism of BJCST on transcription factors and adipogenic genes of 3T3-L1 preadipocytes to understand its inhibitory effects on adipocyte differentiation and adipogenesis. Our results showed that BJCST significantly inhibited differentiation and adipogenesis of 3T3-L1 preadipocytes in a dose-dependent manner. To elucidate the mechanism of the effects of BJCST on lowering lipid content in 3T3-L1 adipocytes, we examined whether BJCST modulate the expressions of transcription factors to induce adipogenesis and adipogenic genes related to regulate accumulation of lipids. As a result, the expression of steroid regulatory element-binding protein (SREBP)1, cytidine-cytidine-adenosine-adenosine-thymidine (CCAAT)/enhancer binding proteins ${\alpha}$ ($C/EBP{\alpha}$), $C/EBP{\beta}$, $C/EBP{\delta}$, and peroxisome proliferator-activated receptor ${\gamma}$ ($PPAR{\gamma}$) genes, which induce the adipose differentiation, liver X receptor $(LXR){\alpha}$ and fatty acid synthase (FAS) genes, which induce lipogenesis and adipose-specific aP2, Adipsin, lipoprotein lipase (LPL), CD36, TGF-${\beta}$, leptin and adiponectin genes, which compose fat formation were decreased. BJCST also reduced the expression of acyl CoA oxidase (ACO) and uncoupling protein (UCP) genes related to lipid oxidation. In conclusion, BJCST could regulate transcript factor related to induction of adipose differentiation and inhibited the accumulation of lipids and expression of adipogenic genes.

Functional Implications of Transporters Under Nitrosative Stress Conditions

  • Yu, Kyung-Ha;Maeng, Han-Joo;Chung, Suk-Jae
    • Journal of Pharmaceutical Investigation
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    • v.40 no.3
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    • pp.139-153
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    • 2010
  • Nitrosative stress is defined as pathophysiological conditions that are related to covalent modifications of proteins by nitration/nitrosylation by forms of nitrogen oxide ($NO_x$), leading to DNA damage, ultimately, cell death. This type of stress condition appears to be associated with a number of disease states, including diabetes, inflammation and neurodegenerative diseases. Since these pathological conditions are frequently chronic in nature and, thus, require long-term treatment, changes in pharmacokinetics are likely to affect the therapy. Transporters are membrane proteins that facilitate the movement of substrates, including drugs, across plasma membranes of epithelial / endothelial cells. Since it is now increasingly evident that transporters are pharmacokinetically significant, functional alteration of transporters by this stress condition may have therapeutic relevance. In this review, experimental techniques that are used to study both in vivo and in vitro nitrosative stress are summarized and discussed, along with available literature information on the functional implication of transporters under conditions of nitrosative stress conditions. In the literature, both functional induction and impa irment were apparently present for both drug transporter families [i.e., ATP-binding cassette (ABC) and solute carrier families (SLC)]. Furthermore, a change in the function of a certain transporter appears to have temporal dependency by impairment in the early phase of nitrosative stress and induction thereafter, suggesting that the role of nitrosative stress is complex in terms of functional implications of the transporters. Although the underlying mechanisms for these alterations are not fully understood, protein nitration/nitrosylation appears to be involved in the functional impairment whereas transcript factor(s) activated by nitrosative stress may play a role, at least in part, in functional induction. Interestingly, functional induction under conditions of nitrosative stress has not been observed for SLC transporters while such impairment has been documented for both ABC and SLC transporters. Further investigations appear to be necessary to fully delineate the underlying reasons for these differences on the impact and importance of nitrosative stress conditions.

Hydrothermal Synthesis of Smectite from Dickite (딕카이트로부터 스멕라이트의 수열합성)

  • Ryu Gyoung-Won;Jang Young-Nam;Bae In-Kook;Chae Soo-Chun;Choi Sang-Hoon
    • Journal of the Mineralogical Society of Korea
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    • v.17 no.3
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    • pp.267-275
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    • 2004
  • A hydrothermal process was used to synthesize dioctahedral smectite from dickite [$A1_2$$Si_2$$O_{5}$ $(OH)_4$], Dickite was previously activated by heating at $800^{\circ}C$ far 4 hours with $Na_2$$CO_3$. After the heat-treatment, $SiO_2$ was added for stoichiometry, The autoclaving was carried out in closed stainless steel vessel (about 1 liter) at the condition of various temperature, pressure, time etc. High quality smectite could be obtaind by heating at $290^{\circ}C$ under the pressure of 60 kgf/$\textrm{cm}^2$ for 48 hours. This experiment reveals that pH of the solution was an important factor and should be maintained at 10 to 11 for the formation of dioctahedral smectite. The synthesized smectite was identified as Na-beidellite by the treatment of ethylene glycol and Greene-Kelly test.

Polymorphisms in the Promoter Region of the Chinese Bovine PPARGC1A Gene

  • Li, M.J.;Liu, M.;Liu, D.;Lan, X.Y.;Lei, C.Z.;Yang, D.Y.;Chen, H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.26 no.4
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    • pp.483-487
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    • 2013
  • The peroxisome proliferator-activated receptor gamma coactivator-1 alpha protein, encoded by the PPARGC1A gene, plays an important role in energy homeostasis. The genetic variations within the PPARGC1A gene promoter region were scanned in 808 Chinese native bovines belonging to three cattle breeds and yaks. A total of 6 SNPs and one 4 bp insertion variation in the promoter region of the bovine PPARGC1A gene were identified: SNP -259 T>A, -301_-298insCTTT, -915 A>G, -1175 T>G, -1590 C>T, -1665 C>T and -1690 G>A, which are in the binding sites of some important transcription factors: sex-determining region Y (SRY), myeloid-specific zinc finger-1 (MZF-1) and octamer factor 1(Oct-1). It is expected that these polymorphisms may regulate PPARGC1A gene transcription and might have consequences at a regulatory level.