• The Journal of the Korean bone and joint tumor society
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• v.5 no.1
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• pp.9-16
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• 1999

A new method of limb sparing by resection, extracorporeal irradiation and reimplantation has several theoretical advantages. This method preserves the mobility of a joint and avoids the problem of loosening or breakage of tumor prosthesis. This study involved using extracorporeal irradiated autogenous joint transplantation for reconstruction after en bloc resection, and observed the periods of functional union and histological changes in irradiated tissue of the knee joint. This study also aimed to clarify whether the degeneration of articular cartilage is induced in rabbits by a single 50Gy dose of irradiation at the knee joint. Twenty New Zealand rabbits about three kilograms were randomized into two groups of 10 rabbits each. In group 1, as control, we resected the knee joint followed by reimplantation without irradiation. Group 2 received extracorporeal irradiation on the resected knee joint followed by reimplantation. Following are the results of these observations. The osteotomy site showed external callus formation in the roentgenographic finding eight weeks after reimplantation. There was marked degenerative changes in the collagen fiber of the irradiated anterior cruciate ligament and meniscus during the fourth week, but new blood-vessel formation was observed in the vicinity. There was degenerative changes in the collagen fiber of articular cartilage treated extracorporeal irradiation at four and eight weeks in the scanning electron micrographic findings. These findings was in contrast to those of subchondral bone which showed decreased cellularity and empty lacuna at four and eight weeks. Autoradiography demonstrated active [$^3H$]uridine incorporation by irradiated chondrocyte at eight weeks after reimplantation. These results indicate that when destruction of the articular cartilage and soft tissue of the knee joint is not severe, extracorporeal irradiation and reimplantation can be used with several advantage in maintaining movement of the joint while avoiding problems of tumor prosthesis and rejection, and therefore extracorporeal irradiated autogenous joint transplantation can be used as a limb-sparing procedure for temporary biological spacer in the childhood bone tumor around the knee.

• Clinical and Experimental Reproductive Medicine
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• v.21 no.1
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• pp.121-130
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• 1994

Expression of gonadotropin releasing hormone(GnRH) has been described in the rat ovary. It remains, however, unkown whether GnRH is synthesized as a prohormone. Therefore, this study was performed to verify the expression of pro-GnRH by in situ hybridization and further to investigate the effect of gonadotropin on GnRH or GnRH mRNA in rat ovary by immunohistochemical and in situ hybridization techniques. Adult female Sprague-Dawely rats were used and the estrous cycle was synchronized by intraperitoneal injection of pregnant mare's serum gonadotropin(PMSG). Ovaries were fixed with 4% paraformaldehyde and embedded with G.C.T. compound and cut by cryostat. For immunohistochemistry, avidin-biotin peroxidase complex(ABS) method was employed and for in situ hybridization, $^{35}S$-end labeled oligonucleotide was used and followed by autoradiography. By in situ hybridization using GnRH oligomer and GAP(GnRH associated protein) oligomer, GnRH mRNA and GAP mRNA were co-localized in the fullicular cells, luteal cells, interstitial cells and theca cells. GnRH or GnRH mRNA signals in the ovary increased by human chorionic gonadotropin(hCG) injection. At the 3 and 6 hrs after hCG injection, the number of GnRH and GnRH mRNA containing cells increased rapidly and the density of GnRH and GnRH mRHA culminated at 9 hrs after heG injection. With the follicular development, the high expression of GnRH and GnRH mRNA was also observed within the follicles. After ovulation, the density of GnRH or GnRH mRNA decreased in the follicles but increased in the corpus lutea.

• The Korean Journal of Pharmacology
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• v.29 no.2
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• pp.195-202
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• 1993

Oxidative modification of cellular proteins and lipids may play a role in the development of diabetic complications. Diabetic cardiomyopathy has been suggested to be caused by the intracellular $Ca^{2+}$ overload in the myocardium, which is partly due to the defect of calcium transport of the cardiac sarcoplasmic reticulum (SR). In the present study, the possible mechanism of the functional defect of cardiac SR in diabetic rats was studied. Both of the maximal $Ca^{2+}$ uptake and the affinity for $Ca^{2+}$ were decreased in the diabetic rat SR in comparison with the control. To investigate whether the functional defect of the cardiac SR in streptozotocin-induced diabetic rat is associated with the oxidative changes of cardiac SR proteins, the carbonyl group content and glycohemoglobin levels were determined. The increase in carbonyl group content of cardiac SR (2.30 nmols/mg protein, DM; 1.78, control) and in glycohemoglobin level $(13{\sim}17%,\;DM;\;3{\sim}5%,\;control)$ were observed in the diabetics. The extent of increase in calcium transport by phospholamban phosphorylation was greater in the diabetic cardiac SR membranes than that in the control. The phosphorylation levels of phospholamban, as determined by SDS-PAGE and autoradiography with $[{\gamma}^{32}P]ATP$, were increased in diabetic cardiac SR. These results suggest that the impaired cardiac SR function in diabetic rat could be a consequence of the less-phosphorylation of phospholamban in the basal state, which is partly due to the depleted norepinephrine stores in the heart. Furthermore, the oxidative damages in cardiac SR membranes might be one of the additional factors leading to the diabetic cardiomyopathy.

• 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.

• Journal of the Mineralogical Society of Korea
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• v.5 no.1
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• pp.14-21
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• 1992

Allanite crystals rich in rare-earth elements(REE) occur in soil developed on top of anorthositic rocks in the Jungsu-ri area of Okjong-myun, Hadong-run, where large Ti orebodies are embedded in the bed rock. In this study allanite is investigated mainly by transmitted light microscopy, electron microprobe analysis, atomic absoption spectrophotometry, X-ray diffraction, infrared spectrocopy. In addition, its specific gravity and micro=indentation hardness value are measured. Allanite occurs with max. dimension of $3cm{\times}6cm$ and coexists with quartz, epidote, zircon, biotite and muscovite. It shows nearly nonmetamict crystallinity, although ${\alpha}$-particles bombardment from the disintegration of the radioactive element Th is detected by an autoradiography. The allanite is particularly enriched in REE(19.88-23.99 wt.%), but is deficient in CaO(8.35-10.29wt.%). Genesis of the allanite in this area is not understood yet. It is, however, assumed to have been formed from magmatic fluid rich in REE and Ti, based on the facts that it ocexists with zircon and that it has high $TiO_2$(0.89-1.13 wt.%) whose concentration is significant in the country rocks.

• The Korean Journal of Pharmacology
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• v.29 no.1
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• pp.121-130
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• 1993

This study was done to classify the proteins involved in the specific phosphorylation using the rat peripheral blood lymphocytes (rPBL) stimulated with mitogens, phorbol 12-myristate 13-acetate (PMA) and concanavalin A (Con A). The lymphocytes were incubated with $^{32}P-orthophosphate$ before PMA or Con A stimulation. The migration patterns of the phosphorylated proteins of mitogen-treated rPBL in two dimensional electrophoretic fields were analyzed after autoradiography. The stimulation of the lymphocytes with PMA and Con A increased the phosphorylation of thirteen protein fractions. The phosphorylation intensities of the protein spots differ to the treatments of the cells with specific kinase inhibitors, H-7 and W-7. These protein fractions were grouped into 3 classes, namely, PKC-mediated, CaM kinase-mediated, and other kinase mediated proteins. The effect of the duration of the stimulation on the phosphorylated behaviors occurred concurrently, not sequentially, although each individual protein fraction had a different time for the peak phosphorylation during the stimulation period upto 30 minutes. The phosphoproteins found in the cytosolic soluble fraction were phosphorylated prior to those in the pellet, whose phosphorylations were sustained at a high level for over 10 minutes. The above results suggest that the early events in lymphocyte activation involve 3 different sets of proteins which are phosphorylated by CaM kinase, PKC and other kinases, and those kinases do not work sequentially, but rather, independently or cooperatively.

• Journal of Aquaculture
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• v.2 no.1
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• pp.9-20
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• 1989

This study was taken to isolate transferrin fractions from the sera of tilapia(Oreochromis niloticus) by physico-chemical analyses such as the rivanol precipitation, iron-staining method, SDS-polyacrylamide gel electrophoresis and $^{59}FeCl_3$ autoradiography, and to calculate gene frequencies by using Hardy-Weinberg Law. The results obtained in this experiment were summarized as follow : 1. The transferrin fraction is composed of several components possessing relative lower electrophoretic mobilities and higher molecular sizes than the albumin components. 2. When different staining method was compared with transferrin in band, it was not found difference. 3. It was concluded that the optimun ratio of rivanol to serum was 2 : 1 and this ratio was used in all further fractionation. 4. The molecular weight of transferrin component was about 70,000 $\pm$ 2,000. 5. Tilapia transferrin fractionations were found to be polymorphic. 6. There transferrin variants(A, B and C) have been found in tilapia(Oreochromis niloticus) and Tf types were assumed to be controlled by three codominant alleles Tf A, Tf B and Tf C. Six different phenotypes can be theoretically expected Tf AA, Tf AB, Tf AC, Tf BB, Tf BC, and Tf CC. Only five types of these were observed and Tf CC types(homozygotes) was not found. 7. The frequencies of the three allele Tf A, Tf B and Tf C were 0.795, 0.15 and 0.055 respectively.

• 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.