• Title/Summary/Keyword: P release and uptake

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Functional Characterization and Regional Expression of Dopamine Transporter (도파민 수송체의 기능적 특성 및 발현에 관한 연구)

  • 이상훈;이송득;성기욱;이동섭;이용성;고재경
    • YAKHAK HOEJI
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    • v.39 no.2
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    • pp.161-168
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    • 1995
  • Brain dopamine systems play a central role in the control of movement, hormone release, and many complex behavior. The action of dopamine at its synapse is terminated predominately by high affinity reuptake into presynaptic terminals by dopamine transporter (DAT). The dopamine transporter(DAT) is membrane protein localized to dopamine-containing nerve terminals and closely related with cocaine abuse, Parkinsonism, and schizophrenia. In present study, the recombinant plasmid pRc/CMV-DAT, constructed by subcloning of a cDNA encoding a bovine DAT into eukaryotic expression vector pRc/CMV, was stably transfected into CV-1 cells(monkey kidney cell line). The DAT activities in the cell lines selected by Geneticin$^{R}$ were determined by measuring the uptake of $[^3H]$-dopamine. The transfected cell lines showed 30-50 fold higher activities than untransfected CV-1 cell line, and this result implies that DAT is well expressed and localized in transfected cells. The transfected cells accumulated $[^3H]$-dopamine in a dose-dependent manner with a $K_{m}$ of 991.6nM. Even though high doses of norepinephrine, epinephrine, serotonin, and choline neurotransmitters inhibited the uptake of $[^3H]$-dopamine, DAT in transfected cell line was proven to be much more specific to dopamine. The psychotropic drugs such as GBR12909, CFT, normifensine, clomipramine, desipramine, and imipramine inhibited significantly the dopamine uptake in tissue culture cells stably transfected with DAT cDNA. Radioactive in situ hybridization was done to map the cellular localization of DAT mRNA-containing cells in the adult rat central nervous system. The strong hybridization signals were detected only in the substantia nigra pars compacta and ventral tegmental area. The restricted anatomical localization of DAT mRNA-containing cells confirms the DAT as a presynaptic marker of dopamine-containing cells in the rat brain.

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Effect of Piperine on Peripheral Sympathetic Nervous System in Isolated Vas deferens of Rat (Piperine이 적출 백서 정관내의 교감신경계에 미치는 영향)

  • Eun, Jae-Soon
    • YAKHAK HOEJI
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    • v.32 no.1
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    • pp.55-61
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    • 1988
  • To elucidate one of the effect of piperine on the peripheral sympathetic nervous system, influence of piperine upon the contractile action of norepinephrine, methoxamine and tyramine as well as uptake and release of $[^{3}H]-norepinephrine$ has been investigated in naive and chronic piperine-treated vas deferens of rats. $pA_2$ value for ${\alpha}_1-adrenoceptor$ of phentolamine was significantly increased. Chronic piperine-treated group was markedly shown increased efflux of $[^{3}H]-norepinephrine$ and muscular tension, but was not affected the neuronal up-take and release of $[^{3}H]-norepinephrine$. It can be concluded that potentiation of the effect of norepinephrine by acute and chronic piperine treated group may be due to the change of affinity of ${\alpha}_1-adrenoceptor$, and partly due to possible modification of storage mechanism.

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Immunomodulatory Effect of Pueraria lobata on the Functional Activation of Macrophages by Lipopolysaccharide Treatment (그람음성균 유래 lipopolysaccharide로 활성화된 대식세포의 기능 수행시 갈화 [Pueraria lobata (Willd.) Ohwi]의 조절작용)

  • Shen, Ting;Kim, Jong-Dai;Cho, Jae-Youl
    • Korean Journal of Medicinal Crop Science
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    • v.17 no.1
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    • pp.8-14
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    • 2009
  • Pueraria lobata (Willd.) Ohwi was investigated to check its modulatory effects on the activation of macrophages upon inflammatory conditions treatment. For this purpose, we examined several inflammatory responses such as nitric oxide (NO) production, reactive oxygen species (ROS) generation, cytoprotection and phagocytosis under the treatment of methanol extract from P. lobata (Pl-ME). Pl-ME dose-dependently blocked NO production in lipopolysaccharide (LPS)- stimulated RAW264.7 cells but not sodium prusside (SNP)-generated NO release. The NO inhibition seemed to be due to blocking inducible NO synthase (iNOS), since Pl-ME suppressed its expression in a NF-${\kappa}B$-independent manner. Similarly, this extract also effectively protected RAW264.7 cells from LPS-induced cytotoxicity. However, Pl-ME did not block ROS generation and rather it enhanced. Finally, this extract negatively modulated FITC-dextran uptake. Therefore, our data suggested that Pl-ME may be involved in negatively regulating some macrophage-mediated inflammatory responses such as NO production and phagocytic uptake.

COMPARATIVE STUDY OF FLUORIDE RELEASE AND RE-UPTAKE OF SEVERAL RESTORATIVE MATERIALS (수종 수복재에 불소 도포제 적용 후 불소유리에 관한 비교연구)

  • Kim, Soo-Yeon;Kim, Jong-Soo;Yoo, Seung-Hoon
    • Journal of the korean academy of Pediatric Dentistry
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    • v.34 no.3
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    • pp.408-419
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    • 2007
  • In the child, it is very important that he/she will have the ability to suppress aesthetic restorative materials of secondary caries. With the representative preventive material against caries, the importance of fluoride is more emphasized. This study examined the differences in fluoride release and re-uptake among some restorative materials, following a treatment of APF gel and fluoride varnish. The surface roughness was observed under scanning electron microscope. Studying this will provide for the research to find effective restorative materials and fluoride type in tooth caries prevention. It is applied from presence at a clinic that restorative materials are resin, flowable resin, compomer and glass ionomer. Fluoride release was measured at 24-hour intervals for 7 days, 3-day intervals from 8th to 38th day using an ion-selective electrode and analyzer. Then, the materials were treated with the fluoride gel and fluoride varnish respectively, fluoride release was measured and specimens were evaluated under scanning electron microscope for 4 weeks. It was concluded that 1. Fluoride was released for 38 days from restorative materials under 1 ppm in case of flowable resin, 1-2 ppm in compomer and 2-8 ppm in glass ionomer, a few of fluoride was released after 45 days 2. Fluoride has more releasing after application of APF gel than fluoride varnish. Fluoride re-uptake was observed under 0.6-0.2 ppm in fluoride varnish and 0.6-2.6 ppm in APF gel after starting the procedure one day(p<0.05). For the remaining 4 weeks, they demonstrated a similar release. 3. Specimens were evaluated under scanning electron microscope. Applied fluoride in the experimental group surface was rougher than the control group that did not receive fluoride application. Fluoride varnish group had a smoother surface than both the APF gel group and the varnish APF gel group that received a fluoride application.

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Effect of Podophyllotoxin Conjugated Stearic Acid Grafted Chitosan Oligosaccharide Micelle on Human Glioma Cells

  • Wang, Geng Huan;Shen, He Ping;Huang, Xuan;Jiang, Xiao Hong;Jin, Cheng Sheng;Chu, Zheng Min
    • Journal of Korean Neurosurgical Society
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    • v.63 no.6
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    • pp.698-706
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    • 2020
  • Objective : To study the physiochemical characteristics of podophyllotoxin (PPT) conjugated stearic acid grafted chitosan oligosaccharide micelle (PPT-CSO-SA), and evaluate the ability of the potential antineoplastic effects against glioma cells. Methods : PPT-CSO-SA was prepared by a dialysis method. The quality of PPT-CSO-SA including micellar size, zeta potential, drug encapsulation efficiency and drug release profiles was evaluated. Glioma cells were cultured and treated with PPT and PPT-CSO-SA. The ability of glioma cells to uptake PPT-CSO-SA was observed. The proliferation of glioma cells was determined by 3-[4, 5-dimethyl-2-thiazolyl]-2, 5-diphenyl-2H-tetrazolium bromide (MTT) assay. The apoptosis and morphology of U251 cells were observed by 4',6-Diamidino-2-phenylindole dihydrochloride (DAPI) dye staining. Cell cycle analysis was performed by flow cytometry. The migration ability of U251 cells was determined by wound healing test. Results : PPT-CSO-SA had nano-level particle size and sustained release property. The encapsulation efficiency of drug reached a high level. The cellular uptake percentage of PPT in glioma cells was lower than that of PPT-CSO-SA (p<0.05). The inhibitory effect of PPT-CSO-SA on glioma cells proliferation was significantly stronger than that of PPT (p<0.05). The morphologic change of apoptosis cell such as shrinkage, karyorrhexis and karyopyknosis were observed. The percentage of U251 cells in G2/M phase increased significantly in the PPT-CSO-SA group compared with PPT group (p<0.05). Compared with the PPT group, the cell migration ability of the PPT-CSO-SA group was significantly inhibited after 12 and 24 hours (p<0.05). Conclusion : PPT-CSO-SA can effectively enhance the glioma cellular uptake of drugs, inhibit glioma cells proliferation and migration, induce G2/M phase arrest of them, and promote their apoptosis. It may be a promising anti-glioma nano-drug.

Isolation and Characterization of Phosphorus Accumulating Acinetobacter CW3 (인 축적균 Acinetobacter CW3의 분리 및 특성)

  • 심성훈;류원률;이영호;김정목;조무환
    • KSBB Journal
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    • v.14 no.1
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    • pp.71-75
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    • 1999
  • A highly effective phosphorus accumulating bacterium named Acinetobacter CW3 was isolated from the nature by using Winogradsky columns. The optimal cultivation conditions of Acinetobacter CW3 in shaking flask were determined as $20^{\circ}C$, pH 7, 200rpm, 18.5mg $PO_4$-P/L. Acientobacter CW3 could remove phosphorus completely in 12hours for a batch culture at optimal cultivation condition. This bacterium could uptake phosphorus on aerobic condition and release it on anaerobic condition.

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Effects of mixing/aeration ratio and SRT on nutrient removal in SBR process (연속회분식반응조 공정에서 교반/폭기비와 SRT가 영양염류제거에 미치는 영향)

  • Jeon, Seok-Jun;Kim, Han-Soo
    • Journal of Korean Society on Water Environment
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    • v.18 no.3
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    • pp.291-301
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    • 2002
  • In this study, nutrients treatment by sequencing batch reactors(SBR) was performed. Nitrogen and phosphorus removal efficiencies were evaluated by changing SRT and mixing/aeration ratio. Not only nitrogen but also phosphorus removal patterns were investigated through track studies on 1 cycle. As SRT was fixed and mixing/aeration ratio was changed, maximum nitrogen removal efficiency was 87.6% at mixing/aeration ratio 0.67. Phosphorus removal efficiencies were more than 85.5% except no mixing condition. As mixing/aeration ratio was fixed and SRT was changed, nitrogen removal efficiencies were 70.5~79.8%, which represented slight changes, while phosphorus removal efficiencies were 49.0~97.3%, which represented sharply decreasing tendency at less than 20 day. Both phosphorus release rate k and maximum phosphorus release rate $P_{max}/M$ were are decreased as SRT was decreased, but they were not affected by mixing/aeration ratio. It was found that there is a linear relationship between ortho-phosphate uptake and maximum ortho-phosphate release.

Inositol 1,4,5-Trisphosphate-induced Increase in $Ca^{2+}-ATPase$ Activity in the Microsomes of Tracheal Epithelial Cells

  • Cho, Hyoung-Jin;Park, Sung-Shin;Kim, Young-Kee
    • The Korean Journal of Physiology
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    • v.29 no.2
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    • pp.269-277
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    • 1995
  • Membrane vesicles were prepared by differential centrifugation from epithelial cells of porcine trachea. Total activity of microsomal ATPases was measured spectrophotometrically by a coupled enzyme assay. The steady-state activity of the enzyme was $329{\pm}10$ nmol/min mg protein. Thapsigargin, a specific antagonist of intracellular $Ca^{2+}-ATPase$, inhibited about 50% of the activity, leaving $178{\pm}18\;nmol/min .mg$ protein (n=6), indicating that the $Ca^{2+}-ATPase$ is one of the major microsomal ATPases. The microsomes used in this study appeared to be tight-sealed vesicles since they showed saturation in $^{45}Ca^{2+}$ uptake experiments. Inositol 1,4,5-trisphosphate $InsP_{3}, 4\;{\mu}M$, an agonist of $InsP_{3}$-sensitive $Ca^{2+}$ release channel ($InsP_{3}$, receptor), and Ca-ionophore A23187 $(10\;{\mu}M)$ induced $^{45}Ca^{2+}$ releases of 20% and 50% of stored $^{45}Ca^{2+}$, respectively. The addition of $(10\;{\mu}M\;InsP_{3}$ also increased the microsomal ATPase activity from $282{\pm}8$ nmol/min mg protein to $334{\pm}21$ nmol/min . mg protein in the intact vesicles. Similar increase in the activity was observed by making microsomes leaky (uncoupling) using the Ca-ionophore A23187. ;$InsP_{3}-induced$ effects were blocked by either thapsigargin or heparin suggesting that: 1) the $InsP_{3}-induced$ increase in ATPase activity is mediated by microsomal $Ca^{2+}-ATPase$, and 2) dissipation of $Ca^{2+}$ gradient across the microsomal membrane is responsible for the $InsP_{3}-induced$ effect. In order to test the dependence of the $Ca^{2+}-ATPase$ activity on the activity of $InsP_{3}-induced$ the activity of ATPases was monitored in various concentrations of free $Ca^{2+}$ using $EGTA-Ca^{2+}$ buffers. The $Ca^{2+}$-dependent biphasic change is the well-known character of $InsP_{3} receptor but not of microsomal $Ca^{2+}-ATPase$ in non-excitable cells; however, the activity of microsomal ATPase appeared biphasic and a maxim진 activity of $397{\pm}36nmol/min\;.mg$ protein was obtained in the solution containing 100 nM free $Ca^{2+}$. Below or above this concentration, the activity of ATPases was lower. These results strongly support a positive correlation of microsomal $Ca^{2+}-ATPase$ to the $InsP_{3}$ receptors in epithelial microsomes.

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Nutrients removal on Oxic/Anoxic time ratio in 2-stage-intermittent-aeration reactor (2단 간헐 포기조의 포기/비포기 시간비에 따른 영양염류 제거특성)

  • Kim, Hong Tae;Sin, Seok U;O, Sang Hwa;Gwon, Seong Hyeon
    • Journal of Environmental Science International
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    • v.13 no.7
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    • pp.675-680
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    • 2004
  • This study was conducted to remove organics and nutrients using 2 stage intermittent aeration reactor. First reactor, using suspended microbial growth in intermittent aeration instead of anaerobic reactor in the typical BNR process, used minimum carbon source to release P, and it was possible to reduce ammonia loading going to second reactor. In the second reactor, using moving media intermittent aeration, it was effective to reduce nitrate in non-aeration time by attached microorganisms having long retention time. In aeration time, nitrification and P uptake were taken place simultaneously. From the experiment, two major results were as follows. First, the removal of organics was more than 90%, and optimum aeration/non-aeration time ratio for organic removal was corresponded with aeration/non-aeration time ratio for nitrogen removal. Second, in the first reactor, optimum aeration/non-aeration time ratio was 15/75 (min.) because it was necessary to maintain 75 min. of non-aeration time to suppress of impediment of return nitrate and to lead release of phosphate. In the second reactor, optimum aeration/non-aeration time ratio was 45/90 (min.).

A Study on the Removal of Nitrogen and Phosphorus of Municipal Wastewater with Biological Coated Media (미생물 코팅 담체를 이용한 하수의 질소$\cdot$인 제거에 관한 연구)

  • Kim Young-Gyu;Cho Il-Hyoung
    • Journal of Environmental Health Sciences
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    • v.32 no.1 s.88
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    • pp.27-35
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    • 2006
  • This study was carried out to investigate the removal of nitrogen and phosphorus in municipal sewage depending on existence of biological coated media in BCM reactor. The reactor with biological coated media is the process combining $A_2/O$ process. The removal efficiencies for $COD_{Mn},\; BOD_5,\;SS$, T-N and T-P were $78\%,\;90.5\%,\;92.3\%,\;61.9\%,\;60.2\%$, respectively. The specific nitrification rate$(mgNO_3-N/gMLSS{\cdot}d)$ of Contact aeration basin was 52.2 and the specific denitrification rate$(mgNO_3N/gMLSS{\cdot}d)$ in anoxic basin was 95.1. Also, phosphorus release$(mgPO_4-P/gMLSS{\cdot}d)$ in Anaerobic basin was 71.8 and Phosphorus uptake$(mgPO_4-P/gMLSS{\cdot}d)$ in contact aeration was 27.1.