• Title/Summary/Keyword: $Ca^{2+}$ signal

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Cloning, Expression, and Characterization of a Highly Active Alkaline Pectate Lyase from Alkaliphilic Bacillus sp. N16-5

  • Li, Gang;Rao, Lang;Xue, Yanfen;Zhou, Cheng;Zhang, Yun;Ma, Yanhe
    • Journal of Microbiology and Biotechnology
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    • v.20 no.4
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    • pp.670-677
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    • 2010
  • An alkaline pectate lyase, Bsp165PelA, was purified to homogeneity from the culture broth of alkaliphilic Bacillus sp. N16-5. The enzyme showed a specific activity as high as 1,000 U/mg and had optimum activity at pH 11.5 and $50^{\circ}C$. It was composed of a single polypeptide chain with a molecular mass of 42 kDa deduced from SDS-PAGE, and its isoelectric point was around pH 6.0. It could efficiently depolymerize polygalacturonate and pectin. Characterization of product formation revealed unsaturated digalacturonate and trigalacturonate as the main products. The pectate lyase gene (pelA) contained an open reading frame (ORF) of 1,089 bp, encoding a 36-amino acids signal peptide and a mature protein of 326 amino acids with a calculated molecular mass of 35.943 Da. The deduced amino acid sequence from the pelA ORF exhibited significant homology to those of known pectate lyases in polysaccharide lyase family 1. Some conserved active-site amino acids were found in the deduced amino acid sequence of Bsp165PelA. $Ca^{2+}$ was not required for activity on pectic substrates.

Phorbol 12-Myristate 13-Acetate Enhances Long-Term Potentiation in the Hippocampus through Activation of Protein Kinase $C{\delta}$ and ${\varepsilon}$

  • Kim, Eung Chang;Lee, Myeong Jong;Shin, Sang Yep;Seol, Geun Hee;Han, Seung Ho;Yee, Jaeyong;Kim, Chan;Min, Sun Seek
    • The Korean Journal of Physiology and Pharmacology
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    • v.17 no.1
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    • pp.51-56
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    • 2013
  • Many intracellular proteins and signaling cascades contribute to the sensitivity of N-methyl-D-aspartate receptors (NMDARs). One such putative contributor is the serine/threonine kinase, protein kinase C (PKC). Activation of PKC by phorbol 12-myristate 13-acetate (PMA) causes activation of extracellular signal-regulated kinase (ERK) and promotes the formation of new spines in cultured hippocampal neurons. The purpose of this study was to examine which PKC isoforms are responsible for the PMA-induced augmentation of long-term potentiation (LTP) in the CA1 stratum radiatum of the hippocampus in vitro and verify that this facilitation requires NMDAR activation. We found that PMA enhanced the induction of LTP by a single episode of theta-burst stimulation in a concentration-dependent manner without affecting to magnitude of baseline field excitatory postsynaptic potentials. Facilitation of LTP by PMA (200 nM) was blocked by the nonspecific PKC inhibitor, Ro 31-8220 ($10{\mu}M$); the selective $PKC{\delta}$ inhibitor, rottlerin ($1{\mu}M$); and the $PKC{\varepsilon}$ inhibitor, TAT-${\varepsilon}V1$-2 peptide (500 nM). Moreover, the NMDAR blocker DL-APV ($50{\mu}M$) prevented enhancement of LTP by PMA. Our results suggest that PMA contributes to synaptic plasticity in the nervous system via activation of $PKC{\delta}$ and/or $PKC{\varepsilon}$, and confirm that NMDAR activity is required for this effect.

Quantitative Assay of Recombinant Hepatitis B Surface Antigen by Using Surface Plasmon Resonance Biosensor (Surface plasmon resonance 바이오센서를 이용한 재조합 B형 간염 표면항원의 정량분석)

  • Lee, E. K.;Ahn, S. J.;Yoo, C. H.;Ryu, K.;Jeon, J. Y.;Lee, H. I.;Choi, S. C.;Lee, Y. S.
    • KSBB Journal
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    • v.17 no.1
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    • pp.20-25
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    • 2002
  • We performed a basic experiment for rapid, on-line, real-time measurement of HBsAg by using a surface plasmon resonance biosensor to quantify the recognition and interaction of biomolecules. We immobilized the anti-HBsAg polyclonal antibody to the dextran layer on a CM5 chip surface which was pre-activated by N-hydroxysuccinimide for amine coupling. The binding of the HBsAg to the immobilized antibody was measured by the mass increase detected by the change in the SPR signal. The binding characteristics between HBsAg and its antibody followed typical monolayer adsorption isotherm. When the entire immobilized antibody was interacted, there was no additional, non-specific binding observed, which suggested the biointeraction was very specific as expected and independent of the ligand density. No significant steric hindrance was observed at 17.6 nm/$mm^2$ immobilization density. The relationship between the HBsAg concentration in the sample solution and the antigen bound to the chip surface was linear up to ca. $40\mu\textrm{g}$/mL, which is much wider than that of the ELISA method. It appeared the antigen-antibody binding was increased as the immobilized ligand density increased, but verification is warranted. This study showed the potential of this biosensor-based method as a rapid, simple, multi-sample, on-line assay. Once properly validated, it can serve as a more powerful method for HBsAg quantification replacing the current ELISA method.

Effects of Aronia melanocarpa and Korean Red Ginseng Ethanol Extracts Combination on Cytotoxicity induced by Fludarabine, a DNA Chain Terminating Anti-Cancer Drug (DNA 사슬 종결형 항암제인 플루다라빈에 의해 유도된 세포독성에 대한 아로니아-홍삼 에탄올 혼합 추출물의 효과)

  • Kim, Min Seob;Chung, You Heon;Oh, Hong Geun;Park, Jong Kun
    • The Korean Journal of Food And Nutrition
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    • v.30 no.4
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    • pp.673-680
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    • 2017
  • Fludarabine, a chain terminating anti-cancer drug, is a purine analogue that causes DNA strand breaks in normal cells. In this study, we determined if A. melanocarpa and Korean red ginseng extract mixture reduce cytotoxicity of fludarabine. Treatment of HaCaT cells with $10{\mu}M$ of fludarabine for 24 hours decreased cell viability and increased DNA strand breaks. Treatment of A. melanocarpa and Korean red ginseng extract mixture for 24 hours increased cell viability as compared with single extract treatment. The protective effect of these extracts on cell activity increased in a concentration-dependent manner. DNA strand breaks induced by fludarabine decreased as concentration of extract mixture increased. p-H2AX level, a marker of DNA strand breakage, decreased depending on the concentration of extract mixture. The effect of mixed extract of A. melanocarpa and Korean red ginseng on DNA damage is due to the anti-oxidative effect of A. melanocarpa and signal transmission through glucocorticoid receptor upon binding of saponin of Korean red ginseng.

Qualitative Analysis and Plasma Characteristics of Soil from a Desert Area using LIBS Technique

  • Farooq, W. Aslam;Tawfik, Walid;Al-Mutairi, Fahad N.;Alahmed, Zeyad A.
    • Journal of the Optical Society of Korea
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    • v.17 no.6
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    • pp.548-558
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    • 2013
  • In this work, laser induced breakdown spectroscopy (LIBS) is used to investigate soil samples collected from different desert areas of Riyadh city in Saudi Arabia. Both qualitative analysis and plasma parameters are studied via the observed LIBS spectra. These experiments have been done using a Spectrolaser-7000 system with 50 mJ fundamental wavelength of Nd:YAG laser and detection delay time of 1 microsecond. Many spectral lines are highly resolved for many elements like Al, Fe, Mg, Si, Mn, Na, Ca and K. The electron temperatures Te and electron densities Ne, for the constituent of generated LIBS plasma, are determined for all the collected samples. It is found that both Te and Ne vary from one desert area to other. This variation is due to the change of the elemental concentration in different desert areas that affects the sample's matrices. Time dependent measurements have also been performed on the soil samples. While the signal-to-base ratio (SBR) reached its optimal value at 1 microsecond, the plasma parameters Ne and Te reach values of $4{\times}10^{17}cm^{-3}$ and 9235 K, respectively, at 2.5 microsecond. The later indicate that the plasma cooling processes are slow in comparison to the previously observed results for metallic samples. The observed results show also that in the future it is possible to enhance the exploitation of LIBS in the remote on-line environmental monitoring application, by following up only the values of Ne and Te for one element of the soil desert sample using an optical fiber probe.

Neuronal Mechanisms that Regulate Vitellogenesis in the Fruit Fly (노랑초파리 난황형성과정 제어 신경 메커니즘)

  • Kim, Young-Joon;Zhang, Chen
    • Korean journal of applied entomology
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    • v.61 no.1
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    • pp.109-115
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    • 2022
  • Vitellogenesis is the process by which yolk accumulates in developing oocytes. The initiation of vitellogenesis represents an important control point in oogenesis. When females of the model insect Drosophila melanogaster molt to become adults, their ovaries lack mature vitellogenic oocytes, only producing them after reproductive maturation. After maturation, vitellogenesis stops until a mating signal re-activates it. Juvenile hormone (JH) from the endocrine organ known as the corpora allata (CA) is the major insect gonadotropin that stimulates vitellogenesis, and the seminal protein sex peptide (SP) has long been implicated as a mating signal that stimulates JH biosynthesis. In this review, we discuss our new findings that explain how the nervous system gates JH biosynthesis and vitellogenesis associated with reproductive maturation and the SP-induced post-mating response. Mated females exhibit diurnal rhythmicity in oogenesis. A subset of brain circadian pacemaker neurons produce Allatostatin C (AstC) to generate a circadian oogenesis rhythm by indirectly regulating JH and vitellogenesis through the brain insulin-producing cells. We also discuss genetic evidence that supports this model and future research directions.

A study on the implementation of Medical Telemetry systems using wireless public data network (무선공중망을 이용한 의료 정보 데이터 원격 모니터링 시스템에 관한 연구)

  • 이택규;김영길
    • Proceedings of the Korean Institute of Information and Commucation Sciences Conference
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    • 2000.10a
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    • pp.278-283
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    • 2000
  • As information communication technology developed we could check our blood pressure, pulsation electrocardiogram, SpO2 and blood test easily at home. To check our health at ordinary times is able though interlocking the house medical instrument with the wireless public data network This service will help the inconvenience to visit the hospital everytime and will save the individual's time and cost. In each house an organism data which is detected from the human body will be transmitted to the distance hospital and will be essentially applied through wireless public data network The medical information transmit system is utilized by wireless close range network It would transmit the obtained organism signal wirelessly from the personal device to the main center system in the hospital. Remote telemetry system is embodied by utilizing wireless media access protocol. The protocol is embodied by grafting CSMA/CA(Carrier Sense Multiple Access with Collision Avoidance) protocol falling mode which is standards from IEEE 802.11. Among the house care telemetry system which could measure blood pressure, pulsation, electrocardiogram, SpO2 the study embodies the ECC(electrocardiograph) measure part. It within the ECC function into the movable device and add 900㎒ band wireless public data interface. Then the aged, the patients even anyone in the house could obtain ECG and keep, record the data. It would be essential to control those who had a health-examination heart diseases or more complicated heart diseases and to observe the latent heart disease patient continuously. To embody the medical information transmit system which is based on wireless network. It would transmit the ECG data among the organism signal data which would be utilized by wireless network modem and NCL(Native Control Language) protocol to contact through wireless network Through the SCR(Standard Context Routing) protocol in the network it will be connected to the wired host computer. The computer will check the recorded individual information and the obtained ECC data then send the correspond examination to the movable device. The study suggests the medical transmit system model utilized by the wireless public data network.

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Cellular Localization and Translocation of Duplication and Alternative Splicing Variants of Olive Flounder Phospholipase C-δ1 (넙치 3가지 타입 인지질가수분해효소(PLC-δ1)의 세포 내 위치 및 이동)

  • Kim, Na Young;Kim, Moo-Sang;Jung, Sung Hee;Kim, Myoung Sug;Cho, Mi Young;Chung, oon Ki;Ahn, Sang Jung
    • Journal of Life Science
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    • v.27 no.11
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    • pp.1369-1375
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    • 2017
  • The purpose of this study was to investigate the cellular characterization of phospholipase C-${\delta}1$ in olive flounders (Paralichthys olivaceus). In general, phospholipase C signaling pathways are distributed in nuclei at plasma membranes and in cytoplasms, although the pathways' nuclear localization mechanisms are unclear. P. olivaceus duplicates type-A PoPLC-${\delta}1$ (PoPLC-${\delta}1A$), which has a high similarity to the human isoform PLC-${\delta}$; type-B PoPLC-${\delta}1$ (PoPLC-${\delta}1B$ [Sf]), which has a low similarity to the human isoform PLC-${\delta}$ and the alternative splice variant PoPLC-${\delta}1B$ (Lf), which has a nuclear localization signal (NLS) and a nuclear export signal (NES) for nuclear imports and exports, respectively. This study confirmed the effects of the cellular localization and translocation of GFP-tagged PoPLC-${\delta}1A$, PoPLC-${\delta}1B$ (Sf) and PoPLC-${\delta}1B$ (Lf). It administered treatments of $Ca^{2+}$ ionophore ionomycin and endoplasmic reticulum (ER)-$Ca^{2+}$ pump inhibitor thapsigargin to hirame natural-embryo (HINAE) cells. A laser-scanning confocal microscope was used. GFP-tagged PoPLC-${\delta}1A$ was distributed to the cellular organelles, rather than to the cytoplasms and cytomembranes, when PoPLC-${\delta}1B$ (Lf) and PoPLC-${\delta}1B$ (Sf) were localized at the plasma membranes. The treatments of ionomycin and thapsigargin showed the accumulation of PoPLC-${\delta}1A$ in the nuclei when PoPLC-${\delta}1B$ (Lf) nucleocytoplasmic shuttling and PoPLC-${\delta}1B$ (Sf) nucleocytoplasmic shuttling were not observed. The results were the first evidence that PoPLC-${\delta}1A$, which contains functional, intact NES sequences, has a main role in nucleocytoplasmic shuttling and translocation in fish.

A Study on Antitumor Effect and Mechanism of Cortex ulmi pumilae Water Extract on HepG2 Hepatoma cell (유근피(楡根皮) 추출액(抽出液)이 HeoG2 간암세포(肝癌細胞)에 미치는 항암효과(抗癌效果) 및 기전(機轉)에 대(對)한 연구(硏究))

  • Choi, Su-Deock;Park, Young-Kweon;Kim, Gang-San;Kang, Byung-Ki;Han, Sang-Il
    • The Journal of Internal Korean Medicine
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    • v.21 no.2
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    • pp.259-266
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    • 2000
  • Objectives : The effects of aqueous extracts of Cortex ulmi pumilae (a traditional medicine for cancer treatment in oriental medicine) on the induction of apoptotic cell death were investigated in human liver origm hepatoma cell lines, HepG2. Methods : The death of HepG2 cells was markedly induced by the addition of extracts of Cortex ulmi pumilae in a dose-dependent manner. The apoptotic characteristic ladder pattern of DNA strand break was not observed in cell death of HepG2. In addition, it was not shown nucleus chromatin condensation and fragmentation under hoechst staining. However, by the using annexin V staining assay, externalizations of phosphatidylserine in HepG2 cell which were treated with Cortex ulmi pumilae extracts were detected in the early time (at 9 hr after extract treatment). Furthermore, LDH release was not detected in this early stage. Therefore, Cortex ulmi pumilae extracts-induced cell death of HepG2 cells is mediated by apoptotic death signal processes. Result : The activity of caspase 3-like proteases remained in a basal level in HepG2 cells which treated with the extract of Cordyceps sinensis. However, it was markedly increased in HepG2 cells which treated with two extracts of Cortex ulmi pumilae (C.U.P.-C, C.U.P.-K) which were differently extracted (respectively, 2.3 and 3.3 fold). On a while, the phosphotransferase activities of JNK1 was markedly induced in HepG2 cells which were treated with two extracts of Cortex ulmi pumilae. On the contrary, the activation of transcriptional activator, activating protein1(AP-1) and NF-kB were severely decreased by these two extracts of Cortex ulmi pumilae (C.U.P.-C, C.U.P.-K). In addition, antioxidants (GSH and NAC) and intracellular $Ca2^+$ level regulator (Bapta/AM and Thapsigargin) did not affect Cortex ulmi pumilae extracts-induced apoptotic death of HepG2 cells. Conclusions : In conclusion, our results suggest that two extracts of Cortex ulmi pumilae (C.U.P.-C, C.U.P.-K) induces the apoptotic death of human liver origin hepatoma HepG2 cells via activation of caspase 3-like proteases as well as JNK1, and inhibition of transcriptional activators, AP-1 and $NK-{\kappa}B$.

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A Study on the Inhibition of Skin Pigmentation by Lobaric Acid as Protease Activated Receptor-2 Antagonist (Protease Activated Receptor-2의 길항제로서 Lobaric Acid의 피부 색소침착 억제 효능 연구)

  • Goo, Jung Hyun;Lee, Ji Eun;Myung, Cheol Hwan;Park, Jong Il;Hwang, Jae Sung
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.41 no.3
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    • pp.243-252
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    • 2015
  • Melanosome, the pigment granule in melanocyte, determines the color of skin when it moves into the keratinocyte. Inhibition of melanosome transfer from melanocyte to keratinocyte results in skin depigmentation. Protease activated receptor-2 (PAR-2) is involved in signal transduction systems via cell membrane and increases the melasome transfer when it is activated by cleavage of their extracellular amino acid sequence by trypsin or by a peptide such as SLIGKV. Here, we showed that lobaric acid inhibited PAR-2 activation and affected the mobilization of $Ca2^+$. The uptake of fluorescent microspheres and isolated melanosomes from melan-a melanocytes to keratinocytes induced by SLIGKV were inhibited by lobaric acid. Also, confocal microscopy studies illustrated a decreased melanosome transfer to keratinocytes in melanocyte-keratinocyte co-culture system by lobaric acid. In addition, lobaric acid induced visible skin lightening effect in human skin tissue culture model, melanoderm$^{(R)}$. Our data suggest that lobaric acid could be an effective skin lightening agent that works via regulation of phagocytic activity of keratinocytes.