• Title/Summary/Keyword: flow cell

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Characterization of Monoclonal Antibodies against Human Leukocyte Common Antigen (CD45)

  • Shin, Hyang-Mi;Cho, Woon-Dong;Lee, Geon-Kook;Lee, Seon-Hwa;Lee, Kyung-Mee;Ji, Gil-Yong;Yoon, Sang-Soon;Koo, Ji-Hae;Lee, Ho-Chang;Lee, Ki-Hyeong;Song, Hyung-Geun
    • IMMUNE NETWORK
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    • v.11 no.2
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    • pp.114-122
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    • 2011
  • Background: The leukocyte common antigen (CD45) is a transmembrane-type protein tyrosine phosphatase that has five isoforms. Methods: We generated seven murine mAbs against human CD45 by injecting cells from different origins, such as human thymocytes, PBMCs, and leukemic cell lines. By using various immunological methods including flow cytometry, immunohistochemistry, and immunoprecipitation, we evaluated the reactivity of those mAbs to CD45 of thymus as well as tonsil lysates. Furthermore, we transiently transfected COS-7 cells with each of gene constructs that express five human CD45 isoforms respectively, and examined the specificities of the mAbs against the transfected isoforms. Results: In case of thymocytes, lymphocytes, and monocytes, all the seven mAbs demonstrated positive reactivities whereas none was reactive to erythrocytes and platelets. The majority of immune cells in formalin-fixed paraffin-embedded thymus and tonsil tissues displayed strong membranous immunoreactivity, and the main antigen was detected near 220 kDa in all cases. Among the mAbs, four mAbs (AP4, DN11, SHL-1, and P6) recognized a region commonly present in all the five isoforms. One mAb, YG27, recognized four isoforms (ABC, AB, BC, and O). Two mAbs, P1 and P14, recognized the isoforms that contain exon A encoded regions (ABC and AB). Conclusion: In this study, we confirmed that AP4, DN11, SHL-1, YG27 and P6, are mAbs reactive with the CD45 antigen whereas P1 and P14 are reactive with the CD45RA antigen.

Fouling Study with Binary Protein Mixtures in Microfilration (이성분계 단백질 혼합물의 미세막 분리공정에서 막오염에 관한 연구)

  • Ahn, Byung Hun;Moon, Dong Ju;Yoo, Kye Sang;Ho, Chia Chi
    • Korean Chemical Engineering Research
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    • v.43 no.2
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    • pp.236-242
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    • 2005
  • Membrane fouling by protein mixtures during microfiltration has been investigated for binary mixtures of bovine serum albumin (BSA), casein, lysozyme, pepsin, and ovalbumin. Filtration experiments were carried out using $0.2{\mu}m$ polycarbonate track-etched (PCTE) membrane in a stirred cell under constant transmembrane pressure (14 kPa) and concentration of hydrogen ion (pH=11) to study the effect of mixture composition on filtrate flux decline. Flux decline data were analyzed using a pore blockage-cake formation model developed recently. It was found that the model is in a good agreement with the experimental data. Fouling parameters such as the rate of pore blockage(${\alpha}$), the initial resistance of the protein deposit ($R_{po}$) and the increasing rate of the protein layer resistance(${\beta}$) were used to evaluate the rate of filtrate flow by membrane fouling in the binary mixture system. Generally, the trend of ${\alpha}$ is comparable with that of filtrate flux decline. It was also found that fast flux decreasing was observed over the binary mixture containing casein. The result is due to high value of the initial resistance of the protein deposit ($R_{po}$) over casein.

Crystal Structure of a Bromine Sorption Complex of Dehydrated Calcium and silver Exchanged Zeolite A (칼슘 및 은 이온으로 치환된 제올라이트 A를 탈수한 후 브롬을 흡착한 결정구조)

  • Bae, Myung-Nam;Kim, Un-Sik;Kim, Yang
    • Korean Journal of Crystallography
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    • v.8 no.2
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    • pp.127-131
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    • 1997
  • The crystal structure of a bromine sorption complex of vacuum-dehydrated Ag+ and Ca2+ exchanged zeolite A(a=12,234(1) Å) has been determined by single-crystal X-ray diffraction methods in the cubic space group Pm3m. The crystal was prepared by flow method using exchange solution in which mole ratio of AgNo3 and Ca(NO3)2 was 1:150 with a total concentration of 0.05M. The crystal was dehydrated at 360℃ and 2 ×10-6 Torr for 2days, followed by exposure to 180 Torr of Br2 vapor for 20min. full-matrix least-squares refinements converged to the final error indices of R1=0.111 and R2=0.101 using 90 reflections for which I>3o(I). About 3.1 Ag+ ions and 4.45 Ca2+ ions lie on the two crystallographically nonequivalent three-fold axes associated with 6-ring oxygens. A total of six bromine molecules are sorbed per unit cell. Each bromine molecule approaches a framework oxide ions axially (Br-Br-O=171(2)', O-Br=3.25(6) Å; and Br-Br=2,61(8) Å by a charge-transfer interaction.

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The role of methylenetetrahydrofolate reductase C677T polymorphism on the peripheral blood natural killer cell proportion in women with unexplained recurrent miscarriages

  • Park, Chan-Woo;Han, Ae-Ra;Kim, Joanne-Kwak;Park, So-Yeon;Han, Jung-Yeol;Koong, Mi-Kyoung;Song, In-Ok;Yang, Kwang-Moon
    • Clinical and Experimental Reproductive Medicine
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    • v.38 no.3
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    • pp.168-173
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    • 2011
  • Objective: To examine the association between methylenetetrahydrofolate reductase (MTHFR) C677T polymorphism and hyperhomocysteinemia in women with unexplained recurrent miscarriages (RM) and to investigate the association between MTHFR genotype variants and alloimmune activation, proportion of peripheral blood natural killer (pbNK) cells. Methods: A total of 39 patients with a history of two or more unexplained miscarriages were recruited to this study. The controls were women who had a live birth without a history of RM (n=50). The proportion of pbNK cells was measured by flow cytometry. Plasma homocysteine levels and the incidence of the MTHFR variant of the RM and control groups were compared. The proportion of pbNK cells was compared to the MTHFR variants in the RM group. Results: No differences were found between the two groups' mean plasma homocysteine levels ($7.6{\pm}1.5{\mu}mol$/L vs. $7.1{\pm}2.1{\mu}mol$/L) or incidence of the MTHFR genotype variant (CC, 35% vs. 33%; CT, 40% vs. 53%; and TT, 25% vs. 14%). In the RM group, individuals with the TT variant ($7.7{\pm}1.1{\mu}mol$/L) had higher homocysteine levels than those with the CC and CT variants ($7.4{\pm}1.9{\mu}mol$/L and $7.4{\pm}1.2{\mu}mol$/L) and those with the CT variant ($19.2{\pm}8.1%$) had a higher proportion of CD3-/CD56+ pbNK cells than those with the CC and TT variants ($17.7{\pm}6.6%$ and $17.9{\pm}7.0%$), but the results of both comparisons were statistically insignificant. Conclusion: These preliminary results show no difference in plasma homocysteine levels between the RM and control groups or among MTHFR genotype variants in the RM group, which may suggest that the plasma homocysteine level is difficult to use as a predictive marker of RM in the Korean population. A study of a larger number of patients is needed.

Role of Trophobolast in Implantation and Placenta Development (착상 및 태반 발달과정에 따른 영양막세포의 역할)

  • Kim, Gi-Jin
    • Clinical and Experimental Reproductive Medicine
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    • v.37 no.3
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    • pp.181-189
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    • 2010
  • The placenta, which is a temporary organ derived from the fetus during pregnancy, is critical to support fetus development via optimal regulation between mother and fetus. Trophoblast as a major cell population of the placenta is one of the earliest to differentiate and shows an extensive proliferation or/and differentiation up to the formation of the placenta. The role of the trophoblast show dynamic changes from early embryo implantation to placentation during pregnancy. Implantation of the blastocyst into the endometrium of the maternal uterus is mediated by invasion of the differentiated trophoblast (e.g. syncytiotrophoblast) from the trophectoderm. During pregnancy, the unique role of the trophoblast is to invasion, eroding, and metastasizing in the placenta as well as to ensure appropriate bidirectional nutrient or waste flow required for growth and maturation of the embryo. The dysfunction of the trophoblast during pregnancy can result in several gynecological diseases including preeclampsia and congenital malformation in neonatal medicine. Therefore, trophoblasts act as a conclusive factor in placental and fetal development. This brief review outlines the classification of trophoblast and its function in the placenta during pregnancy. Also, we introduce the latest research in trophoblast for implantation and the placenta development, and the application potential of trophoblast for infertility and obstetrical diseases.

Production of Mycelia and Water Soluble Polysaccharides from Submerged Culture of Ganoderma applanatum Using Different Types of Bioreactor (생물반응기 유형에 따른 잔나비불로초(Ganoderma applanatum)의 균사체 및 수용성 다당체 생산 특성)

  • Lee, Wi-Young;Park, Young-Ki;Ahn, Jin-Kwon;Park, So-Young
    • The Korean Journal of Mycology
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    • v.34 no.1
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    • pp.1-6
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    • 2006
  • In order to select a suitable bioreactor type for the submerged cultivation of Ganoderma applanatum, both growth characteristics and polysaccharides production were compared among four different types of bioreactor. These include an external-loop type air-lift bioreactor (ETAB), a balloon type air bubble bioreactor (BTBB), a column type air bubble bioreactor (CTBB) and a stirrer type bioreactor (STB). The mycelial biomass produced from the reactors were in decreasing order: ETAB ($7\;g/{\ell}$) > BTBB ($6.2\;g/{\ell}$) > STB ($6\;g/{\ell}$) > CTBB ($5\;g/{\ell}$). Maximal soluble exopolysaccharides ($1\;g/{\ell}$) and endopolysaccharides (2.7%) were also obtained from ETAB. Thus, the ETAB was most suitable for submerged culture of G applanatum mycelium. Based on the results, ETAB was chosen for further detailed study. The most effective aeration rate for the mycelial growth in ETAB ranged from 0.05 to 0.1 vvm. For the maximal production, the mycelium at the initial growth stage needed low aeration rate to reduce cell damages by fluid flow. However, as the mycelia grew, the culture became viscous and thus needed higher aeration. The molecular weight of exopolysaccharides obtained from the culture grown in ETAB was higher than that from the culture grown in other bioreactors.

Induction of Apoptosis by Bee Venom in A549 Human Lung Epithelial Cancer Cells through Modulation of Bcl-2 and IAP Family and Activation of Caspases (Bcl-2 및 IAP family의 발현 변화와 caspase 활성을 통한 봉독의 인체폐암세포 apoptosis 유도)

  • Woo, Hyun-Joo;Kim, Hyun-Joong;Hong, Su-Hyun;Hong, Sang-Hoon;Choi, Byung-Tae;Lee, Yong-Tae;Park, Dong-Il;Choi, Yung-Hyun
    • Journal of Life Science
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    • v.17 no.11
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    • pp.1596-1600
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    • 2007
  • Bee venom is used to treat inflammatory diseases in Korean traditional medicine and has been known to inhibit proliferation and induce apoptosis in cancer cells. However, the molecular mechanisms involved in bee venom-induced apoptosis are still uncharacterized in human lung cancer cells. In the present study, we investigated the effects of bee venom on the apoptosis of A549 human lung epithelial cancer cells. Treatment of bee venom inhibited the cell viability and induced apoptosis in a concentration-dependent manner as measured by hemocytometer counts, fluorescence microscopy and flow cytometry analysis. Bee venom-induced apoptosis in A549 cells was associated with a marked inhibition of anti-apoptotic Bcl-2 expression without significant changes in the levels of Bax and Bcl-xL. Bee venom treatment also inhibited the levels of IAP family members such as cIAP-1 and cIAP-2 and induced the proteolytic activation of caspase-3 and caspase-9. Although further studies are needed, the present results suggest that apoptotic signals evoked by bee vemon in A549 cancer cells may converge caspases activation through a down-regulation of Bcl-2 rather than an up-regulation of Bax. These findings provide important insights into the possible molecular mechanisms of the anti-cancer activity of bee vemon in human cancer cells.

PLZF+ Innate T Cells Support the TGF-β-Dependent Generation of Activated/Memory-Like Regulatory T Cells

  • Kang, Byung Hyun;Park, Hyo Jin;Park, Hi Jung;Lee, Jae-Il;Park, Seong Hoe;Jung, Kyeong Cheon
    • Molecules and Cells
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    • v.39 no.6
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    • pp.468-476
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    • 2016
  • PLZF-expressing invariant natural killer T cells and CD4 T cells are unique subsets of innate T cells. Both are selected via thymocyte-thymocyte interaction, and they contribute to the generation of activated/memory-like CD4 and CD8 T cells in the thymus via the production of IL-4. Here, we investigated whether $PLZF^+$ innate T cells also affect the development and function of $Foxp3^+$ regulatory CD4 T cells. Flow cytometry analysis of the thymus and spleen from both CIITA transgenic C57BL/6 and wild-type BALB/c mice, which have abundant $PLZF^+$ CD4 T cells and invariant natural killer T cells, respectively, revealed that $Foxp3^+$ T cells in these mice exhibited a $CD103^+$ activated/memorylike phenotype. The frequency of $CD103^+$ regulatory T cells was considerably decreased in $PLZF^+$ cell-deficient $CIITA^{Tg}Plzf^{lu/lu}$ and $BALB/c.CD1d^{-/-}$ mice as well as in an IL-4-deficient background, such as in $CIITA^{Tg}IL-4^{-/-}$ and $BALB/c.IL-4^{-/-}$ mice, indicating that the acquisition of an activated/ memory-like phenotype was dependent on $PLZF^+$ innate T cells and IL-4. Using fetal thymic organ culture, we further demonstrated that IL-4 in concert with TGF-${\beta}$ enhanced the acquisition of the activated/memory-like phenotype of regulatory T cells. In functional aspects, the activated/ memory-like phenotype of Treg cells was directly related to their suppressive function; regulatory T cells of $CIITA^{Tg}PIV^{-/-}$ mice more efficiently suppressed ovalbumin-induced allergic airway inflammation compared with their counterparts from wild-type mice. All of these findings suggest that $PLZF^+$ innate T cells also augmented the generation of activated/memory-like regulation via IL-4 production.

Reactive oxygen species-dependent apoptosis induction by water extract of Citrus unshiu peel in MDA-MB-231 human breast carcinoma cells

  • Kim, Min Yeong;Choi, Eun Ok;HwangBo, Hyun;Kwon, Da He;Ahn, Kyu Im;Kim, Hong Jae;Ji, Seon Yeong;Hong, Su-Hyun;Jeong, Jin-Woo;Kim, Gi Young;Park, Cheol;Choi, Yung Hyun
    • Nutrition Research and Practice
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    • v.12 no.2
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    • pp.129-134
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    • 2018
  • BACKGROUND/OBJECTIVES: Although several recent studies have reported the anti-cancer effects of extracts or components of Citrus unshiu peel, which has been used for various purposes in traditional medicine, the molecular mechanisms for their effects remain unclear. In the present study, the anti-cancer activity of a water-soluble extract of C. unshiu peel (WECU) in MDA-MB-231 human breast carcinoma cells at the level of apoptosis induction was investigated. MATERIALS/METHODS: Cytotoxicity was evaluated using the MTT assay. Apoptosis was detected using DAPI staining and flow cytometry analyses. Mitochondrial membrane potential, reactive oxygen species (ROS) assay, caspase activity and Western blotting were used to confirm the basis of apoptosis. RESULTS: The results indicated that WECU-induced apoptosis was related to the activation of caspase-8, and -9, representative initiator caspases of extrinsic and intrinsic apoptosis pathways, respectively, and caspase-3 accompanied by proteolytic degradation of poly(ADP-ribose) polymerase and down-regulation of the inhibitors of apoptosis protein family members. WECU also increased the pro-apoptotic BAX to anti-apoptotic BCL-2 ratio, loss of mitochondrial membrane potential and cytochrome c release from mitochondria to cytoplasm. Furthermore, WECU provoked the generation of ROS, but the reduction of cell viability and induction of apoptosis by WECU were prevented when ROS production was blocked by antioxidant N-acetyl cysteine. CONCLUSIONS: These results suggest that WECU suppressed proliferation of MDA-MB-231 cells by activating extrinsic and intrinsic apoptosis pathways in a ROS-dependent manner.

Numerical Model for Spreading of Cochlodinium Bloom in the Southern Coastal Waters in Korea (한국 남해안에서 Cochlodinium적조 확산모델)

  • Kwon Chul Hui;Cho Ku Dae
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.35 no.6
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    • pp.568-577
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    • 2002
  • The spreading Cocuoainim polykikoides bloom in the southern coastal waters of Korea was simulated using numerical model including the physical processes of water flow and the chemical processes of increasing cell of C. polykikoides by uptake of dissolved nutrients. The circulation of sea water was simulated by two dimensional tide model reflecting the main four tidal components of $M_2,\;S_2,\;K_1,\;O_1$, and permanent current was driven by inflow/outflow across open boundaries. According to the result of model which tidal and permanent current were reflected simultaneously, eastward flows entering the southern waters from the western waters of Korea are dominant but westward flows are weak relatively. These result suggest that it is difficult for initial C. polykikoides bloom generated in the coastal waters of Goheung to move to the western coast of Korea through Jeju Strait. For spreading model of C. poiyhikoides, the range of generating distribution and the generating time of C. polykikoides bloom in coastal area are similar to those of observation data in the field. Wind is the most important factor in moving and distribution of red tide. Permanent current flowing eastward is also considered to be important factor and tidal current was a little influenced.