• The Korean Journal of Nuclear Medicine
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• v.29 no.3
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• pp.332-342
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• 1995

This study was designed to evaluate the effects of various factors on the therapeutic effect of the I-131 labeled anti-carcinoembryonic antigen monoclonal antibody(anti-CEA antibody). Tetrazolium-based colorimetric assay (MTT) was used to compare in vitro cytotoxicity of 3 Korean colon cancer cell lines (SNU-C2A, SNU-C4, SNU-C5) for selection of proper 2 cell lines in this study. The changes of the size of tumor which was xenografted to nude mice (balb/c nu/nu) were compared in 4 groups (group treated I-131 labeled anti-CEA antibody, group treated with non-radiolabeled anti-CEA antibody, group treated with I-131 labeled anti-human chorionic gonadotropin monoclonal antibody (anti-hCG antibody) as nonspecific antibody, and group injected with normal saline as a control). Immunohistochemical staining and in vivo autoradiography were performed after excision of the xenografted tumor. The results were as below mentioned. The in vitro cytotoxic effect of I-131 labeled anti-CEA antibody is most prominent in SNU-C5 cell line between 3 cancer cell lines. The changes of xenografted tumor size in both SNU-C4 and SNU-5S cell tumors at the thirteenth day after injection of the antibodies were smallest in the group treated with I-131 labeled anti-CEA antibody (SNU-C4/SNU-C5; 324/342%) comparing with other groups, group treated with anti-CEA antibody (622/660%), group treated with I-131 anti-hCG antibody (538/546%), and control group(1030/724%)(P<0.02 in SNU-C4 and P<0.1 in SNU-C5 at the 13th day after injection of antibodies). On the thirteenth day after injection of the antibodies nude mice were sacreficed to count the radiouptake of tumor and to check the changes of tumor size. Correlations between radiouptake and change of tumor size were calculated in each groups and significant negative correlation was only obtained in the group treated with I-131 anti-CEA antibody (p<0.05). There were no correlations between antigenic expression of carcinoembryonic antigen and distribution of anti-CEA antibody in both SNU-C4 and SNU-C5 cell tumors on immunoperoxidase staining. On in vivo autoradiography the distributions of anti-CEA antibody were heterogeneous and the intensities of binding were various in SNU-C4 and SNU-C5 cell tumors. It is concluded that I-131 labeled tumor-specific monoclonal antibody, anti-CEA antibody is effective in suppressing the xenografted tumor growth and the effect is influenced by sensitivity of tumor cell itself to the radiolabeled antibody and other local factors instead of specificity of antibody.

• Journal of the korean academy of Pediatric Dentistry
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• v.28 no.3
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• pp.447-463
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• 2001

Dendroaspis natriuretic peptide (DNP), a fourth member of the natriuretic peptide isolated from the venom of the Dendroaspis angusticeps snake, has been reported to be present in human plasma and atrial myocardium and caused vasorelaxation and diuresis in experimental animals. However, it is uncertain whether they are present in peripheral organs other than the heart and its further physiological roles also remains to be clarified. To assess the possible physiological role of DNP in the salivary glands, I investigated the localization of DNP peptide in the rat salivary glands by immunohistochemistry and the binding sites for radiolabelled DNP in the rat salivary glands and oral mucosa using in vitro autoradiography. DNP immunoreactivity was widely distributed in the submandibular, sublingual and parotid glands, particularly in the ducts such as the intercalated and striated ducts, where atrial natriuretic peptide (ANP) was colocalized in consecutive sections, but not in acini. High density $^{125}I-DNP$ binding sites were localized in the epithelia of the tongue and hard palate, while low density binding sites for $^{125}I-DNP$ were also distributed in the submandibular, sublingual, and parotid glands. In the hard palate and tongue, the precise location of this binding was revealed on the basal and parabasal cells of the epithelia by emulsion microautoradiography. These results suggest that DNP may not only have a role in the salivary glands but also play a role in the regulation of growth in the oral epithelium, particularly in the hard palate and tongue.

• The Korean Journal of Mycology
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• v.23 no.2 s.73
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• pp.129-138
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• 1995

DNA fragment being able to restore in vitro activity of ${\beta}-1,3-glucan$ synthase was cloned by transformation of the Saccharomyces cerevisiae LP353 mutant strain with genomic library constructed in the YCp50. For the selection of transformants which showed no detectable phenotype linked to recovery of the defect in ${\beta}-1,3-glucan$ synthase activity, the colony autoradiography was succesfully applied. The restriction map of the cloned DNA fragment, which is 8.5-kb in length, was constructed. Both the YEplac195 and the YCp50 carrying the 8.5-kb fragment increased ${\beta}-1,3-glucan$ synthase activity of LP353 by two fold. Neither the YEplac195 nor the YCp50 carrying the 8.5-kb DNA fragment, however, complemented the temperature-dependent osmotic sensitivity which is another distinctive phenotype of LP353. Subcloning experiments indicated that a functional region was located in 4.8-kb BglII-KpnI fragment. The 4.8-kb fragment was also able to increase the level of ${\beta}-1,3-glucan$ content in cell wall as well as the resistance of cells to cell wall lytic enzyme, ${\beta}-1,3-glucanase$. The growth rate of the LP353 with 4.8-kb fragment was almost same as that of wild type strain in liquid medium with 1.2 M sorbitol at nonpermissive temperature. Taken these results together, the 4.8-kb fragment seemed to contain the BGS2 gene for ${\beta}-1,3-glucan$ synthase activity in yeast S. cerevisiae.

• The Korean Journal of Nuclear Medicine
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• v.26 no.2
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• pp.371-379
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• 1992

악성종양의 진단 및 치료에 있어서 특정 종양에 대한 항체를 이용하는 연구가 활발히 진행되고 있다. 단세포군항체를 이용한 방사면역신티그라피로 암의 조기 진단 및 영상화가 가능하고 나아가 방사면역치료는 암의 선택적 치료에 도움이 될 수 있다. 위암은 우리나라에서 가장 흔한 악성종양으로 방사면역신티그라피와 방사면역치료법이 새로운 방법으로 모색되고 있다. 이러한 진단 및 치료법의 성공여부를 결정하는 중요한 인자의 하나가 종양조직내에서 종양관련 항원의 농도와 분포이다. 따라서 본 연구에서는 단세포군 항체를 이용한 방사면역학적 방법의 임상 이용을 위한 기초 연구의 일환으로 in vitro quantitative autoradiography를 이용하여 종양 관련항원인 TAG-72와 CEA의 위암조직내 농도 및 분포를 측정하였다. 33예의 위암조직에서 얻은 동결절편을 $1.3\sim83.3$ nmol/liter의 $^{125}I$ 표지 항 TAG-72 단세포군 항체 B-72.3과 항 CEA 단세포군 항체 CEA-79로 반응시킨 후 이 표본들의 자가방사법 디지탈 영상을 H & E 염색과 immunoperoxidase염색 표본과 비교하였으며, 특정 단세포군 항체의 결합에 대한 컴퓨터 분석으로 조직내 항원의 농도와 분포를 측정하였다. TAG-72는 25예(75.7%)의 조직에서 검출되었으며 그 농도는 $8.4\sim525.3$ pmol/gram이었고, CEA는 32예 (96.9%)에서 검출되었으며 그 농도는 $8.8\sim592.9$ pmol/gram 이었다. CEA의 위암 조직내 발현농도는 중앙치가 101.7 pmol/gram 으로 TAG-72의 중앙치인 27.9 pmol/gram 보다 높았다. TAG-72의 조직내 분포는 41.4%에서 병변 부위의 암세포 분포와 일치하였고, CEA의 분포는 병변 부위의 80.5%에서 암세포와 일치하는 소견을 나타내었다. TAG-72의 농도는 점액성 선종(mucinous adenocarcinoma)과 점액함유 선종(mucin containing adenocarcinoma)에서 다른 선종보다 더 높았다. CEA의 농도는 위암의 병리학적 종류에 따른 유의한 차이가 없었다. 이상의 결과로 위암조직에서 TAG-72와 CEA항원이 다양하게 발현됨을 알 수 있었고 CEA는 TAG-72 보다 더 빈번하게 균일한 분포로 발현하는 것으로 나타났다.

• The Korean Journal of Zoology
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• v.38 no.4
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• pp.523-530
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• 1995

Specific affinity binding sites for atrial natriuretic peptide (ANP) were Investigated in the pig ovarian tissues by in vitro autoradiographic techniques. In the pig ovary, the highest binding sites for 12514abeiled rANP(l~28) were localized in the granulosa cell layer of the forncles. The binding sies on theca layer of the ovarian follicles were mainly localized in the external layer, but none was observed In the Internal layer. In the corpus luteum, the binding site was not observed. The specific bindings of 200 pM of l2Sl4abelled rANP(l~28) to granulosa and theca externa layers were reversed completely by excess concentration (1 ~4) of unlabelled rANP(l~28) but not by 10 ~ of unrelated peptides, human angiotensin II and arginine vasopressin. The binding was also displaced by 1 ~ of desiGIn18, Ser19, Gly2O, leu21, Gly22I ANP(4~2g) (C- ANF) as a spedfic ligand of the ANP clearance receptor. Therefore these results indicate ~hat the biological and the clearance ANP receptors exist in the theca externa and granulosa layer of the pig ovary, and suggest that the ANP receptors may be related with the regulatory lundion of the ovarian follicular development including oocyte maturation.

• The Korean Journal of Zoology
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• v.14 no.4
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• pp.175-180
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• 1971

The effects of X-irradiation on the mitotic activity, the chromosome aberration and the DNA synthetic pattern in synchronized human kidney cells treated with 5-AU were measured in the present experiment. When 5-AU was added, mitotic activity was markedly suppressed. After removal of the cells from the chemical, its activity proceeded synchronouly and reached peaks at hours 10. In 5-AU+100R groups, it was observed the X-ray caused mitotic delay, the irregularity of the time when mitotic peak appeared and the inhibiton of mitotic activity. In the control group, chromosome aerrations per cell was 0.030, whereas 0.147 in 5-AU treated group. In 5-AU+100R and 5-AU+200R groups, chromosome aberrations per cell were 0.583 and 0.669 respectively and the average chromosome aberrations per cell per R was 0.0035. 5-AU increased the frequency of labeled metaphases together with labeling intensity, and this is thought to be due to the accumulation of cells by 5-AU at S stage. On the contrary, X-ray decreased the labeling intensity and the frequency of labeled metaphases.

• Applied Microscopy
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• v.7 no.1
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• pp.21-43
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• 1977

This experiment was undertaken in order to localize the labeled dbcAMP (dibutyryl cyclic AMP) in oocytes whose development has been suppressed by cold dbcAMP for 6 or 19 hours in vitro. Mouse oocytes were obtained from the ovaries of 3-4 week old A strain female mice, by puncturing the Graafian follicles in the modified Krebs-Ringer bicarbonate salt solution under the dissecting microscope. Those oocytes which have intact germinal vesicle were cultured in the basic culture medium supplemented with 0.4% bovine serum albumin (BSA). Cultivation of the oocytes was carried out in a microtube developed by Cho (1974). The cultures were then incubated in a humidified 5% $CO_2$ incubator maintained at $37^{\circ}C$ for 6 or 19 hours (Donahue, 1968). DbcAMP was added to culture medium for a final concentration of 100ug/ml, and $^3H-dbc$ AMP (specific activity 13 Ci/mM) for a final concentration of $40{\mu}Ci/ml$ was also added to the medium. For electron microscopic autoradiography, those oocytes recovered from the culture were washed with phosphate buffer (pH 7.4), and immediately prefixed in a 2.5% glutaraldehyde overnight and postfixed for 2 hours at $4 ^{\circ}C$ in 1% osmium tetroxide in phosphate buffer with pH 7.4 (Palade, 1952). After fixation, the materials were dehydrated in graded alcohol series and embedded in Epon 812 mixture based on the standard procedures (Luft, 1961). The thin sections $600-700{\AA}$ thick were mounted on the grids of 200 meshes. The grids containing sections were coated with a nuclear emulsion Kodak NTB-3 and stored in a cold dark box (at $4^{\circ}C$) for 3 weeks. After exposure, the samples were developed with Kodak D-19 and stained with uranyl acetate and lead citrate. Routine observation was made with Hitachi HU-11E electron microsocope. The results of the observation were as followings: 1. It was found that the labeled dbcAMP penetrated the egg plasma membrane and dispersed at random in the cytoplasm. 2. It was also observed that most of the labeled dbcAMP was attached to microfibrillar lattices portion of the oocyte cytoplasm. There fore, it is presumed that the receptor of the dbcAMP is localized in the microfibrillar lattices of the oocyte. 3. It also seems that some other cell organells such as mitochondria, Golgi complex, cortical granules are not directly related to the action of the dbcAMP. 4. The labeled dbcAMP was neither observed in the membrane nor in the nucleus. Therefore, it seems that there is no relationship between the concentration of dbcAMP and the nuclear membranous permeability. 5. There was no difference in number of dbcAMP particles when oocytes were cultured for 6 hours and 19 hours. 6. However, it was observed that, in same of the oocytes suppressed in germinal vesicle by dbcAMP for 19 hours, cell organells were moved and concentrated to a small portion of the cytoplasm, and that the morphology of the organells greatly changed to an abnormal. form. Therefore, it is supposed that those oocytes were in the process of degeneration. From the above results, it is expected that dbcAMP penetrated the egg membrane and was bound to the receptor which seems to be located in the microfibrillar lattiees portion, and that this dbcAMP-receptor complex inhibited some enzyme system of the oocytes which are essential for the germinal vesicle breakdown.