• Title/Summary/Keyword: Acid phosphatase-1

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Biochemical Markers for Osteosarcoma (골육종의 생화학적 표지자에 관한 연구)

  • Lee, Chang-Woo;Cho, Woo-Jin;Cho, Jae-Lim;Kim, Tai-Seung;Whang, Kuhn-Sung
    • The Journal of the Korean bone and joint tumor society
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    • v.7 no.2
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    • pp.41-50
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    • 2001
  • Purpose : To investigate biochemical markers for osteosarcoma, activities of deoxyribocuclease(DNase), ribonuclease(RNase), 5'-nucleotidase, alkaline phosphatase and amylase were determined in the osteosarcoma tissue and serum of patients with osteosarcoma. Also studied were DNase, RNase in osteosarcoma tissue, isolating the enzymes from the sarcoma tissue and investigating the sarcoma specific enzymes. Materials and Methods : The experimental tissue and serum were obtained from twelve patients with osteosarcoma. The control group were obtained from the normal healthy tissue of the same patients. The tissue were centrifugalized to obtain extracts. The extracts were analized for the estimation of nucleic acid, protein contents and enzyme activities. And then each enzymes were isolated and analized by DEAE-cellulose chromatography and estimated for activities. Result : Activities of acid DNase, RNase, 5'-nucleotidase and alkaline phosphatase were significantly increased in osteosarcoma tissue. Neutral RNase in osteosarcoma tissue was shown to bo highly active, exhibiting secretory form of RNase inhibitor associated with the RNase was also increased. In the serum of patients with osteosarcoma, RNase activity was significantly increased. DEAE-cellulose column chromatographical analysis revealed that acid DNase was isolated as a single enzyme and neutral RNase as five isozymes in osteosarcoma tissue. Conclusion : The results indicated that combination of these enzymes could be used as markers for osteosarcoma. The results indicated that acid DNase and neutral RNase might play a role in genesis of sarcoma and suppression of sarcoma.

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Dedifferentiation Correlates with the Expression of Lysosomal Acid Phosphatase in the Limb Regenerates of Mexican Axolotl (멕시코산 엑소로틀 다리 재생조직의 탈분화와 리소솜 산성탈인산화효소의 발현)

  • Seo, Kwang-Seok;Park, Sook-Kyung;Ju, Bong-Gun;Jeon, Sang-Hak;Kim, Won-Sun
    • Development and Reproduction
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    • v.2 no.1
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    • pp.53-62
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    • 1998
  • The lysosomal acid hydrolases including lysosomal acid phosphatase (LAP) are believed to play an important role in intracellular and extracellular degradation. LAP was reported to increase its activity in dedifferentiation stage during urodele limb regeneration. In the paresent study, LAP localization in the Mexican axolotl (Ambystoma mexicanum) limb regenerates was investigated by immunohistochemistry. LAP immunoreactivity with monoclonal antibody against Korean salamander (Hynobius leehii) LAP was observed mainly in the wound epidermis, blastema cells, muscle, and cartilage which were under dedifferentiation process in axolotl limb regenerates. Moreover, LAP immunoreactivity increased gradually during the early phase of lib regeneration and reached the peak level at dedifferentiation stage. However, as redifferentiation begans, LAP immunoreactivity decreased slowly to the basal level. Retinoic acid (RA) which is known to induce skeleton pattern duplication in regenerating urodele limb appears to enhance LAP immunoreactivity. In the RA-treate limg regenerates, LAP immunoreactivity was higher than in the normal regenerates. In addition, the LAP expression period was more extended in the RA treated regenerates than in the normal regenerates. These results suggest that RA is involved in the extension of dedifferentiation state in RA-treated limb regenerate.

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2D-QSAR and HQSAR on the Inhibition Activity of Protein Tyrosine Phosphatase 1B with Oleanolic Acid Analogues

  • Chung, Young-Ho;Jang, Seok-Chan;Kim, Sang-Jin;Sung, Nack-Do
    • Journal of Applied Biological Chemistry
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    • v.50 no.2
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    • pp.52-57
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    • 2007
  • Quantitative structure-activity relationships (QSARs) on the inhibition activities by oleanolic acid analogues (1-19) as a potent inhibitor against protein tyrosine phosphatase-1B were studied quantitatively using 2D-QSAR and HQSAR methodologies. The inhibition activity was dependent on the variations of $R_{4-}$substituent, and as shown in 2D-QSAR model ($r^2=0.928$), it has a tendency to increase as the negative Randic Indice (RI) goes up. The size of the molecular fragments used in HQSAR varied from five to eight. The fragment distinctions had the best statistic value, whose predictability is $q^2=0.785$ and correlation coefficient is $r^2=0.970$, on condition of connections. From the atomic contribution maps, the factor that contributes to the inhibition activities is the $C_{15}{\sim}C_{17}$ bond in the D ring. From the analysis result of these two the models, the structural distinctions and descriptors that contribute to the inhibition activities were obtained.

Characterization and Cloning of a Phytase from Escherichia coli WC7. (Escherichia coli WC7가 생산하는 Phytase의 효소특성과 그 유전자의 클로닝)

  • 최원찬;오병철;김형권;강선철;오태광
    • Microbiology and Biotechnology Letters
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    • v.30 no.1
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    • pp.1-7
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    • 2002
  • Phytase from Escherichia coli WC7 was purified from cell extracts and its molecular mass was estimated to be 45 kDa by SDS-PAGE. Its optimum temperature and pH for phytate hydrolysis was 6$0^{\circ}C$ and pH 5.0, respectively. The enzyme was stable up to 6$0^{\circ}C$ and over broad pH range (pH 2-12). The enzyme had higher affinity for sodium phytate than p-nitrophenylphosphate (pNPP). That is, the apparent Km value for sodium phytate and pNPP were $0.15\pm$0.02 mM and 2.82$\pm$0.05 mM, respectively. The gene encoding the phytase was cloned in E. coli XL1-Blue. Sequence analysis showed an open reading frame of 1241 Up encoding a signal peptide (22 aa) and a mature enzyme (410 aa). WC7 phytase was expressed up to 17.5 U/ml in the transformed E. coli XL1-Blue/pUEP, which was 23-fold higher than the activity from wild strain.

Effect of Electroacupuncture on the Regeneration of Injured Peripheral Nerve (손상된 말초신경의 재생에 미치는 전자침의 효과)

  • Deung, Young-Kun;Kim, Dong-Heui;Kang, Song-Jian
    • Applied Microscopy
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    • v.32 no.1
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    • pp.1-8
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    • 2002
  • To confirm the effect of electroacupuncture on the regeneration of injured peripheral nerve, the change of evoked potential in the sciatic nerve, the change of enzyme activity in the spinal cord, and morphological change of injured sciatic nerve were examined comparatively in acupuncture group (AG) and control group (CG) after sciatic nerve of guinea pig was injured by purpose. The value of evoked potential after injury of the sciatic nerve was increased in both AG and CG, but the increase rate of that was higher in AG than CG. Acid phosphatase activity of the spinal cord was increased in 1CG and 2AG, but shown are tendency to return to the normal state as time went by. Ultrastructural recovering rate of the injured sciatic nerve was higher in AG than CG. Also, there was developed only adipose tissue in sciatic nerve of AG. As mentioned above, the effect of electroacupuncture on the regeneration of injured peripheral nerve was confirmed experimentally by change of evoked potential, acid phosphatase and ultrastructure. Especially, the effect of electroacupuncture was appeared clearly in an early stage than other treatment stages.

Purification and Characterization of Protein Phosphatase 2A from Petals of the Tulip Tulipa gesnerina

  • Azad, Md. Abul Kalam;Sawa, Yoshihiro;Ishikawa, Takahiro;Shibata, Hitoshi
    • BMB Reports
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    • v.39 no.6
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    • pp.671-676
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    • 2006
  • The holoenzyme of protein phosphatase (PP) from tulip petals was purified by using hydrophobic interaction, anion exchange and microcystin affinity chromatography to analyze activity towards p-nitrophenyl phosphate (p-NPP). The catalytic subunit of PP was released from its endogenous regulatory subunits by ethanol precipitation and further purified. Both preparations were characterized by immunological and biochemical approaches to be PP2A. On SDS-PAGE, the final purified holoenzyme preparation showed three protein bands estimated at 38, 65, and 75 kDa while the free catalytic subunit preparation showed only the 38 kDa protein. In both preparations, the 38 kDa protein was identified immunologically as the catalytic subunit of PP2A by using a monoclonal antibody against the PP2A catalytic subunit. The final 623- and 748-fold purified holoenzyme and the free catalytic preparations, respectively, exhibited high sensitivity to inhibition by 1 nM okadaic acid when activity was measured with p-NPP. The holoenzyme displayed higher stimulation in the presence of ammonium sulfate than the free catalytic subunit did by protamine, thereby suggesting different enzymatic behaviors.

Effects of bisphosphonate and indomethacin on alveolar bone remodeling in rats (Bisphosphonate와 Indomethacin이 백서 치조골의 골개조에 미치는 영향)

  • Cho, Myeong-Sook;Kim, Jong-Chul
    • The korean journal of orthodontics
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    • v.26 no.2 s.55
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    • pp.163-174
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    • 1996
  • The purpose of this study was to examine the effects of bisphosphonate and indomethacin, blockers of bone resorption with different mechanisms, on alveolar bone remodeling. Male rats were divided into control, bisphosphonate and indomethacin groups, and then each group was divided info an experimental side and a control side according to the force application. Bisphosphonate(6.3mg/kg,$2.52x10^{-2}mol/L$) and indomethacin (9mg/kg, $2.52x10^{-2}mol/L$) were injected 6 hours and 1 hour before or 24 hours after the force application. The rats were killed 72 hours after the force application and histologic examination was perfomed. The values of serum acid phosphatase and lactate dehydrogenase were also measured in the control md experimental groups treated with bisphosphonate or indomethacin 1 hour before the force application. In the experimental side, the least number of osteoclasts was noted in the groups treated 1 hour before the force application with indomethacin or bisphosphonate, while there were no differences between the control and the groups treated with drugs 6 hours before or 24 hours after the force application. In the control side, the number of osteoclasts was not inecreased with no differences among the groups. Histologic examination revealed a severe alveolar bone resorption in the control group and the groups treated with indomethacin 6 hours before or 24 hours after the force application. Indomethacin treatment 1 hour before the force application and bisphosphonate treatment at any time significantly attenuated the bone resorption. Electron microscopically, ruffled border and clear zone of osteoclasts were observed in the control and indomethacin groups, while some osteoclasts were detached from the bone surface and exhibited dull cellular projections in the bisphosphonate groups. The bisphosphonate and indomethacin groups showed lower values of acid phosphatase and lactate dehydrogenase than the control group. The acid phosphatase value in the bisphosphonate group was lower than that in the indomethacin group, whereas there was no difference in the lactate dehydrogenase value between the groups. These results suggest that bisphosphonate reduces the activity of osteoclasts as well as the number of osteoclasts and that indomethacin reduces the number of osteoclasts without affecting the activity of osteoclasts. Bisphosphonate has a larger inhibitory effect on bone resorption md thus less limitation in the application time than indomethacin.

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