• Title/Summary/Keyword: membrane treatment

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Induction of Autolysis and Autoplast Formation of Anaerobic Clostridium thermohydrosulfuricum (혐기성 Clostridium thermohydrosulfuricum의 Autolysis 및 Autoplast 형성유도)

  • 김욱한;박동찬;정기택;이용현
    • Korean Journal of Microbiology
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    • v.27 no.4
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    • pp.357-365
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    • 1989
  • Induction conditions for autolysis and autoplast formation of thermophilic Clostridium thermohydrosulfuricum were studied. The cells in the initial exponential growth phase were well autolysed in Tris-HCl buffer or inorganic buffers containing univalents, such as $K^{+}$ and $Na^{+}$ , and chemicals such as cysteine-HCl, sorbitol and glycerol. Meanwhile, autolysis induction was slightly inhibited by divalents, such as $Mg^{2+}, Mn^{2+}, Ca^{2+}, Ni^{2+}$, and strongly by divalents, such as $Fe^{2+}, Cu^{2+}$ and citric acid. The autolysis was stimulated when the cells were grown in the medium containing ampicillin that inhibites cell wall synthesis, meanwhile, it was slightly inhibited by nucleic acids and protein synthesis inhibitors. The optimal pH and temperature for the induction of autolysis were 7.5 and $60^{\circ}C$, respectively. On the other hand, the cells were autoplasted without lysozyme treatment during autolysis due to the stabilization of protoplasmic membrane in the presence of divalents such as $Mg^{2+}, Mn^{2+}, Ca^{2+}, Ni^{2+}$. Autoplast formation was mostly induced at $37^{\circ}C$ in 50mM Tris-HCl buffer (pH 7.5) containing 20 mM $MgCl^{2}$ and 0.3 M glycerol, and in the late exponential growth phase growing cell.

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Role of $K^+$ Channels in $H_2O_2$- and Cryo-induced Apoptosis of Mouse and Bovine Embryos (과산화수소와 동결에 의해 유도된 생쥐와 소 수정란의 사멸에 있어서 칼륨 통로의 역할)

  • Choe, Changyong;Kim, Chang-Woon;Kang, Dawon;Han, Jaehee
    • Journal of Embryo Transfer
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    • v.29 no.3
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    • pp.249-255
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    • 2014
  • Programmed cell death or apoptosis is associated with changes in $K^+$ concentration in many cell types. Recent studies have demonstrated that two-pore domain $K^+$ ($K_{2P}$) channels are involved in mouse embryonic development and apoptotic volume decrease of mammalian cells. In cerebellar granule neurons that normally undergo apoptosis during the early developmental stage, TASK-1 and TASK-3, members of $K_{2P}$ channels, were found to be critical for cell death. This study was performed to identify the role of $K^+$ channels in the $H_2O_2$-induced or cryo-induced cell death of mouse and bovine embryos. Mouse and bovine two-cell stage embryos (2-cells) exposed to $H_2O_2$ for 4 h suffered from apoptosis. The 2-cells showed positive TUNEL staining. Treatment with high concentration of KCl (25mM) inhibited $H_2O_2$-induced apoptosis of 2-cells by 19%. Cryo-induced death in bovine blastocysts showed positive TUNEL staining only in the cells near the plasma membrane. Cryoprotectant supplemented with 25 mM KCl reduced apoptosis slightly compared to cryoprotectant supplemented with 5 mM KCl. However, the combination of antioxidants (${\beta}$-mercaptoethanol) with 25 mM KCl significantly decreased the rate of $H_2O_2$-induced and cryo-induced apoptosis compared to treatments with only antioxidants or 25 mM KCl. These results show that blockage of $K^+$ channel efflux for a short-time reduces $H_2O_2$- and cryo-induced apoptosis in mouse and bovine embryos. Our findings suggest that apoptosis in mouse and bovine embryos might be controlled by modulation of $K^+$ channels which are highly expressed in a given cell type.

Effects of Subatmospheric Pressure Storage on Ultrastructural Changes in the Chloroplasts of Apple Fruit (사과과실 엽록체의 초미세구조 변화에 미치는 감압저장의 영향)

  • Ha, Young-Sun;Sohn, Tae-Hwa
    • Applied Biological Chemistry
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    • v.28 no.3
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    • pp.174-181
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    • 1985
  • Ultrastructural changes in the chloroplast of apple fruits was examined with Malus Pumila Mill var. Fugi harvested during preclimacteric stage the subatmospheric pressure storage condition at 380mmHg, $20^{\circ}C$. Electron micrographs obtained from the fruits stored at 760mmHg, $20^{\circ}C$ indicated that the initiation of ultrastructural changes is in parallel with the onset of respiratory climacteric. Chloroplast of the apple fruits stored for 25 days is vacuolated and the lamellar system are a little disorganized. Chloroplast of the apple fruits stored for 50 days is showing extensive vacuolation, the lamellar system are completely disorganized and the plasma membrane has pulled away from the cell wall. In the case of subatmospheric presssure storage caused to delay the onset of ultrastructural changes in chloroplast such as vacuolation, disorganization of lamellar system and dispersal of stroma lamella about 2 months. These results suggested that ripening of apple fruits is correlated not only to the activity of respiration but to the involvement of ultrastructural changes in fruit cells. The evidences obtained here led to the conclusion that subatmospheric pressure treatment extends storage life of apple fruits by inhibiting ultrastructural changes of cell organelles in concomitance with respiratory activity.

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Storage-life and Palatability Extension of Betula platyphylla Sap Using Lactic Acid Bacteria Fermentation (유산균 발효를 이용한 자작나무 수액의 저장성 및 기호성 증진 기술)

  • Kim, Jong-Ho;Lee, Woon-Jong;Cho, Youn-Won;Kim, Kwang-Yup
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.38 no.6
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    • pp.787-794
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    • 2009
  • In this study, a new method for extending storage-life and palatability of Betula platyphylla sap by applying lactic acid bacteria fermentation was developed. The fluids of saps were filtered through 0.22 ${\mu}m$ membrane filter and each fermented by 8 different lactic acid bacteria which are Lactobacillus acidophilus, Lactobacillus brevis, Leuconostoc mesenteroides, Leuconostoc lactis, Lactococcus lactis, Pediocossus pentosaceus, Pediococcus dextrinicus, Streptococcus thermophilus. All the tested lactic acid bacteria except P. dextrinicus grew fast up to $10^6{\sim}10^7cfu/mL$ levels and lowered pH down to about pH 4 levels in 48 hours in both saps. The produced organic acids and lowered pH level inhibited the growth of spoilage microorganisms almost completely for 2 weeks during storage at room temperature. Addition of xylitol in the saps before fermentation accelerated the growth of lactic acid bacteria and increased the sweetness and overall taste of final product. The filtration process did not affect the mineral compositions of Betula platyphylla saps. Also the compositions and amounts of minerals showed very minor differences before and after fermentation in Betula platyphylla saps inoculated with L. acidophilus. By applying lactic acid fermentation to extend storage-life of tree saps instead of heat treatment, it was possible to keep natural minerals in active forms without any modifications.

Studies on the Antitumor Activity of Gamisoam-san via Suppressing Angiogenesis and Growth Factor Expression (혈관신생 및 이식암세포증식 억제를 통한 가미소암산의 항암작용연구)

  • Yoon Sung Chan;Ahn Seong Hun;Mun Yean Ja;Kim Jin Kyeong;Choo Young Kug;Jung Kyu Yong;Kim Yeong Mok;Woo Won Hong
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.17 no.4
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    • pp.969-979
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    • 2003
  • Gamisoamsan is a prescription originated in Soamsan which is known as an anti-cancer remedy in the traditional Korean Medicine. To enhance the synergic effects of anti-cancer activity of Soamsan, this study reconstituted the original components of Soamsan with a slight modification and produced a novel herbal remedy, namely Gamisoamsan. To investigate the effects of Gamisoamsan on anti-cancer reaction, I studied the effects of Gamisoamsan on angiogenesis via chorioallantoic membrane (CAM) assay, corneal neovascularization assay and the effects on expression of growth factor which are VEGF, TGF-β, bFGF and IMUP-1. Anti-cancer effects of Gamisoamsan was also abserved through hematological parameters, tumor volume and survival rate in mice. Gamisoamsan inhibited embryonic angiogenesis of blood vessels in CAM assay and inhibited neovascularization of ral cornea. Gamisoamsan reduced cell proliferation in HT1080 cells and IC50 was 2.18 ㎎/㎖ Gamisoamsan reduced the expression of VEGF, TGF-β, bFGF and IMUP-1 which was known as vascular growth factor and this effects of Gamisoamsan was predominant than VP-16. The treatment of Gamisoamsan decreased the CT-26 cell inoculated-tumor volume in mice colon adenocarcinoma and increased mice survival which was inoculated CT-26 cells. The results of the present study suggest that Gamisoamsan extracts has a potential anti-tumor activity and may be an useful remedy to prevent and/or treat cancer.

A Case of Membranoproliferative Glomerulonephritis Type II(Dense-Deposit Disease) (막증식성 사구체신염 제 II 형(Dense-Deposit Disease) 1례)

  • Lee Suk-Jin;Moon Jae-Hoon;Kang Mi-Seon;Song Min-Seob;Chung Woo-Yeong
    • Childhood Kidney Diseases
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    • v.7 no.2
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    • pp.204-210
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    • 2003
  • Membranoproliferative glomerulonephritis type II(MPGN II), also called dense deposit disease, was first described by Berger and Galle in 1963. The diagnosis of MPGN II is based on electron-microscopic finding of an intensely electron-dense substance which replaces the lamina densa of the glomerular basement membrane. Although the etiology and pathogenesis of MPGN II are unknown, it frequently progresses to end-stage renal failure. Typically in MPGN II, hypocomplementemia due to activation of the alternative complement pathway is present. In addition, the association of MPGN II with partial lipodystrophy and complement abnormalities is well documented. The relationship between these associated features and the patient's renal functional outcome is not clear. With respect to the therapy for MPGN II, an alternate-day prednisolone regimen was shown to be effective. Various treatment modalities, including immunosuppression with corticosteroids, cytotoxic drugs and cyclosporin A, anticoagulants and antiplatelet therapies are used, either alone or in combination, with varying degrees of success. The purpose of this paper is to present a case of MPGN II from a 7 years old girl with paroxysmal supraventricular tachycardia(PSVT).

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Water Extract of Allium sativum L. Induces Apoptosis in Human Leukemia U937 Cells through Reactive Oxygen Species Generation (마늘 열수 추출물의 활성산소종 생성을 통한 인체백혈병세포의 apoptosis 유발)

  • Choi, Woo-Young;Chung, Kyung-Tae;Yoon, Tae-Kyung;Choi, Byung-Tae;Lee, Yong-Tae;Lee, Won-Ho;Ryu, Chung-Ho;Choi, Yung-Hyun
    • Journal of Life Science
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    • v.17 no.12
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    • pp.1709-1716
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    • 2007
  • The health benefits of garlic (Allium sativum L.) are derived from a wide variety of components and from the different ways it is administered. The known health benefits of garlic include cardiovascular protective effects, stimulation of immune function, reduction of blood glucose level, protection against microbial, viral and fungal infections, as well as anticancer effects. In the present study, it was examined the effects of water extract of A. sativum (WEAS) on the growth of cultured human tumor cells in order to investigate its anti-proliferative mechanism. Treatment of WEAS to tumor cells resulted in the growth inhibition, especially in leukemia cells, which was associated with induction of G2/M arrest of the cell cycle and apoptosis. In order to further explore the critical events leading to apoptosis in WEAS-treated U937 human leukemia cells, the following effects of WEAS on components of the mitochondrial apoptotic pathway were examined: generation of reactive oxygen species (ROS), alteration of the mitochondrial membrane potential (MMP), and the expression changes of Bcl-2 and IAP family proteins. The cytotoxic effect of WEAS was mediated by its induction of apoptosis as characterized by the occurrence of DNA ladders, apoptotic bodies and chromosome condensation in U937 cells. The WEAS-induced apoptosis in U937 cells was correlated with the generation of intracellular ROS, collapse of MMP, activation of caspase-3 and down-regulation of anti-apoptotic proteins. The quenching of ROS generation with antioxidant N-acetyl-L-cysteine conferred significant protection against WEAS-elicited ROS generation, caspase-3 activation, G2/M arrest and apoptosis. In conclusion, the present study reveals that the cellular ROS generation plays a pivotal role in the initiation of WEAS-triggered apoptotic death in U937 cells.

Role of FAK Phosphorylation in Cobalt Chloride-Induced Epithelial-to-Mesenchymal-Like Transition (Cobalt chloride에 의해 유도되는 상피-중간엽 이행에서의 국소부착 단백질의 인산화의 역할 규명)

  • Nam, Ju-Ock
    • Journal of Life Science
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    • v.21 no.2
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    • pp.286-291
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    • 2011
  • Hypoxia is a common condition found in a wide range of solid tumors and is often associated with metastasis and poor clinical outcomes. In the present study, we found that HIF-$1{\alpha}$ was induced by cobalt chloride (500 ${\mu}M$) treatment on human lung cancer cells, A549 and H460, for 24 hr. However, cobalt chloride (500 ${\mu}M$) did not affect cell proliferation of A549 and H460 in 48 hr. Cobalt chloride (500 ${\mu}M$) additionally induced epithelial-to-mesenchymal-like transition (EMT) such as reduced E-cadherin expression and increased ${\alpha}$-SMA expression. These results were confirmed by immunofluorecence experiment in H460 cells. E-cadherin was localized on the outer cell membrane. However, when the cells were treated with 500 ${\mu}M$ cobalt chloride for 24 hr, diffuse E-cadherin staining was observed, characteristic of a migratory mesenchymal phenotype. We also found that cobalt chloride induced integrin ${\beta}3$ expression and FAK phosphorylation in human lung cancer cells using western blotting and FACS anlaysis. Our data suggest that integrin ${\beta}3$-induced FAK phosphorylation may be developed into target molecules for blocking tumor metastasis.

The Effects of Fucoidan on the Activation of Macrophage and Anticancer in Gastric Cancer Cell (Fucoidan의 면역세포 활성 및 위암 세포주에서의 항암효과)

  • An, In-Jung;Cho, Sung-Dae;Kwon, Jung-Ki;Kim, Hye-Ri;Yu, Hyun-Ju;Jung, Ji-Youn
    • Journal of Food Hygiene and Safety
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    • v.27 no.4
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    • pp.406-414
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    • 2012
  • This study was designed to investigate the effect of fucoidan on the activation of macrophage and on induction of apoptosis in AGS cell. To measure the activity of macrophages, NO and TNF-${\alpha}$ assays were performed in Raw 264.7 cell. Treatment with fucoidan significantly increased production of NO and TNF-${\alpha}$, indicating activation of macrophages. The result of MTT assay shows that cell viability was significantly decreased in a dose and time-dependent manner. Fucoidan increased to enhance mitochondrial membrane permeability, as well as the cytochrome c release from the mitochondria. Fucoidan decreased Bcl-2 and XIAP expression, whereas the expression of Bax was increased in a time-dependent manner compared to the control. In addition, the active forms of caspase-9 were increased, and the inactivation of Akt was decreased in a time-dependent manner. Caspase inhibitor, z-VAD-FMK, canceled the apoptosis of fucoidan, expression of Bax and caspase-9 were decrease. These results indicate that fucoidan induces activation of macrophage and apoptosis through activation of caspase on AGS cell.

Case of Anterior Segment Ischemia after Two Vertical Rectus Muscles Surgery (두 개의 수직근수술 후 발생한 전안부허혈 1예)

  • Ahn, Jayoung;Kim, Gyu Nam;Kim, Seong Jae;Chung, In Young;Seo, Seong Wook;Yoo, Ji Myong
    • Journal of The Korean Ophthalmological Society
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    • v.59 no.12
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    • pp.1195-1200
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    • 2018
  • Purpose: To report a case of anterior segment ischemia after superior and inferior rectus muscle surgery in a patient with superior rectus muscle enlargement. This is the first report in the Republic of Korea of anterior segment ischemia after two rectus muscles surgery. Case summary: An 80-year-old male was referred to our clinic with a 3 years history of diplopia. The patient had 30 prism diopters right eye hypertropia in the primary position and downgaze limitation. The orbital computed tomography scan revealed enlargement of the right superior rectus muscle. He underwent right superior rectus recession and inferior rectus resection. On postoperative day 2, slit lamp examination revealed diffuse corneal edema, Descemet's membrane folding, an aqueous flare and a dilated pupil. Treatment with 45 mg oral steroid was initiated, and 1% prednisolone acetate and 0.5% Levofloxacin eye drops were administered. At postoperative 1 month, the patient was orthophoric in the primary position, and there was no corneal edema, pupil abnormality or aqueous flare. Conclusions: Base on the present case, the possibility of anterior segment ischemia should be considered after even two muscles surgery, and older patients with vertical muscle surgery should be considered more carefully.