• Title/Summary/Keyword: Alkaline phosphatase

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A Study of Alkaline Phosphatase Activity on the Preimplantation Mouse Embryos (초기 흰쥐 배아의 발생단계에 있어서의 Alkaline Phosphatase의 활성에 관한 연구)

  • Cho, Wan-Kyoo;Lee, Chung-Choo;Kim, Hee-Kwon
    • The Korean Journal of Zoology
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    • v.27 no.1
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    • pp.1-12
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    • 1984
  • In order to investigate the alkaline phosphtase activities in the mouse oocytes in matuation and preimplantation embryos in developing in culture, the enzyme activities were measured by means of biochemical method. The in vitro effect of levamisole which is known as an inhibitor of the lakaline phosphatase was also observed on the oocyte in maturation and the embryos in early embryogenesis. The results obtained were as follows: The enzyme activity was not detected in the embryos unitl the stage of 4-cell, but it appeared first in the 4-cell embryos and the level of the activity was steady through up to the blastocyst. Levamisole inhibited the alkaline phosphatase activity in the blastocyst, and the activity decreased by almost 70% at 10 mM and 50% at 1 mM as compared with the control. In addition, levamisole inhibited completely the formation of polar body by the oocytes. and induced degeneration of the preimplantation embryos at the dose of 0.5 mM or higher.

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Localization and isozyme patterns of phosphatase in Fibricola seoulensis (Fibricola seoulensis에서 phosphatase의 분포와 동위효소유형)

  • 김홍자;김창환
    • Parasites, Hosts and Diseases
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    • v.31 no.4
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    • pp.353-362
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    • 1993
  • The present study was carried out to investigate the localization and isozyme patterns of acid phosphatase and alkaline phosphatase in metacercariae and in adults of F. seoulensis by enzyme-histochemistry method and electrophoresis. Acidphosphatase showed a strong activity at pH 5 in the intestinal caecum of adults, but showed no reactions in the nonsubstrate control and in the inhibitor-treated control. Alkaline phosphatase showed a strong activity at pH 8 in the intestinal caecum and the tribocytic organ of adults, and in the intestinal caecum and in the genital anlagen of metacercariae. In non-denature PAGE, ten bands of protein fraction from the extracts of metacercariae and twenty-two bands from adults were detected. In denature PAGE, two protein bands having molecular weights of 192 kDa and 123 kDa were detected in the metacercariae, but absent from adult stage. In adults, protein fractions of 27.5 kDa, 24.5 kDa, 21.4 kDa, 18 kDa, 16 kDa and 15 kDa were detected. In non-denature PAGE, isozymes of acid phosphatase showed the most strong activity at pH 5, whereas no activity was shown at pH 2 and pH 7. One isozyme 85 kDa, 73 kDa and 62 kDa) in adults.

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Application of alkaline phosphatase staining of cytology specimen for differential diagnosis of canine osteosarcoma (세포 검사시료에서 alkaline phosphatase 염색법을 활용한 개 골육종의 감별 진단)

  • Park, Byoung-Yong;Park, Chul;Cho, Ho-Seong
    • Korean Journal of Veterinary Service
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    • v.34 no.4
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    • pp.397-401
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    • 2011
  • Aspiration of lytic bone lesions is an excellent diagnostic test in the initial evaluation of primary bone tumor. However, cytologically, it can be difficult to differentiate osteosarcoma (OSA) from other bone neoplasms, including fibrosarcoma, chondrosarcoma, synovial cell sarcoma, malignant fibrous histiocytoma and malignant peripheral nerve sheath tumor. The purpose of this study is to introduce alkaline phosphatase (ALP) staining to differentiate OSA from other mesenchymal tumors. Tumors actively producing bone are specifically positive for ALP staining. Unstained, cytologic specimens were incubated for 10 minutes with nitroblue tetrazolium chloride/5-bromo-4-chloro-3-indolyl phosphate toluidine salt-phosphatase substrate. Among 20 cases of cytology specimen, 14 were positive for ALP staining and histopathology, 6 were negative for ALP staining and histopathology. ALP staining was 100% sensitive and specificity for the diagnosis of OSA. Aspirate cytology with ALP staining was a simple, fast, safe and accurate diagnostic test for the evaluation of suspected OSA lesions in dogs.

Histochemistry of the Developing Chick Respiratory Tract (鷄胎呼吸氣道의 組織化學的 硏究)

  • Hah, Jae Chung
    • The Korean Journal of Zoology
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    • v.15 no.4
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    • pp.183-191
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    • 1972
  • A histochemical study of the respiratory tract in developing chick was done to demonstrate PAS-postivie materials, ribonucleic acid, phospholipid, and alkaline phosphatase activity. Following results are obtained: 1. The alkaline phosphatase activity was found to be high before the appearance of cartilage in mesenchyme surrounding the tracheal epithelium. The enzyme activity declined after the cartilage formation, followed by the restricted activity in epithelium in the postembryonic stage. 2. A moderate positive reaction of ribonucleic acid was found in the cytoplasm of the epithelium and undifferentiated mesenchyme. As the cartilage grew differentiated the reaction of ribonucleic acid was found to disappear in the mesenchyme surrounding the epithelim, but the cytoplasm of the glands showed a moderate positive reaction. 3. Goblet cells of the mucosa and glandular cells showed highly positive reaction, but the basement membrane exhibited slightly positive PAS-reaction. 4. Epithelial cells of the mucosa showed a weak to moderate reaction. However, the epithelia of bronchiol and alveoli in the differentiating period and glandular cells showed a strong positive reaction in Baker's hematein test.

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Distribution and Some Properties of the Alkaline Phosphatase from the Midgut of the Earthworm, Eisenia andrei (지렁이 중장에서 분리한 Alkaline Phophatase의 분포와 특성에 관한 연구)

  • 박희우;조성진;조표연;이명식;이규석;박순철
    • The Korean Journal of Soil Zoology
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    • v.5 no.1
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    • pp.39-46
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    • 2000
  • The distribution and some properties of alkaline phosphatase (ALP) were investigated in the midgut of the earthworm, Eisenia andrei. The ALP activity appeared to be highly polarized toward the luminal side of epithelium, with minor ALP activity in chloragogue tissue. The epithelial and chloragogeneous tissues contained approximately 85 and 15% of total intestinal ALP activity, respectively. The optimal temperature was approximately 37$^{circ}C$ and isoelectricpoint was estimated to be 4. The treatment of neuraminidase and PtdIns-PLC failed to change the migration rate of ALPs. Also, these ALPs appeared to have a wide range of substrate specificity. The relationship between the properties and physiological significances of the midgut ALPs in Eisenia andrei was discussed.

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Changes in Phosphatase Activities of Mouse Epididymal Spermatozoa during Maturation (생쥐 부정소 정자의 성숙과정에서 Phosphatase 활성도 변화)

  • 김문규;윤현수;김종흡;김성례
    • The Korean Journal of Zoology
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    • v.33 no.1
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    • pp.70-77
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    • 1990
  • The change of phosphatase activities of the epididymal spermatozoa has been examined during epididymal maturation in mouse. The quantitative analysis of phQsphatase activities have been carried out using the method modified by Emst(1975). The results of experiment were summarized as the followings. Total protein of the caput epididyrnal spermatozoa(CPS) was measured as 59.1 $\pm$8.4(mg/10 9 spermatozoa), and that of the cauda epididymal spermatozoa(CDS) was 14.0$\pm$12.3(mg/10 9 spermatozoa). When phosphatase activities of the CDS in basic reaction medium were 29.2% in alkaline phosphatase, 44.9% in ATPse and 53.8% in acid phosphatase. The activities were eminently decreased in all CDS in contrast to those of CPS. The alkaline phosphatase and ATPase activities of K+ -dependent were decreased in CDS when compared with caput epididymal spermatozoa, and alkaline phosphatase, ATPase and acid phosphatase activities of $Ca^2$+ -dependent were increased in homogenized spermatozoa when compared with intact spermatozoa. From these results, it may be concluded that the decrease of phosphatases activities in spermatozoa during epididymal maturation may play some significant roles in acquiring fertilizing capability.

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Effects of Sodium Butyrate on the Biosynthesis of Sphingolipids in HT29, a Human Colon Cancer Cell Line (Sodium Butyrate 처리가 대장암 세포주인 HT29 Cell의 Sphingolipid 생합성에 미치는 영향)

  • 김희숙
    • Journal of Life Science
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    • v.9 no.2
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    • pp.160-168
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    • 1999
  • Butyrate is one of the short-chain fatty acids that are present in the colon of mammals in millimolar concentration as a result of microbial anaerobic fermentation of dietary fiber, undigested starch, and proteins. In this study, sodium butyrate was examined in HT29 cell, human colonic cancer cell line, on cell viability, alkaline phosphatase activity, PLC-${\gamma}$1 expression and complex sphingolipid biosynthesis. Treatment with butyrate showed that the decrease of cell adhesion and viability was time-dependent. Sodium butyrate also induced to increase the activity of alkaline phosphatase which is a differentiation marker enzyme and decrease the expression of PLC-${\gamma}$1. Biosynthesis of sphingomyelin and galactosylceramide by butyrate treatment were decreased so fast but ceramide was increased 680dpm/mg protein% more than untreated group on first day and then decreased fast. In addition, acid ceramidase and neutral ceramidase activity were inhibited early stage by sodium butyrate. These results suggest that sodium butyrate causes cell differentiation or cell growth arrest of HT29 cell accompanied by early increase of ceramide content and alkaline phosphatase activity and decrease of galactosylceramide content and PLC-r1 expression.

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The Effect of Fluoride and Aluminum on Bone Turnover in Mouse Calvarial Culture

  • Ahn, Hye-Won
    • Toxicological Research
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    • v.14 no.2
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    • pp.163-169
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    • 1998
  • Fluoride (F), over a narrow concentration range, increases bone formation. Aluminum (Ai) too is biphasic in its action on bone, being mitogenic at very low levels and inhibitory at higher levels. Both F and Al are present in finished drinking water where the chemical interaction of these two agents is well characterized. F and AI, given individually, accumulate preferentially in bone. In addition. in vivo studies have shown that F causes the co-accumulation of Al in bone. Thus, it was necessary to determine the interactive effect of these two agents on bone mitogenesis. Calvaria were obtained from neonatal CD-1 mice and cultured with various concentrations of F (0.05~19 ppm) as NaF, Al (2 ppb~2 ppm) as $AlCl_3$ , or F and Al for 3 days at $37^{\circ}C$ on a rotating roller drum. Alkaline phosphatase activity in calvaria and $\beta$-glucuronidase activity in culture medium were determined as a measures of bone turnover. Alkaline phosphatase activity in calvaria was significantly increased by F (0.05~2 ppm) treatment and $\beta$-glucuronidase activity was slightly increased in the culture medium of calvaria treated with 0.3 ppm Al. The combination of 19 ppm F and 0.3 ppm Al increased alkaline phosphatase activity in calvaria, but did not affect $\beta$-glucuronidase activity, suggesting the interactive effect of fluoride and aluminum on bone turnover.

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AN EXPERIMENTAL STUDY ON THE EFFECT OF THE DEMETHYLCHLORTETRACYCLINE ON MANDIBULAR SYMPHYSIS IN RATS (Demethylchlortetracycline이 백서 하악골결합부의 발육에 미치는 영향에 관한 실험적 연구)

  • Chang, Hyun Il
    • The korean journal of orthodontics
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    • v.10 no.1
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    • pp.37-43
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    • 1980
  • This study was undertaken to assess the effects of demethylchlortetracycline on bone growth of mandibular symphyseal region in rats. Demethylchlortetracycline at 30mg/kg body weight, respectively, were daily administered by mouth to the sewen female rats from 10th day of pregnancy to 13th day. Thirty six new-born rats from these experimental animals were used for histological examination at 1, 5, 10, 15, 20, 25 and 30 days. All these new-born rats were killed by an overdose of ether. Mandibular bodies were removed and fixed in $10\%$ neutral formalin,. Carney and aceton. Specimens were embedded, sectioned and stained with H-E, Van Gieson, PAS and prepared for alkaline phosphatase. The results were as follows; 1. Until erupting of incisors, hyaline cartilage was located in relatively large symphyseal space, but bone trabeculae of ossifying area at incisal side were arranged irregularly in experimental group. 2. During this period, PAS reaction was moderately positive, but alkaline phosphatase reaction was slightly positive. 3. By erupting or incisors, symphyseal space appeared narrower like control group, but alkaline phosphatase reaction tended to slow down. 4. By erupting of molars, symphyseal space appeared muck narrower, and cartilane plate was reduced and aealed off like control group. Alkaline phosphatase reaction tended to slow down severely.

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Cloning and Expression of Alkaline Phosphatase Gene from Schizosaccharomyces pombe

  • Kang, Sung-Won;Cho, Young-Wook;Park, Eun-Hee;Ahn, Ki-Sup;Lim, Chang-Jin
    • BMB Reports
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    • v.34 no.3
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    • pp.262-267
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    • 2001
  • A cDNA coding alkaline phosphatase (AP) homologue was isolated from a cDNA library of Schizosaccharomyces pombe by colony hybridization. The nucleotide sequence of the cloned cDNA appeared to lack the N-terminal coding region. The genomic DNA encoding alkaline phosphatase homologue was isolated from S. pombe chromosomal DNA using PCR. The amplified DNA fragment was ligated into plasmid pRS315 to generate the recombinant plasmid pSW20. The DNA insert was subcloned as two smaller fragments for nucleotide sequencing. The sequence contains 2,789 by and encodes a protein of 532 amino acids with a molecular mass of 58,666 daltons. The S. pombe cells containing plasmid pSW20 showed much higher AP activity compared with the yeast cells with vector only This indicates that the cloned AP gene apparently encodes AP The predicted amino acid sequence of the S. pombe AP shares homology with those of other known APs.

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