• Title/Summary/Keyword: Alkaline phosphatase

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Cloning of the Alkaline Phosphatase Gene from Kluyveromyces fragilis

  • Kim, Jong-Guk;Hwang, Seon-Kap;Kwon, Kaeg-Kyu;Nam, Joo-Hyun;Hong, Soon-Duck;Seu, Jung-Hwn
    • Journal of Microbiology and Biotechnology
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    • v.2 no.4
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    • pp.237-242
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    • 1992
  • In order to clone the gene coding for alkaline phosphatase in the yeast Kluyveromyces fragilis, a genomic library was constructed using the yeast-E. coli shuttle vector pHN114 as a cloning vector. From the genomic library, a clone carrying the gene was isolated and the plasmid was designated as pSKH101. A restriction enzyme map was made using this plasmid. Subcloning experiments and complementation studies showed that alkaline phosphatase was active only in the original 3.1 kb insert. Southern hybridization analysis confirmed that the cloned DNA fragment was derived from K. fragilis genomic DNA. Using a minicell experiment, the product of the cloned gene was identified as a protein with a molecular weight of 63 KDa. A 0.6 kb HindIII fragment, which showed promoter activity, was isolated using the E. coli promoter-probe vector pKO-1.

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Influences of $\alpha$-Tocopherol on the Toxicity of Vitamin A Acetate in Rats (랏트에 있어서 Vitamin A Acetate의 독성에 미치는 $\alpha$-Tocopherol의 영향)

  • 안영근;김성오;오연준;박영길
    • Environmental Analysis Health and Toxicology
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    • v.4 no.3_4
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    • pp.77-86
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    • 1989
  • Influences of u-tocopherol on the toxicity of vitamin A acetate in male rats were studied. The obtained results are as follows; 1) The administration of vitamin A acetate 500,000 IU/Kg i.p. twice at 3 days interval decreased the liver weight/body weight and increased the spleen weight/body weight, and increased activities of SGOT and alkaline phosphatase, and also increased BUN and creatinine. 2) ${\alpha}$-Tocopherol administered together with vitamin A acetate as given as the above 1) poteniated the increase of SGOT activity caused by vitamin A acetate and reduced the increase of alkaline phosphatase activity and creatinine which were caused by vitamin A acetate. 3) The administration of vitamin A acetate 500,000 IU/Kg i.p. twice a week for 4 weeks showed remarkable decrease of body weight gain and the effect of it was larger in later stage than in early. It increased significantly liver weight/body weight and further increased the activities of SGOT, SGPT and alkaline phosphatase, and showed no influnence on BUN and creatinine. 4) ${\alpha}$-Tocopherol administered together with vitamin A acetate as given as the above 3) reduced the decrease of body weight gain caused by vitamin A acetate, and potentiated remarkably the increased activities of SGOT and alkaline phosphatase which were caused by vitamin A acetate.

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BIOCHEMICAL CHARACTERISTICS OF HUMAN PERIODONTAL LIGAMENT CELLS IN VITRO (치주인대세포(齒周靭帶細胞)의 생화학적(生化學的) 특이성(特異性)에 대(對)한 연구(硏究))

  • Cho, Soung-Wook;Cha, Kyung-Suk
    • The korean journal of orthodontics
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    • v.22 no.1
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    • pp.273-283
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    • 1992
  • To find out the differences between periodontal ligament cells (PDL cells) and gingival fibroblast cells (GFB cells), alkaline phosphatase, a marker enzyme for osteoblast, was used to measure the activities and $^{45}CaCl_2$ isotope was used to find out cellular and release of $^{45}Ca$, a requisite for bone formation,. PDL cells and GFB cells from 1 to 5 passages were also measured in alkaline phosphatase activity assay. By the use of above methods, followings were concluded that the PDL cells and the GFB cells have characteristics that are different from each other. In that PDL cells showed large amount of calcium uptake and large amount of calcium release in initial stage, they seem to possess characteristics which are similar to osteoblast-like cells. 1. The PDL cells, in contrast to the gingival fibroblast, showed exceedingly high alkaline phosphatase activity which was highest at the second passage, decreasing thereon. But gingival fibroblasts cells showed no distinct differences in alkaline phosphatase activity as the passage were elapsed. 2. For both PDL cells and GF cells, the $^{45}Ca$ uptake was greatest at 2 hours period. The PDL cells showed higher measuring than GFB cells through out the whole time period. 3. Whereas the GFB cells showed slow increase of $^{45}Ca$ release as time relapsed, the PDL cells showed rapid increase of $^{45}Ca$ release.

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Properties of a Thermolabile Alkaline Phosphatase from the Marine Bacterium Vibrio sp. M-96 (해수에서 분리한 Vibrio sp. M-96 균주의 열감수성 alkaline phosphatase 성질)

  • Park, Moon-Kyung;Jin, Deuk-Hee;Kim, Joong-Kyun;Kong, In-Soo;Kim, Kwang-Hyeon;Hong, Yong-Ki
    • Journal of Life Science
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    • v.6 no.3
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    • pp.198-203
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    • 1996
  • A thermolabile alkaline phosphatase has been purified through steps of osmotic shock, ammonium sulfate salting-out, and DAEA-cellulose chromatography from the cultured broth of the marine Vibrio sp. M-96 strain. The optimal temperature for the enzyme activity was 35$\circ$C. The optimal pH was pH11.0, and the range of pHstability was pH10.4 to 12.0. Thermal inactivation occured within 6 mintes at 60$\circ$C. The enzyme was considerably inactivated by 0.1mM concentrations of Hg$^{2+}$, Ni$^{2+}$ and Zn$^{2+}$, whereas activated up to 234% by 1mM of Mn$^{2+}$. The activation energy and deactivation energy by the Arrhenius equation were 4.02 Kcal/mol and 9.098 Kcal/mol, respectively. The Km and Vmax values of the enzyme for p-introphenylphosphate were found to be 0.0465mM and 0.001334mM/min, respectively. Active form of the enzyme had a molecular weight of 57,000 dalton determined by the Sephadex G-200 gel filtration method.

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On the Activity of Phosphatase in the Endometrium of the Rat Uterus During Early Pregnancy (초기 임신 기간중 흰쥐 자궁 내막조직의 Phosphatase 활성에 관하여)

  • Kim, Sung-Rye;Cho, Wan-Kyoo
    • Clinical and Experimental Reproductive Medicine
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    • v.8 no.2
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    • pp.1-11
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    • 1981
  • The quantitative analyses of the phosphatase activity in the endometrium of the rat ovariectomized on Day 2 of pregnancy was carried out in comparison with the intact one, in order to investigate the hormonal dependency of the uterus prior to the implantation, and to study the phosphatase activity in the endometrial tissues in vitro incubated in different acidity of the medium. The results obtained were as follows: 1. The activity of the total phosphatase was the highest at Day 3 of pregnancy of the intact animals irrespective of acidity of the medium. However, the ovariectomized rat showed its peak somewhat delayed. The time of the highest activity of the enzymes was matched with the time of high secretion of the ovarian hormones. 2. The activity of acid phosphatase in the endometrium was twice or four times as much high as that of neutral or alkaline phosphatase, respectively. 3. The activity of alkaline phosphatase was rather steady in Day 3 through Day 5 of the pregnancy of the rat intact or ovariectomized but with low level compared to those of other phosphatase. 4. The present re~lt indicated more important role by $Mg^{2+}$-dependent phosphatase than by $K^+$-dependent one for the preparation for decidualization.

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Vertical Profiles of Alkaline Phosphatase Activity in Dam Reservoirs and its Relation with Microbial Parameters (댐 저수지에서 alkaline phosphatase 활성의 수직변화와 미생물 요인들과의 상관관계)

  • Nakagawa, Ayumi;Kagawa, Hisanori;Hiroshi, Hirotani
    • Korean Journal of Ecology and Environment
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    • v.37 no.4 s.109
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    • pp.406-410
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    • 2004
  • The alkaline phosphatase activity (APA) of two dam reservoirs and inflowing streams were measured monthly in 2000. During summer months in 2001, the vertical profiles of APA and related parameters were also examined in one of the reservoirs. The APA was relatively high during the summer season in the epilimnion while it was almost invariable in the hypolimnion. A small increase in APA was observed at just above the bottom. The APA fluctuation was independent of the concentration of soluble reactive phosphorus. It was assumed that APA is not indicative of the phosphorus availability status. An examination of size-fractionated samples suggested that APA in reservoirs was attached to particles larger than $0.4{\mu}m$, whereas in streams it existed in a dissolved form. There was a positive significant correlation between chlorophyll a concentration and APA in the photic zone. In the aphotic zone, APA correlated positively with the colony count of heterotrophic bacteria, but not with microscopic total bacterial counts.

Radiomodifying Potential of Panax ginseng in Liver of Swiss Albino Mice against Gamma Radiation

  • Sharma, Mukesh Kumar;Kumar, Madhu;Kumar, Ashok
    • Journal of Ginseng Research
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    • v.28 no.2
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    • pp.78-86
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    • 2004
  • Panax ginseng occupies an important role in the folk medicine of China, Korea and Japan. The present study was undertaken to determine the radioprotective efficacy of ginseng root extract in the liver of Swiss albino mice. The animals were divided into 4 groups. Group I-Only vehicle was administered. Group II-The animals received 10 mg/kg body weight ginseng root extract i.p. for 4 consecutive days. Group III-Animals were irradiated with 8Gy gamma radiation at the dose rate of 1.69 Gy/min at the distance of 80 ems. Group IV-Animals were given by ginseng root extract (10 mg/kg body weight) continuously for 4 days and on 4th day they were irradiated with 8 Gy gamma radiation after 30 min. The animals from above groups were autopsied on 1,3,7,14 and 30 days. Biochemical estimations of phosphatases (acid & alkaline), LDH (lactate dehydrogenase), LPO (lipid peroxidation) and GSH (reduced glutathione) in liver and SGOT (serum glutamate oxaloacetate transaminase), SGPT (serum glutamate pyruvate transaminase) and alkaline phosphatase in serum were done. In ginseng treated group acid phosphatase (ACP), alkaline phosphatase (ALP), LPO and LDH in liver and SGOT, SGPT and alkaline phosphatase in serum did not show any significant alteration. However, a significant increase in GSH content in liver was recorded. In irradiated group there was a significant increase in ACP, ALP and LPO content in liver and SGOT & SGPT in serum was noted. Whereas, a significant decrease was recorded in GSH and LDH activity in liver and alkaline phosphatase activity in serum. Pretreatment of ginseng with radiation significantly alters the biochemical parameters in liver and serum. A significant decline in ACP, ALP activity and LPO content in liver and SGOT and SGPT activity in serum was observed. However, a significant increase in GSH content and LDH activity in liver and ALP activity in serum was estimated. The present study suggests that pretreatment of ginseng before irradiation significantly protects the liver and maintains the enzyme activity.

The Effects of Palmijihwang-hwan (Baweidehuang-wan) and Obaeja (Galla Rhois) on Proliferation Activity of Alkaline Phosphatase and the Synthetic Ability of Protein in Osteoblast-like Cell Lines and Periodontal Ligament Fibroblasts (팔미지황환 및 오배자 추출물이 뼈모유사세포와 치주인대섬유모세포의 증식, Alkaline Phosphatase의 활성 및 단백질 합성능에 미치는 영향)

  • 김천종;안영민;안세영;두호경
    • The Journal of Korean Medicine
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    • v.24 no.3
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    • pp.35-44
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    • 2003
  • Objective : This study was performed to evaluate the effects of Palmijihwang-hwan (Baweidehuang-wan) and Obaeja (Galla Rhois) on the regeneration of periodontal tissue. Methods : In this study, we used MC3T3-El cells, such as osteoblast-like cell lines and human periodontal ligament fibroblasts, for experimental material. We separated each type of cells into a control group and an experimental group. In the control group, the cells were cultivated for 48 hours with distilled water and media which contained 10% fetal bovine serum (FBS) and penicillin (l00unit/ml)-streptomycin ($l00{\mu\textrm{g}}/ml$) at $37^{\circ}$ in 5% $CO_2$ gas. In the experimental group, the cells were cultivated for 48 hours with Palmijihwang-hwan extract and Obaeja extract (concentrations $1{\mu\textrm{g}}/ml,{\;}25{\mu\textrm{g}}/ml,{\;}50{\mu\textrm{g}}/ml$) under the same conditions as the control group. Investigating the regeneration of periodontal tissue was performed by evaluating proliferation, the activity of alkaline phosphatase and the synthetic ability of proteins using those cultivated cells by means of microculture tetrazolium (MTT) assay, alkaline phosphatase substrate kit and protein assay kit. Results : 1. In vitro, Palmijihwang-hwan extract increased the proliferation of MC3T3-El cells. 2. In vitro, Obaeja extract increased the activity of alkaline phosphatase and the synthetic ability of protein in MC3T3-El cells and human periodontal ligament fibroblasts depending on Obaeja extract's concentration. Conclusion : Obaeja extract can be developed as a subsidiary medicine for the regeneration of periodontal tissue. Further studies to evaluate the different concentrations the Obaeja extract and clinical trials in vivo are suggested.

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Studies on the activities of ALPase, ACPase, ATPase and accumulation of volutin granules upon growth phase in saccharomyces uvarum (Saccharomyces uvarum의 배양시기에 따른 ALPase, ACPase, ATPase 활성도와 volutin과립 축적량)

  • 이기성;최영길
    • Korean Journal of Microbiology
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    • v.23 no.2
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    • pp.90-100
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    • 1985
  • The present study was designed to investigate cellular regulation of phosphate metabolism between catabolically repressed and derepressed states in yeast (Saccharomyces uvarum). The activities of various phospatases and the contents of phosphate compounds were detected according to the culture phase and various phosphate concentrations. As the results, Saccharomyces uvarum derepressed many phosphate metabolizing enzymes such as alkaline phosphatase, acid phosphatase and ATPase more than ten fold simultaneously during catabolic repression (phospgate and sugar starvation). At the same state, the amounts of orthophosphate, nucleotidic labile phosphate and acid soluble polypgosphate were increased, compared to basal levels of normally cultivated cells. $Mg^{++}-stimulated$ type among all phospatases was appeared to have most of the enzyme activity. It could be postulated that $K^+ -stimulated$ alkaline phosphatase was directly or indirectly correlated with the synthesis of acid insoluble polyphosphate $Mg^{++}-stimulated$ phosphatase with the degradation of polyphosphates. In case of cultivation in the medium supplemented with sugar and phosphate (catabolic derepression), phospgatase activities except for alkaline phosphatase were decreased rapidly through the progressive batch culture, After 12 hrs culture, at early exponential phase, the cellular accumulation of acid insoluble polyphosphate increased about 5 fold, compared to those of the starved cells. Under catabolic repression, it could be postulated that intracellular phosphate metabolism was regulated by derepressions of phosphatases. The function of polyphosphate system was shown to compensate the ATP/ADP system as phosphate donor and energy source especially during catabolic repression.

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The Characterization and Activity Changes of Phosphatases in Amoeba sp. to the Light Stimuli and Its Response Pattern (광자극에 대한 Amoeba 의 반응양상과 phosphatase 의 특성 및 활성도 변화)

  • Chang, Nam-Kee;Chai-Seong Lim;Jin-Ho Bae
    • The Korean Journal of Ecology
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    • v.14 no.1
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    • pp.101-111
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    • 1991
  • Amoeba sp. was cultured under the light and the dark conditions, and the activity of phosphatases was investigated. There was a linear correlation between the early reaction time and the activity of phosphatases when phosphatases were incubated at 30℃. Then the activity of acid phosphatase was about 2 times higher than that of alkaline phosphatase. The activity of phosphatase was optimal at pH 5.0 in acidic part and at pH 8.0 in alkaline part, respectively. The optimal temperature of phosphatases was near the 40℃. The isozyme patterns of cytoplasmic acid phosphatase were compared with those of membraneous one. Both the isozyme patterns were shown to bo polymorphic on the polyacyamide gel, but different band patterns were observed in the isozymes of the cytoplasmic and the membraneous acid phosphatases. The number of Amoeba sp. under the light stimulus for 48 hours decreased negative exponentially from the illumination. The activity of acid and alkaline phosphatases under the illumination of light incresed 1.7 and 1.5 times higher, respectively, than the activity of those under the dark condition. This result apperars to be related to the mechanism of the autophosphorylation.

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