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Biochemical Changes in the Hemolymph of the Larvae of Thecodiplosis japonensis Uchi. et Inouye (솔잎혹파리 유충 체액의 생화학적 변화)

  • Lee Kyung-Ro;Lee Jong-Jin
    • Korean journal of applied entomology
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    • v.15 no.4 s.29
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    • pp.169-178
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    • 1976
  • The concentration of amino acids, total nitrogen, trehalose, lipids and the activities of respiratory, acid$\cdot$alkaline phosphatase, glutamic oxalozcetic transaminase and glutamic pyruvic transaminase during larval stage in Pine leaf gall midge, Thecodiplosis janensis Uchi. et Inouye were measured using Paper chromatographic method, micro-Kjeldahl method, Thin layer chromatographic method, Warburg's manometric method, Bessey-Lowry method and Reitman-Frankel method, respectively. Healthy specimens )yore chosen as samples of each larval stages; alrva in gall and larva in soil. Amino acids present in the alcoholic extracts were alanine, glutamic acid, glycine, histidine, methionine, proline, threonine, tryptophan and valine. The total nitrogen concentration reached to 31.348mg/g during the larva in gall and the larval stage in soil of the value was decreased to 29.027mg/g. The hemolymph sugar, trehalose value for larva in soil was about two times of the value for larva in gall. Total lipid, phospholipid,monoacylglycerol, triacylglycerol, sterol, free fatty acid and ester cholesterol were identified at larval stages in gall and soil. Triacylglycerol concentration reached high level in contrast with other lipid contents during larvae in gall and larva in soil. Free fatty acid, sterol except decreased lipids during larval stage in soil. Endogenous respiration, succinate of respiratory activities decreased at larval stage in soil compare with larva in gall. The activities of acid phosphatase decreased larval stage in soil but the activities of alkaline phosphatase increased remarkably. The activities of glutamic oxaloacetic transaminase and glutamic pyruvic transaminase reached high level of the larva in gall.

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EFFECTS OF SEVERAL CYTOKINES ON THE FUNCTIONS OF FETAL RAT OSTEOBLAST-LIKE CELLS IN VITRO

  • Han, Hee-Sung;Kim, Jung-Keun;Chang, Young-IL
    • The korean journal of orthodontics
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    • v.25 no.6 s.53
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    • pp.689-696
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    • 1995
  • Effects of several cytokines($IL-1{\beta},\;TNF_{\alpha},\;and\;IFN_{\gamma}$) have been examined on fetal rat osteoblast-like cells. To investigate whether cytokines play direct causal roles in production of lysosomal enzyme, fetal rat osteoblast-like cells were treated with $IL-1{\beta},\;TNF_{\alpha},\;and\;IFN_{\gamma}$, respectively or combined. And acid phosphatase was determined by biochemical method. Alkaline phosphatase was assayed to determine the effects of $IL-1{\beta},\;TNF_{\alpha},\;and\;IFN_{\gamma}$ on the expression of this enzyme. And also experiment of calcified nodule formation was performed to assess the effects of cytokines on the bone-forming activity of osteoblast-like cells in vitro. Acid phosphatase activity was significantly increased by the addition of $IL-1{\beta}\;and\;TNF_{\alpha}$, whereas decreased by $IFN_{\gamma}$. However, no significant change:: in alkaline phosphatase activity was observed when the osteoblast-like cells were treated with $IL-1{\beta}\;and\;TNF_{\alpha}$. Interestingly, $IFN_{\gamma}$ showed stimulatory effect on alkaline phosphatase activity. The number of calcified nodules was decreased by treatment of cultures with 1 ng/ml $IL-1{\beta},\;20\;ng/ml\;TNF_{\alpha}$, and 500 u/ml $IFN_{\gamma}$ continuously for 21 days, while considerable number of calcified nodules were formed in control group of osteoblast-like cell in culture for 21 days. These results seem to suggest that cytokines may play crucial roles in bone remodeling through the direct action on the osteoblast-like cell.

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A Comparative Study on the Effects of Polygoni Radix and Cynanchi Radix on Rat Livers Intoxicated with Carbon Tetrachloride (백서(白鼠) 간조직(肝組織)에 미치는 적하수오(赤何首烏)와 백하수오(白何首烏)의 효능에 관한 비교(比較) 연구(硏究))

  • Shin, Min-Kyo
    • Korean Journal of Pharmacognosy
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    • v.16 no.2
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    • pp.81-92
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    • 1985
  • The Polygoni Radix and Cynanchi Radix have been used to potentiate the liver functions in clinic of Oriental Medicine. The water extracts of Polygoni Radix and Cynanchi Radix were administered orally to rats intoxicated with carbon tetrachloride and then this experiment have been performed by observing liver fatty degeneration and activities of enzymes such as cytochrome oxidase (CYO), adenosine triphosphatase (ATP), acid phosphatase (ACP), lactate dehydrogenase (LDH) and alkaline phosphatase (ALP). By oral administration of water extracts of the radices between 1 and 10 days, the following results were obtained. 1. The group given Cynanchi Radix extract showed recovery of the fat liver in 4 days, whereas that given Polygoni Radix extract did the recovery in 8 days. 2. In cytochrome oxidase activity, the group given Cynanchi extract showed normal activity in 6 days, whereas that given Polygoni Radix extract did the activity in 8 days. 3. In adenosine triphosphatase activity, the groups given Cynanchi Radix and Polygoni Radix extracts showed normal activities in 2 and 8 days, respectively. 4. In acid phosphatase activity, the groups given Cynanchi Radix and Polygoni Radix extracts showed recovery of the activities in 2 and 4 days, respectively. 5. In lactate dehydrogenase activity, the group given Cynanchi Radix and Polygoni Radix extracts showed recovery of the activities in 6 and 10 days, respectively. 6. In alkaline phosphatase activity, the group given Cynanchi Radix extract showed normal activity in 2 days, whereas that given Polygoni Radix extract showed slight recovery between 4 and 6 days followed by decrease of the activity in 8 and 10 days. From the above-mentioned results, it was found that both of the water extracts of Polygoni and Cynanchi Radix possessed the recovery action of liver function as intoxicated with carbon tetrachloride in rats. It is also noted that the extract of Cynanchi Radix showed more potent activity than that of Polygoni Radix.

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Ultrastructural and Cytochemical Studies on the Endometrial Surface Epithelial Cells of Guinea Pig During Estrous Cycle (發情週期에 따른 Guinea Pig의 子宮內膜 表層上皮細胞의 微細構造 및 細胞化學的 硏究)

  • Park, Choon K.;Kwan H. You;Young K. Deung;Lee, Choon K.;Ho S. Chung
    • The Korean Journal of Zoology
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    • v.25 no.1
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    • pp.9-28
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    • 1982
  • Cyclical changes in the fine structures of the surface epithelial, stroma and glandular cells of guinea pig endometrium during the estrous cycle were studied by transmission and scanning electron microscopy. Cytochemical studies were made in order to investigate the ultrastructural localization of the acid phosphatase, alkaline phosphatase and ATPase in these cells. The results obtained are as follows: 1. The endometrial surface epithelium was pseudostratified columnar during estrus and meterstrus, and simple columnar during proestrus and diestrus. The characteristic features observed in these cells include increased nucleocytoplasmic ratio at proestrus, elongated shapes of both the nucleus and the entire cell, increased volume of the cytoplasm and cytoplasmic bulding into the lumen during estrus, and smaller surface epithelial cells during metestrus. 2. In the cytoplasm of surface epithelial cells, the numbers of mitochondria and free ribosomes were increased, and rough endoplasmic reticulum and Golgi complex appeared during estrus, and the degenerated cells, lipid droplets, multilamellated bodies and lysosomes appeared during diestrus. 3. During estrus, scanning electron microscopic observations of endometrial surface showed a regular arrangement with polygonal outlines of epithelial cells, distinct intercellular border, and bulged surface into the lumen, whereas flat surface and indistinct cell border were characteristic during meterstrus and diestrus. 4. Microvilli which aligned on the surface were longer and most abundant during estrus while short and aparse during other phases. 5. Cytochemical studies indicated that during metestrus acid phosphatase activities were localized in the microvilli and vacuoles, and alkaline phosphatase activities were significant around luminal surface and lateral cell membrane in the surface epithelial cells. ATPase activities were present on the microvilli and cell membrane during proestrus and estrus.

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CHANGES OF SERUM ALKALINE PHOSPHATASE AFTER ENUCLEATION OF CYSTS IN THE JAWS (낭종 수술 전후에 있어서 혈중 Alkaline Phosphatase의 변화에 대한 연구)

  • Eune, Jung-Ju;Lee, Eui-Seok;Rim, Jae-Suk;Jang, Hyon-Seok;Woo, Hyon-Il
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.31 no.5
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    • pp.417-421
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    • 2005
  • This study was to analyze the changes of levels of alkaline phosphatase before and after enucleation of jaw cysts combined with bone grafting, and to evaluate biochemically the effectiveness of the early detection of bone healing and infection as a prognostic marker. Eighteen patients (13 males, 5 females) with cystic lesions of the jaws were divided into two groups. The bone graft group underwent enucleation and bone graft. The control group underwent only enucleation. Both groups were measured levels of ALP before surgery, and plus-minus 4 weeks postoperatively. The more discriminating results were obtained in the bone graft group. The results were as follows : 1. Levels of ALP after enucleation of jaw cysts were decreased in all patients with and without bone graft. 2. The bone graft group showed more marked decrease in variation of levels of ALP than the control group.(p=0.008) This should be considered as a result of increased osteoblastic activity and new bone formation. 3. Such variation could be used as a prognostic marker for bone healing after cyst operation. In the cost/benefit ratio, measurement of ALP activity could be useful as a convenient procedure in routine clinical practice.

A Study on the Effect of Physical Stimuli on Bone Cell Differentiation Using a Hybrid Bioreactor (Hybrid Bioreactor를 이용한 물리적 자극에 대한 세포반응 연구)

  • 이창양;최귀원;홍대희
    • Journal of Biomedical Engineering Research
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    • v.25 no.4
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    • pp.309-314
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    • 2004
  • in this study, hybrid bioreactor was used to apply physical stimuli in cell culture. Effect of the applied physical stimuli on the growth and differentiation of MC3T3-El cell in a three-dimensional Chitosan scaffold were studied by using the hybrid bioreactor. The hybrid bioreactor for physical stimulus was specially designed to apply uniaxial cyclic compressive and shear strain. Physical stimulus was applied over a period of 14 days with 150 cycles per day at a frequency of 0.5Hz. Strain magnitude was 2.5% of the scaffold size. Control group and physically stimulated group of the MC3T3-El tell were incubated and harvested at the indicated times (Day 6, 8, 10, 12, 14). The total amount of protein, which obtained information of cell growth, was determined by Lowey method. Alkaline phosphatase activity was examined by ELISA. Physically stimulated group using the hybrid bioreactor was increased in alkaline phosphatase activity comparing with control group. The nodule formation and calcium deposit of the physical stimuli group which resulted in cell differentiation was faster than that of control group.

Effect of Atrial Natriuretic Peptide on the Proliferation and Activity of Osteoblastic Cells

  • Lee, Jong-Ryeul;Ko, Seon-Yle;Kim, Jung-Keun;Kim, Se-Won
    • The Korean Journal of Physiology and Pharmacology
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    • v.4 no.4
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    • pp.283-289
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    • 2000
  • Natriuretic peptides comprise a family of three structurally related peptides; atrial natriuretic peptide (ANP), brain natriuretic peptide (BNP), and C-type natriuretic peptide (CNP). The present study was performed to investigate the effect of ANP on the proliferation and activity of ROS17/2.8 and HOS cells which are well-characterized osteoblastic cell lines. ANP dose-dependently decreased the number of ROS17/2.8 and HOS cells after 48-hour treatment. ANP generally increased the alkaline phosphatase activity of ROS17/2.8 and HOS cells after 48 hr treatment, regardless of the fact that basal activity of alkaline phosphatase was much lower in HOS cells compared to that of ROS17/1.8 cells. ANP increased the NBT reduction by ROS17/2.8 and HOS cells. ANP showed the variable but no significant effect on the nitric oxide production by ROS17/2.8 and HOS cells. ROS17/2.8 and HOS cells produced and secreted gelatinase into culture medium, and this enzyme was thought to be the gelatinase A type with the molecular weight determination. The gelatinase activity produced by ROS17/2.8 cells was increased by the treatment of ANP. However, the enzyme activity was not affected by ANP treatment in the HOS cell culture. In summary, ANP decreased the proliferation and increased the alkaline phosphatase activity and NBT reduction of osteoblasts. These results indicate that ANP is one of the important regulators of bone metabolism.

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Expression of a Functional zipFv Antibody Fragment and Its Fusions with Alkaline Phosphatase in the Cytoplasm of an Escherichia coli

  • Hur, Byung-Ung;Choi, Hyo-Jung;Yoon, Jae-Bong;Cha, Sang-Hoon
    • IMMUNE NETWORK
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    • v.10 no.2
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    • pp.35-45
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    • 2010
  • Background: Expression of recombinant antibodies and their derivatives fused with other functional molecules such as alkaline phosphatase in Escherichia coli is important in the development of molecular diagnostic reagents for biomedical research. Methods: We investigated the possibility of applying a well-known Fos-Jun zipper to dimerize $V_H$ and $V_L$ fragments originated from the Fab clone (SP 112) that recognizes pyruvate dehydrogenase complex-E2 (PDC-E2), and demonstrated that the functional zipFv-112 and its alkaline phosphatase fusion molecules (zipFv-AP) can be produced in the cytoplasm of Origami(DE3) trxB gor mutant E. coli strain. Results: The zipFv-AP fusion molecules exhibited higher antigen-binding signals than the zipFv up to a 10-fold under the same experimental conditions. However, conformation of the zipFv-AP seemed to be influenced by the location of an AP domain at the C-terminus of $V_H$ or $V_L$ domain [zipFv-112(H-AP) or zipFv-112(L-AP)], and inclusion of an AraC DNA binding domain at the C-terminus of VH of the zipFv-112(L-AP), termed zipFv-112(H-AD/L-AP), was also beneficial. Cytoplasmic co-expression of disulfide-binding isomerase C (DsbC) helped proper folding of the zipFv-112(H-AD/L-AP) but not significantly. Conclusion: We believe that our zipFv constructs may serve as an excellent antibody format bi-functional antibody fragments that can be produced stably in the cytoplasm of E. coli.

Inhibitory Effect of Retinoids on Alkaline Phosphatase Isoenzymes Activity in Human Serum

  • Kim, Seung Hee;Moon, Ki-Young
    • Biomedical Science Letters
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    • v.23 no.3
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    • pp.230-237
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    • 2017
  • Changes in the activity of alkaline phosphatase (ALP) isoenzymes and isoforms in human serum have a major diagnostic value, therefore the regulation of ALP activities is a valuable target for therapeutic interventions. To assess the pharmacological activity of retinoids, i.e., all-trans retinoic acid and 13-cis retinoic acid, their tissue-specific inhibitory effect on human serum ALP activity was elucidated by chemical inhibition methods, heat-sensitive inactivation, and wheat-germ lectin precipitation test. Retinoids showed significant inhibition of the total ALP activity in human serum at a concentration of 5 mM. All-trans retinoic acid (5 mM) and 13-cis retinoic acid (5 mM) inhibited ALP activities by up to 12% and 15%, respectively, compared to that by guanidine hydrochloride (200 mM). L-phenylalanine (100 mM) and urea (30 mM) had no further inhibitory effect on ALP activities in human serum pretreated with retinoids (5 mM). Retinoids significantly inhibited ALP activities by up to 20% compared with that of tetramisole (30 mM). The ALP activities in retinoid-pretreated serum remained unchanged after the heat inactivation process. These results suggest that retinoids are inhibitors of the intestinal ALP isoenzyme. Remarkably, retinoids revealed potent inhibitory activities against ALP in wheat-germ lectin precipitant serum, indicating that they also function as inhibitors of the bone ALP isoform. The results show that retinoids inhibit the specific tissue-derived human serum ALP activities, moreover, the inhibitory effect of retinoids against bone ALP activity suggests their clinical utility as monitoring and prevention of metastasis of bone cancer.