• Title/Summary/Keyword: calcium concentration

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Studies on the $\beta$-Galactosidase from Thermphilic Bacterium - Physiological Characteristics of the Selected Thermophile - (고온성 세균의 $\beta$-Galactosidase에 관한 연구 ( I ) - 분리고온균의 생리적 특성 -)

  • 이종수;오만진;이석건;김찬조
    • Microbiology and Biotechnology Letters
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    • v.11 no.1
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    • pp.5-13
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    • 1983
  • This experiment was carried out to elucidate the thermotolerant properties of a thermophilic bacterium which isolated from soils of the hot springs area and selected for the $\beta$-galactosidase production. Biochemical and physiological characteristics of this strain were studied, including the investigation on the fatty acid composition for its neutral fats. The results obtained were summarized as follows. 1. This bacterium was identified as a strain belong to the genus Thermus. 2. Optimal temperature and pH for growth of this strain were $65^{\circ}C$ and pH 6.5 respectively, and it was found to be an absolute thermophilic bacterium which could not grow at 37$^{\circ}C$. 3. No growth was obtained in the medium which contained more than 1.0% of sodium chloride. 4. The tolerable concentration of antobiotics were 10$\mu\textrm{g}$ of penicillin G per $m\ell$ of medium and 0.5$\mu\textrm{g}$ of chloramphenicol per $m\ell$ respectively 5. This strain had autotrophilic requirements for calcium-pantothenate and pyridoxine-HCO as an-essential factor and for niacin as a stimulative factor. 6. Fatty acid composition of neutral fats of the strain was palmitic acid. 60.20%; lauric acid, 11.8%; myristic acid, 7.56%, behenic acid, 4.25%; capric acid, 1.77%; stearic acid, 2.13%; arachidic acid, 1.53%; and others unidentified, 10.7%.

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Human Neutrophil Elastase: Rapid Purification, Metal binding Stoichiometry and Modulation of the Activity by Chelating Agents (사람의 백혈구 내에 있는 Elestase: 순수부리, 금속이온의 화학량, 그리고 Chelating 효과에 의한 활성도 조절)

  • Kang, Koo-Il
    • The Korean Journal of Pharmacology
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    • v.24 no.1
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    • pp.111-123
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    • 1988
  • Neutrophil elastases were purified by a three step procedure consiting of one Sephadex G-75 and two HPLC elutions. The elastases cross-reacted with antibodies to human neutrophil elastase. Three bands with molecular weights between 26,000 and 29,700 were observed by gel electrophoresis. At each stage of purification the quantity of Zn increased, reaching molar ratio of 2:1 with elastase in the most purified samples. Calcium content. was seletively elevated during the earlier stages of purification but decreased to a ratio of 0.25 to 1 with elastase at the final step of purfication. Neutrophil elastase could be inhibited by EDTA, EGTA and 1,10-phenanthroline. EGTA inhbition was noncompetitive inhibition and reversible only if the time of preincubation was relatively short, indicating the instability of the apoenzyme. The concentration of chelator required to show significant inhibition of elastase was also dependent upon the stage of purity and the ionic strength of the reaction mixture. Inhibition by EGTA, followed by the removal of EGTA, could be reversed by Zn. In the presence of EGTA the enzyme could be returened to full activity by the addition of Zn, Mn and Ca, but not Mg or Na. All of the above evidence strongly supports human neturophil elastase could be a metalloenzyme as well as a serine protease.

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Cooperation of $G{\beta}$ and $G_{\alpha}q$ Protein in Contractile Response of Cat Lower Esophageal Sphincter (LES)

  • Sohn, Uy-Dong;Lee, Tai-Sang
    • The Korean Journal of Physiology and Pharmacology
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    • v.7 no.6
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    • pp.349-355
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    • 2003
  • We previously shown that LES contraction depends on $M_3$ receptors linked to PTX insensitive $G_q$ protein and activation of PLC. This results in production of $IP_3$, which mediates calcium release, and contraction through a CaM dependent pathway. In the esophagus ACh activates $M_2$ receptors linked to PTX sensitive $G_{i3}$ protein, resulting in activation of PLD, presumably, production of DAG. We investigated the role of PLC isozymes which can be activated by $G_q$ or $G{\beta}$ protein on ACh-induced contraction in LES and esophagus. Immunoblot analysis showed the presence of 3 types of PLC isozymes, $PLC-{\beta}1$, $PLC-{\beta}3$, and $PLC-{\gamma}1$, but not $PLC-{\beta}2$, $PLC-{\beta}4$, $PLC-{\gamma}2$, $PLC-{\delta}1$, and $PLC-{\delta}2$ from both LES and esophageal muscle. ACh produced contraction in a dose dependent manner in LES and esophageal muscle cells obtained by enzymatic digestion with collagenase. $PLC-{\beta}1$ or $PLC-{\beta}3$ antibody incubation reduced contraction in response to ACh in LES but not in esophageal permeabilized cells, but $PLC-{\gamma}1$ antibody incubation did not have an inhibitory effect. The inhibition by $PLC-{\beta}1$ or $PLC-{\beta}3$ antibody on Ach-induced contraction was antibody concentration dependent. The combination with $PLC-{\beta}_1$ and $PLC-{\beta}_3$ antibody completely abolished the contraction, suggesting that $PLC-{\beta}1$ and $PLC-{\beta}3$ have a synergism to inhibit the contraction in LES. $PLC-{\beta}1$, -${\beta}3$ or -${\gamma}1$ antibody did not reduce the contraction of LES cells in response to DAG ($10^{-6}$ M), suggesting that this isozyme of PLC may not activate PKC. When $G_{q/11}$ antibody was incubated, the inhibitory effect of the incubation of PLC ${\beta}3$, but not of PLC ${\beta}_1$ was additive (Fig. 6). In contrast, when $G_{\beta}$ antibody was incubated, the inhibitory effect of the incubation of PLC ${\beta}_1$, but not of PLC ${\beta}_3$ was additive. This data suggest that $G_{q/11}$/11 or $G{\beta}$ may activate cooperatively different PLC isozyme, $PLC{\beta}_1$ or $PLC{\beta}_3$ respectively.

The Efficiency of Vascular Embolization Using Alginate Gel : An Experimental Study in Rabbit (알지네이트 젤을 이용한 혈관 색전술의 유용성 : 토끼에서의 실험적 연구)

  • Lee, Woo-Baek;Kang, Yeong-Han;Kim, Jong-Ki
    • Journal of radiological science and technology
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    • v.32 no.1
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    • pp.61-67
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    • 2009
  • Purpose : The purpose of this study was to investigate the applicability of poly-L-guluronic alginate (PGA) gel in vascular embolization with angiography simulation. Materials and Methods : To prepare a gel-forming PGA from no guluronate-rich Laminaria japonica, a new acid hydrolysis method was employed with a lower HCL concentration (0.03 M) and a shorter treatment time (5 min). The obtained PGAs were selected based on gel stability and viscosity. Glass aneurysm model was used to simulate gel embolization in vitro. Then, finally, the PGA was used to embolize the renal vascular system by using a rabbit model and angiography. Results : Glass aneurysm model was made to simulate gel embolization procedure. PGA solution was injected from pump through 2-way catheter. Subsequent injection of $CaCl_2$ successfully formed gels inside aneurysm model that conforming to its inner contour. In rabbit model, first, renal artery and aorta leading to the right kidney were ligated to block blood flow, then conventional contrast agent was injected through aorta to check the arterial patency to the left kidney. In sequential artery injection method, PGA and $CaCl_2$ were injected through renal artery sequentially via a single catheter. Re-injection of contrast agent after removing ligated aorta showed blood flow to the right kidney but no flow in the left kidney. This result demonstrated a complete blocking of blood flow due to gel formation in vascular bed of the left kidney. Conclusion : Instillation of calcium alginate into aneurysm model and arterial system in vivo produced an embolization that better fills and conforms to the contour of aneurysms or blocking vascular bed completely. Therefore, PGA was effective endovascular occlusion materials and provide an efficiency of vascular angiography.

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Changes of enzyme activity in the hemolymph and hepatopancreas of the abalone, Haliotis discus hannai (Ino, 1953) exposed to cadmium (카드뮴 노출에 따른 북방전복, Haliotis discus hannai (Ino, 1953) 의 hemolymph 및 hepatopancreas의 효소활성의 변화)

  • Min, Eun-Young;Lee, Jung Sick;Kwak, Ihn-Sil;Kim, Jae Won;Kang, Ju-Chan
    • The Korean Journal of Malacology
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    • v.30 no.1
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    • pp.41-49
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    • 2014
  • This study was conducted to investigate the effects of cadmium (Cd) exposure on biochemical factors in the hemolymph and hepatopancreas of the abalone, Haliotis discus hannai. The abalone were exposed to 0, 5, 10, 20 and 40 ${\mu}g/L$ Cd for 4 weeks. The phenoloxidase (PO) activity was decreased in hemolymph of abalone exposed to 40 Cd ${\mu}g/L$ for 4 weeks compared to the control (P < 0.05). The hemolymph enzymes, alkaline phosphatase (ALP), aspartate aminotransferase (AST) and alanine aminotransferase (ALT) activities were markedly elevated in 40 Cd ${\mu}g/L$ after 4 weeks. The hemolymph calcium concentrations were significantly decreased in 20 and 40 Cd ${\mu}g/L$ for 4 weeks. Hepatopancreas superoxide dismutase (SOD) and catalase (CAT) activities were significantly increased by Cd. SOD was increased in both 20 and 40 Cd ${\mu}g/L$ and CAT, in 40 Cd ${\mu}g/L$ after 2 weeks (P < 0.05). These results suggested that the abalone SOD and CAT including PO may serve as a protective mechanism against oxidative stress by Cd. We conclude that a Cd concentration, 40 ${\mu}g/L$ in water may curtail hemolymph homeostasis and anti-oxidative reactions in abalone hepatopancreas. From these results, these biochemical factors may represent a convenient method of monitoring heavy metal pollution in coastal areas.

Effects of 2-deoxy-D-glucose and quercetin on osteoblastic differentiation and mineralization in irradiated MC3T3-E1 cells (2-deoxy-D-glucose와 quercetin이 방사선조사 MC3T3-E1 골모세포주의 분화와 석회화에 미치는 영향)

  • Ahn, Hyoun-Suk;Kim, Kyoung-A;Koh, Kwang-Joon
    • Imaging Science in Dentistry
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    • v.36 no.4
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    • pp.189-198
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    • 2006
  • Purpose: To investigate the in vitro response of MC3T3-E1 osteoblastic cells to X-ray in the presence and absence of 2 deoxy-D-glucose (2-DG) and quercetin (QCT). Materials and Methods: The MC3T3-E1 cells were cultured in an ${\alpha}-MEM$ supplemented with 5 mM 2-DG or $10{\mu}M$ QCT and then the cells were incubated for 12 h prior to irradiation with 2, 4, 6, and 8Gy using a linear accelerator (Mevaprimus, Germany) delivered at a rate of 1.5 Gy/min. At various times after the irradiation, the cells were processed for the analyses of proliferation, viability, cytotoxicity, and mineralization. Results: Exposure of the cells to X-ray inhibited the tritium incorporation, 3-(4, 5-dimethylthiazol-2yl-)-2, 5-diphenyl tetrazolium bromide (MTT)-reducing activity, and alkaline phosphatase (ALP) activity, and caused cytotoxicity and apoptosis in a dose-dependent manner of the X-ray. This effect was further apparent on day 3 and 7 after the irradiation. RA+2-DG showed the decrease of DNA content, cell viability, and increase of cytotoxicity rather than RA. ALP activity increased on day 7 and subsequently its activity dropped to a lower level. 2-DG suppressed the calcium concentration, but visual difference of number of calcified nodules between RA and RA+2-DG was not noticed. RA+QCT showed the increase of DNA content, cell viability, but decrease of cytotoxicity and subG1 stage cells in the cell cycle, and increased calcified nodules in von Kossa staining rather than the RA. ALP activity showed significant increases on day 7 and subsequently its activity dropped to a lower level. Conclusion: The results showed that the 2-DG acted as a radiosensitizing agent and QCT acted as a radiosensitizing agent respectively in the irradiated MC3T3-E1 osteoblast-like cells.

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Effect of Extracellular Potassium on Delayed Rectifier Potassium Channel Proteins of KCNQ3 and KCNQ5 in Familial Hypokalemic Periodic Paralysis (가족성 저칼륨성 주기성 마비에서 세포외 칼륨농도가 지연성 정류형 채널을 형성하는 KCNQ3와 KCNQ5 단백질에 미치는 효과)

  • Kim, Sung-Jo;Kim, Dong-Hyun;Kim, June-Bum
    • Journal of Life Science
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    • v.19 no.10
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    • pp.1484-1488
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    • 2009
  • Familial hypokalemic periodic paralysis (HOKPP) is an autosomal dominant muscle disorder characterized by episodic attacks of muscle weakness with concomitant hypokalemia. Mutations in either a calcium channel gene (CACNA1S) or a sodium channel gene (SCN4A) have been shown to be responsible for this disease. The combination of sarcolemmal depolarization and hypokalemia has been attributed to abnormalities of the potassium conductance governing the resting membrane potential. To understand the pathophysiology of this disorder, we examined both mRNA and protein levels of delayed rectifier potassium channel genes, KCNQ3 and KCNQ5, in skeletal muscle fibers biopsied from patients with HOKOur results showed an increase in the cytoplasmic level of KCNQ3 protein in patients' cells exposed to 50 mM external concentration of potassium. However, mRNA levels of both channel genes did not show significant change in the same condition. Our results suggest that long term exposure of skeletal muscle cells in HOKPP patients to high extracellular potassium alters the KCNQ3 localization, which could possibly hinder the normal function of this channel protein. These findings may provide an important clue to understanding the molecular mechanism of familial hypokalemic periodic paralysis.

Combined effects of copper and temperature on Hematological constituents in the Rock fish, Sebastes schlegeli (조피볼락, Sebastes schlegeli의 혈액학적 성분변화에 미치는 구리 및 온도의 복합적 영향)

  • Baeck, SuKyong;Min, EunYoung;Kang, Ju-Chan
    • Journal of fish pathology
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    • v.27 no.1
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    • pp.57-65
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    • 2014
  • Copper ($CuSO_4$) has been widely used to control algae and pathogens in fish culture ponds. However, its toxic effects on fish depend not only on its concentration in water but also on water quality. The susceptibility of the rockfish, Sebastes schlegeli to copper was evaluated at three water-temperatures (WT; 18, 23 and $28^{\circ}C$) for 4 days. After the exposure of two copper concentrations (100 and $200{\mu}g/L$), a hematological effect was exerted on rockfish, by causing changes in red blood cell count and hematocrit value at $28^{\circ}C$. Total protein levels of the fish showed a tendency of co-increase with glucose depend on the WT, after copper exposure. However, the plasma calcium and magnesium levels were significantly increased at $200{\mu}g/L$ copper, regardless of the WT. Enzymes activities including ALT and LDH in serum were also significantly increased depend upon the copper treatment only. This indicates that inorganic components and enzymes activities were sensitive indexes to stress by toxicant such as copper. The cortisol levels were significantly elevated by both WT rising and copper treatment in serum of rock fish. In conclusion, these changes can be seen as an initial response to temperature stress and as a sustaining response to copper exposure. The present findings suggest that a simultaneous stress by temperature change and copper exposure could accelerate an alteration of hematological and plasma biological parameters in the rockfish.

Isolation and Characterization of a β-galactosidase Producing Thermophilic Bacterium (β-galactosidase를 생성하는 고온성(高溫性) 세균(細菌)의 생리적(生理的) 특성(特性)에 관한 연구(硏究))

  • Lee, Jong Soo;Kim, Chan Jo
    • Korean Journal of Agricultural Science
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    • v.9 no.1
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    • pp.377-386
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    • 1982
  • This experiment was carried out to elucidate the thermotolerant properties of a thermophilic bacterium which was isolated from soils of the hot springs area and selected for the ${\beta}$-galactosidase production. This bacterium was identified as a strain belong to the genus Thermus. Biochemical and physiological characteristics of this strain were studied, including the investigation of the fatty acid composition of its neutral fats. The results obtained were summarized as follows. 1. Optimal temperature and pH for growth of this strain were $65^{\circ}C$ and pH 6.5 respectively, and it was found to be an absolute thermophilic bacterium which could not grow at the temperature below $43^{\circ}C$. 2. No growth was obtained in the medium which contained more than 1.0% of sodium chloride. 3. The tolerable concentration of antibiotics were 10mg of penicillin G per ml of medium and 0.5mg of chloramphenicol per ml respectively. 4. This strain had auxotrophilic requirements for calcium-pantothenate and pyridoxin-HCl as an essential factor and for niacin as a stimulative factor. 5. Yellow pigment was released into the liquid culture of this strain, which showed maximum absorption at 420 nm. 6. Fatty acid composition of neutral fats of the strain was palmitic acid, 60.20%; lauric acid, 11.80%; myristic acid, 7.56%; behenic acid, 4.25%; capric acid, 1.77%; stearic acid, 2.13%; arachidic acid, 1.53%; and others unidentified, 10.7%.

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Evaluation of Salt Tolerance of Liriope platyphylla and Pachysandra terminalis to Deicing Salt (CaCl2) Concentration in Winter (겨울철 제설제(CaCl2)농도처리에 따른 맥문동과 수호초의 내염성 평가)

  • Ju, Jin-Hee;Hui, Xu;Park, Ji-Yeon;Choi, Eun-Young;Yoon, Yong-Han
    • Korean Journal of Environment and Ecology
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    • v.30 no.4
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    • pp.651-657
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    • 2016
  • It is important to know know deleterious impact of deicing salt on plants for guidelines of planting along roads. The aim of this study was to determine the effect of calcium chloride ($CaCl_2$) on the growth and physiological characteristics of Liriope platyphylla and Pachysandra terminalis. The plants were grown from November of 2015 to March of 2016 in pots containing growing media with $CaCl_2$ at 0% (Control), 0.5%, 1.0%, 3.0%, and 5.0% (based on the weight). While plant growth and photosynthetic activity were significantly decreased in both plant species grown on the media with $CaCl_2$, the degree of sensitivity to $CaCl_2$ differed. The plant growth of Liriope platyphylla was considerably injured under higher than 3.0% of $CaCl_2$, whereas Pachysandra terminalis was all dead under higher than 1.0% of $CaCl_2$. This results indicate that Liriope platyphylla has higher degree of tolerance to the deicing salt than Pachysandra terminalis.