• Title/Summary/Keyword: membrane processes

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Ultrastructures of Oocyte Development and Electrophoretic Patterns of the Yolk Protein Following HCG Treatment in Korean Native Catfish (Silurus asotus)

  • Yoon, J.M.;Chung, E.Y.;Kim, G.W.
    • Asian-Australasian Journal of Animal Sciences
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    • v.14 no.2
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    • pp.174-183
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    • 2001
  • During the rapid phase of gonadal development of the freshwater teleost, the catfish (Silurus asotus), the influence of hCG upon the inducement of final oocyte maturation and spawning was investigated electrophoretically and ultrastructurally. The electrophoretic patterns obtained were different in the presence and absence of some of the major or minor zones, because of the hormone level in catfish. The vitellogenin of hormone-treated fish was stained more intensively than that of sham-treated fish. These proteins showed some minor or main bands of egg extracts which migrated at positions corresponding to molecular weights of approximately 90,000. However, the thickness of electrophoretic band in molecular weight for hCG-treated fish was slightly lower than that for saline control. It seemed the plasma protein with molecular weight of approximately 45,000 in hCG-treated fish disappeared. In contrast to the control fish, the ovaries in the catfish treated with hCG shows a marked ultrastructural change under the electron microscope. No dilated profiles were seen in the granulosa cells of the mature oocyte before ovulation. After germinal vesicle breakdown (GVBD), the zona radiata interna (ZRI) becomes more compact, and there is a loss of all the processes from the pore canals. There is a wide space between the vitelline membrane and zona radiata. Also, during final maturation, the microvillar processes from the oocyte are seen no longer to penetrate deeply into the extracellular spaces of the overlying granulosa cells, and the reticulate patterns of the zona radiata interna becomes occluded, giving the zona radiata a more solid appearance. It has been possible to initiate 100% oocyte maturation in yolk granules and follicles in vivo by treatment with hCG and a high water temperature ($27^{\circ}C$). In hCG-treated fish, the percentages of successful artificial fertilization and hatching were maximal at 15 h after a single injection. It seems clear that a long acting preparation containing hCG can be successfully used in prespawning fish to advance the final events of gonadal maturation and initiate spawning. Further studies are necessary to evaluate the potential of hCG to either stimulate or inhibit the reproductive development of fish at other stages of the seasonal reproductive cycle.

Visual Cells in the Retina of Iksookimia longicorpa (Pisces; Cobitidae) of Korea (한국산 미꾸리과 어류 왕종개 Iksookimia longicorpa 망막의 시각세포)

  • Kim, Jae Goo;Park, Jong Young
    • Korean Journal of Ichthyology
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    • v.27 no.4
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    • pp.257-262
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    • 2015
  • The visual cells in the retina of Iksookimia longicorpa (Pisces, Cobitidae) were investigated by light and scanning electron microscopes. The retina ($216.42{\pm}13.36{\mu}m$) has several layers, and the visual cell layer consists of unequal double cones and large rods. In a double cone, two members are unequal such that one cone is longer than the other (long element $26.42{\pm}1.7{\mu}m$, short element $16.82{\pm}1.1{\mu}m$). The cones form a row mosaic pattern in which the partners of double cones are linearly oriented with a large rod. The visual cells observed have an outer segment (hematophilic), inner segment (eosinophilic). In scanning electron microscopy, the outer segment links to inner segment by so-called calyceal piles (calyceal processes) of membrane discs surrounded by double membranes.

The Ultrastructure of the Cutaneous Pigment Cells in the Amphibia (양서류 피부 색소세포의 미세구조)

  • 김한화;노용태;지영득;문영화
    • The Korean Journal of Zoology
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    • v.24 no.3
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    • pp.133-144
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    • 1981
  • The ultrastructures of the pigment cells in the Asiatic land salamander (Hynobius leechi) dorsal skin were obtained by means of electron microscope. The results were as follows; 1. The pigment cells of the epidermis consisted of the melanocytes in the germinal layer and of the melanophores distributing to the keratinocyte layer. The traits of these cells in the epidermis were as follows: A. The nuclei of the melanocytes were round or oval in shape and appeared as partly small or large infoldings of the nuclear envelope. B. Rough-surfaced endoplasmic reticulums and Golgi complexes were well developed in infranuclear cytoplasm. Many ribosomes were mainly distributed around the perinuclear portion. C. The melanosomes of the melanocytes were observed as a found or an oval shape and strong electron-dense or less electron-dense melanosomes were observed. D. The infoldings of the nuclear envelope in the melanophore were partly found deeper than those of the melanocyte. The cytoplasm of the melanophore filled with melanosomes caused organelles not to be observed in that. 2. The pigment cells in the dermis were composed of the xanthophores just beneath basement membrane and the melanophores in the connective tissue. The traits of these cells in the dermis were as follows: A. The xanthophores contained round or oval vesicles, and these vesicles were divided into 6 types (type I pterinosome, type II pterinosome, type III pterinosomes, type iv pterinosome, type V pterinosome, type VI pterinosome). B. Most of the nuclei of the melanophores in the dermis were elongate in shape, and a portion of the nuclear envelope was deep infolded. C. Becuase the cytoplasm was filled with the melanosomes of the same electron-density, organelles were not observed in the cytoplasm. D. Two processes of the melanophore in the dermis extended in parallel with a xanthophore and the cytoplasm in those processes were filled with the melanosomes.

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Time-based Expression Networks of Genes Related to Cold Stress in Brassica rapa ssp. pekinensis (배추의 저온 스트레스 처리 시간대별 발현 유전자 네트워크 분석)

  • Lee, Gi-Ho;Yu, Jae-Gyeong;Park, Young-Doo
    • Horticultural Science & Technology
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    • v.33 no.1
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    • pp.114-123
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    • 2015
  • Plants can respond and adapt to cold stress through regulation of gene expression in various biochemical and physiological processes. Cold stress triggers decreased rates of metabolism, modification of cell walls, and loss of membrane function. Hence, this study was conducted to construct coexpression networks for time-based expression pattern analysis of genes related to cold stress in Chinese cabbage (Brassica rapa ssp. pekinensis). B. rapa cold stress networks were constructed with 2,030 nodes, 20,235 edges, and 34 connected components. The analysis suggests that similar genes responding to cold stress may also regulate development of Chinese cabbage. Using this network model, it is surmised that cold tolerance is strongly related to activation of chitinase antifreeze proteins by WRKY transcription factors and salicylic acid signaling, and to regulation of stomatal movement and starch metabolic processes for systemic acquired resistance in Chinese cabbage. Moreover, within 48 h, cold stress triggered transition from vegetative to reproductive phase and meristematic phase transition. In this study, we demonstrated that this network model could be used to precisely predict the functions of cold resistance genes in Chinese cabbage.

Calpains and Apoptosis

  • Tagliarino, Colleen;Pink, John J.;Boothman, David A.
    • Animal cells and systems
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    • v.5 no.4
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    • pp.267-274
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    • 2001
  • Calpains are a family of cysteine proteases existing primarily in two forms designated by the $Ca^{2+}$ concentration needed for activation in vitro, $\mu$-calpain (calpain-I) and m-calpain (calpain-II). The physiologica1 roles of calpains remain unclear. Many groups have proposed a role for calpains In apoptosis, but their patterns of activation are not well characterized. Calpains have been implicated in neutrophil apoptosis, glucocorticoid-induced thymocyte apoptosis, as well as many other apoptotic pathways. Calpain activation in apoptosis is usually linked upstream or downstream to caspase activation, or in a parallel pathway alongside caspase activation. Calpains have been suggested to be involved in DNA fragmentation (via endonuclease activation), but also as effector proteases that cleave cellular proteins involved in DNA repair, membrane associated proteins and other homeostatic regulatory proteins. Recently, our laboratory demonstrated $\mu$-calpain activation in NAD(P)H: quinone oxidoreducatse 1 (NQO1)-expressing cells after exposure to $\beta$-lapachone, a novel quinone and potential chemo- and radio-therapeutic agent. Increased cytosolic $Ca^{2+}$ in NQO1-expressing cells after $\beta$-lapachone exposures were shown to lead to $\mu$-calpain activation. In turn, $\mu$-calpain activation was important for substrate proteolysis and DNA fragmentation associated with apoptosis. Upon activation, $\mu$-calpain translocated to the nucleus where it could proteolytically cleave PARP and p53. We provided evidence that $\beta$-lapachone-induced, $\mu$-calpain stimulated, apoptosis did not involve any of the known caspases; known apoptotic caspases were not activated after $\beta$-lapachone treatment of NQO1-expressing cells, nor did caspase inhibitors have any effect on $\beta$-1apachone-induced cell death. Elucidation of processes by which $\beta$-1apachone-stimulated $\mu$-calpain activation and calpains ability to activate endonucleases and induce apoptosis independent of caspase activity will be needed to further develop/modulate $\beta$-lapachone for treatment of human cancers that over-express NQO1.

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Dynamics of Nanopore on the Apex of the Pyramid

  • Choi, Seong-Soo;Yamaguchi, Tokuro;Park, Myoung-Jin;Kim, Sung-In;Kim, Kyung-Jin;Kim, Kun-Ho
    • Proceedings of the Korean Vacuum Society Conference
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    • 2012.08a
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    • pp.187-187
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    • 2012
  • In this report, the plasmonic nanopores of less than 5 nm diameter were fabricated on the apex of the pyramidal cavity array. The metallic pyramidal pit cavity can also utilized as the plasmonic bioreactor, and the fabricated Au or Al metallic nanopore can provide the controllable translocation speed down using the plasmonic optical force. Initially, the SiO2 nanopore on the pyramidal pit cavity were fabricated using conventional microfabrication techniques. Then, the metallic thin film was sputter-deposited, followed by surface modification of the nanometer thick membrane using FESEM, TEM and EPMA. The huge electron intensity of FESEM with ~microsecond scan speed can provide the rapid solid phase surface transformation. However, the moderate electron beam intensity from the normal TEM without high speed scanning can only provide the liquid phase surface modification. After metal deposition, the 100 nm diameter aperture using FIB beam drilling was obtained in order to obtain the uniform nano-aperture. Then, the nanometer size aperture was reduced down to ~50 nm using electron beam surface modification using high speed scanning FESEM. The followed EPMA electron beam exposure without high speed scanning presents the reduction of the nanosize aperture down to 10 nm. During these processes, the widening or the shrinking of the nanometer pore was observed depending upon the electron beam intensity. Finally, using 200 keV TEM, the diameter of the nanopore was successively down from 10 nm down to 1.5 nm.

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Physiological Changes of Saccharomyces cerevisiae by High Voltage Pulsed Electric Field Treatments (고전압 펄스 전기장 처리에 의한 Saccharomyces cerevisiae의 생리적 변화)

  • Park, Hee Ran;Yoon, So Jung;Park, Han-Sul;Shin, Jung-Kue
    • Korean Journal of Food Science and Technology
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    • v.45 no.5
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    • pp.590-597
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    • 2013
  • High voltage pulsed electric fields (PEF) treatment is a promising non-thermal processing technology that can replace or partially substitute for thermal processes. The aim of this research was to investigate the microbial inactivation mechanisms by PEF treatment in terms of physiological changes to Saccharomyces cerevisiae. PEF was applied at the electric field strength of 50 kV/cm, treatment time of 56 ${\mu}s$ and temperature of $40^{\circ}C$. The microbial cells treated with PEF showed loss of salt tolerance on the cell membrane and collapse of the relative pH gradient on in-out of cells. Cell death or injury resulted from the breakdown of homeostasis, decreased $H^+$-ATPase activity, and loss of glycolysis activity.

Inducing Effects of Rubus coreanus on Cell Death and Apoptotic Gene Expressions in Human Breast Cancer Cells (복분자의 유방암 세포 사멸 및 사멸 유전자 발현 유도 효과)

  • Kim, Hee-Jung;Kang, Keum-Jee
    • Journal of the East Asian Society of Dietary Life
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    • v.23 no.6
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    • pp.723-732
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    • 2013
  • We investigated the inducing effects of Rubus coreanus extract (RCE) on apoptosis and its related gene expressions in human breast cancer cells. MDA-MB-231 cells were cultured in the presence of 0, 200, 300, and $400{\mu}g/mL$ RCE for 24h. MTT assay demonstrated that relative cell viability measured a decrease in a dose-dependent manner (p<0.05). This dependency was also found in the increasing levels of cell death by a dual staining with Hoechst 33322 and propidium iodide (p<0.05). These close associations was also observed by different stages of apoptotic processes, as shown by an Apoptosis Detection Kit. To determine whether the alterations in such cell activities obtained above cause the induction of apoptotic genes, PT-PCR was performed expressions of both Bcl-2 and Bax mRNAs. The Bcl-2/Bax ratio which is an important indicator of apoptosis, was found to have significantly decreased dose dependence (p<0.05). Western blot analysis also demonstrated that Caspase-3 significantly increases in a dose-dependent manner (p<0.05) in addition to similar alterations of other proteins examined. Taken these results together, the ethanolic RCE used induces a reduction in cell viability along with increased membrane permeability. This leads to a precautious apoptotic process and, subsequently, cell death through the apoptotic pathway involving Bax and Caspase-3 in human breast cancer MDA-MB-231 cells.

Aqueous U(VI) removal by green rust and vivianite at phosphate-rich environment

  • Sihn, Youngho;Yoon, In-Ho
    • Membrane and Water Treatment
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    • v.11 no.3
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    • pp.207-215
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    • 2020
  • Vivianite (Fe32+(PO4)2·8H2O) and green rust ([Fe42+Fe23+(OH)-12][SO42-·2H2O]2-), ferrous containing minerals, could remove aqueous U(VI) in 5 min. and the efficiencies of green rust were roughly 2 times higher than that of vivianite. The zeta potential measurement results implies that the better performance of green rust might be attributed to the favorable surface charge toward uranyl phosphate species. The removal behaviors of the minerals were well fitted by pseudo-second order kinetic model (R2 > 0.990) indicating the dominant removal process was chemical adsorption. Effects of Ca2+ and CO32- at pH 7 were examined in terms of removal kinetic and capacity. The kinetic constants of aqueous U(VI) were 8 and 13 times lower (0.492 × 10-3 g/(mg·min); 0.305 × 10-3 g/(mg·min)) compared to the value in the absence of the ions. The thermodynamic equilibrium calculation showed that the stable uranyl species (uranyl tri-carbonate) were newly formed at the condition. Surface investigation on the reacted mineral with uranyl phosphates species were carried out by XPS. Ferrous iron and U(VI) on the green rust surface were completely oxidized and reduced into Fe(III) and U(IV) after 7 d. It suggests that the ferrous minerals can retard U(VI) migration in phosphate-rich groundwater through the adsorption and subsequent reduction processes.

Cromakalim Blocks Membrane Phosphoinositide Activated Signals in the Guinea Pig Lung Mast Cells Stimulated with Antigen-Antibody Reactions

  • Ro, Jai-Youl;Kim, Ji-Young;Kim, Kyung-Hwan
    • The Korean Journal of Physiology and Pharmacology
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    • v.2 no.2
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    • pp.251-260
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    • 1998
  • Cromakalim (BRL 34915), known as an airway smooth muscle relaxant, inhibited the releases of mediators in the antigen-induced mast cell activation. It has been suggested that cromakalim, in part, inhibited mediator releases by inhibiting the initial increase of 1,2-diacylglycerol (DAG) produced by the activation of the other phospholipase system which is different from phosphatidylcholine-phospholipase D pathway. The aim of this study is to further examine the inhibitory mechanism of cromakalim on the mediator release in the mast cell activation. Guinea pig lung mast cells were purified by using enzyme digestion and percoll density gradient. In purified mast cells prelabeled with $[^3H]PIP_2$, phospholipase C (PLC) activity was assessed by the production of $[^3H]$insitol phosphates. Protein kinase C (PKC) activity was assessed by measuring the protein phosphorylated from mast cells prelabeled with $[{\gamma}-32P]ATP$, and Phospholipase $A_2\;(PLA_2)$ activity by measuring the lyso-phosphatidylcholine produced from mast cell prelabeled with 1-palmitoyl-2-arachidonyl $phosphatidyl-[^{14}C]choline$. Histamine was assayed by fluorometric analyzer, and leukotrienes by radioimmunoassay. The PLC activity was increased by activation of the passively sensitized mast cells. This increased PLC activity was decreased by cromakalim pretreatment. The PKC activity increased by the activation of the passively sensitized mast cells was decreased by calphostin C, staurosporine and cromakalim, respectively. The $PLA_2$ activity was increased in the activated mast cells. The pretreatment of cromakalim did not significantly decrease $PLA_2$ activity. These data show that cromakalim inhibits histamine release by continuously inhibiting signal transduction processes which is mediated via PLC pathway during mast cell activation, but that cromakalim does not affect $PLA_2$ activity related to leukotriene release.

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