• Title/Summary/Keyword: 면역조직화학법

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Immunohistochemical identification of porcine reproductive and respiratory syndrome virus antigen in the lungs of naturally infected piglets (돼지 생식기 호흡기 증후군 바이러스에 자연감염된 포유자돈의 폐장에서 면역조직화학법을 이용한 바이러스 항원의 확인)

  • Cheon, Doo-Sung;Min, Kyoungsub;Chae, Chanhee
    • Korean Journal of Veterinary Research
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    • v.37 no.2
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    • pp.417-423
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    • 1997
  • 돼지 생식기 호흡기 증후군 바이러스의 nucleocapsid와 반응을 하는 SDOW17 단크론항체를 이용하여 중성 포르말린에 고정시킨 자연감염된 포유자돈의 폐장에서 면역조직화학법을 이용하여 돼지 생식기 호흡기 증후군 바이러스 항원을 확인하였다. 서울대학교 수의과대학 병리학교실에 의뢰된 포유자돈들 중에서 병리조직학적으로 폐장에서 간질성 폐렴이 관찰된 포유자돈 7두를 임의로 선택하여 본 실험을 실시하였다. 간질성 폐렴의 병변으로 많은 수의 대식세포 침윤을 동반한 폐포벽 두께의 증가와 제II형 폐포세포의 비후가 관찰되었다. 검사한 7두 포유자돈중에서 6두에서 돼지 생식기 호흡기 증후군 바이러스에 대한 항체를 enzyme-linked immunosorbent assay에 의해 확인하였다. SDOW17 단크론항체를 이용한 면역조직화학염색과 간질성 폐렴의 대식세포에서 돼지 생식기 호흡기 증후군 바이러스의 항원을 검출하였고, 항원은 (주로)대식세포의 세포질에서만 진한 갈색의 양성반응이 관찰되었다. 이상 검사결과 돼지 생식기 호흡기 증후군 바이러스는 폐장의 간질과 폐포강에 분포되어 있는 대식세포에서 주로 증식하는 것으로 판명되었다. 본 실험에서 사용한 면역조직화학법은 돼지 생식기 호흡기 증후군 바이러스 감염여부를 바이러스 분리 또는 혈청검사 없이 진단하는데 사용할 수 있는 유용한 진단방법으로 판명되었다.

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Expression of Sodium-Iodide Symporter (NIS) in Thyroid Nodules: Comparison of RT-PCR and Immunohistochemical Staining Methods (갑상선 결절에서 Sodium Iodide Symporter (NIS)의 발현: RT-PCR방법과 면역조직화학염색법의 비교)

  • Bae, Sang-Kyun;Lee, Kang-Dae;Chang, Hee-Kyung
    • The Korean Journal of Nuclear Medicine
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    • v.38 no.6
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    • pp.511-515
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    • 2004
  • Purpose: The sodium-iodide symporter (NIS) expression is an important factor in determining the sensitivity of radioiodine therapy in well-differentiated thyroid cancers. Several previous studies for the expression of NIS in thyroid tissues show diverse results. To investigate whether there is difference between methods in determining the expression of NIS in thyroid tissues of patients with thyroid nodules, we measured the expression ot NIS using two different methods (RT-PCR and immunoshistochemical staining) and compared the results. Materials & Methods: We measured the expression of NIS by reverse transcriptase-polymerase chain reaction (RT-PCR) and also by immunohistochemical staining using anti-NIS antibody in thyroid cancers and other benign thyroid diseases. We compared the results of each method. We included 19 papillary carcinomas, 1 follicular carcinoma, 7 medullary carcinoma, 4 adenomas and 7 nodular hyperplasias. Results: By RT-PCR analysis, 10 of 19 papillary carcinomas expressed NIS, but 1 follicular cancer didn't express NIS. By immunohistochemical staining, 15 of 19 papaillary carcinomas express NIS, but 1 follicular lancer didn't express NIS. There was a significant correlation between the semiquautitative results of RT-PCR and immunohistochemical staining of NIS expression. (p<0.01) Conclusion: Our data demonstrated that the expression of NIS in thyroid cancers and other benign diseases investigated by RT-PCR and immunohistochemical staining correlated well each other. However, by immunohistochemical staining, more NIS expression was found.

Study on the Change of Catecholamine, Arginine Vasopressin and V1 Vasopressin Receptor Release in the Stressed Rat Brain

  • Kim, Tae-Gyun;Kim, Jee-Hee;Kim, Seung-Hee;Kang, Seog-Youn;Ki, Kyung-Chung;Huh, Young-Buhm;Lee, Song-Deuk
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1997.04a
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    • pp.85-85
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    • 1997
  • 스트레스가 유발된 랫드의 대뇌에서 Vasopressin-catecholamine pathway의 활성도를 알아보기 위해 면역화학염색법으로 vasopressin 호르몬의 분비와 catecholamine의 생성변화를 tyrosine hydroxylase (TH) 효소의 발현변화로 규명하고, arginine vasopressin (AVP)과 V1 vasopressin receptor의 유전자 발현변화를 in situ hybridization 방법을 이용하여 살펴보았다. 수컷 SD rat를 7시간동안 stress cage에 넣어 16$\pm$1$^{\circ}C$의 물에 수침구속 스트레스를 준 후 대조군과 함께 관류고정하여 brain을 적출하였다. Brain의 hypothalamus 부위를 중심으로하여 동결절편하여 면역조직화학 염색과 in situ hybridization을 시행하였다. TH 면역조직화학 염색에서 대뇌의 줄무늬체 부위의 꼬리조가비핵에서와 시상하부 부위의 내측등쪽시상하부와 흑색질부위에서 스트레스군이 대조군에 비해 TH 면역염색성이 증가되어 관찰되었으나 시상하부 부위의 시삭위핵, 뇌실주위핵, 뇌실옆핵에서는 두 군간의 큰 면역염색성의 차이는 보이지 않았다. AVP 면역조직화학 염색에서는 시삭위핵에 많은 수의 AVP 양성 신경세포체들이 밀집되어 있으며 뇌실옆핵에서는 스트레스군에서 AVP 면역염색성이 약간 증가되어 관찰되었으나 신경섬유의 분포양상은 비슷하였다. 중간융기에서는 모두 강한 염색성의 신경섬유들이 관찰되어 두 군간에 큰 차이는 없었다. AVP 유전자에 대한 in situ hybridization 결과 시삭위핵의 신경세포에서 AVP mRNA 양성반응을 관찰할 수 있었으나 다른 시상하부핵에서는 관찰할 수 없었으며, V1 vasopressin receptor에 대한 in situ hybridization 결과는 두 군의 대뇌에서 모두 양성반응을 관찰할 수 없었으며 V1 vasopressin receptor 유전자의 조직별 발현정도와 스트레스에 의한 발현량 조절을 관찰할 필요가 있다고 사료된다.

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Immunohistochemical Study of NSE in Small Cell Lung Cancer (SCLC) Combined with Serum Assay (소세포폐암에서 Neuron Specific Enolase의 면역조직 화학염색과 혈청농도에 관한 연구)

  • Kwak, Seung-Min;Kim, Hyung-Jung;Shin, Dong-Hwan;Jang, Joong-Hyun;Lee, Hong-Lyeol;Kim, Se-Kyu;Ahn, Chul-Min;Kim, Sung-Kyu;Lee, Won-Young;Lee, Kyi-Beom
    • Tuberculosis and Respiratory Diseases
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    • v.39 no.6
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    • pp.502-510
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    • 1992
  • Background: Neuron specific enolase (NSE) is a neuronal form of the glycolytic enzyme enolase which was first found in extracts of brain tissue, and later in a variety of APUD cells and neurons of the diffuse endocrine system. SCLC shares many APUD properties with normal neuroendocrine cells. NSE immunostaining and serum NSE measurement may be a useful marker of neuroendocrine differentiation in lung tumors and diagnosis of small cell carcinoma. Methods: NSE immunohistochemical staining was done and at the same time serum NSE levels were measured in 22 small cell lung cancer and 21 non small cell lung cancer which were confirmed histologically. Results: 1) NSE immunoreactivity was detected in 9 of the 18 (50%) small cell lung cancer, in 5 of the 16 non small cell lung cancer. 2) Whereas the mean value in non-small cell lung cancer group was $11.79{\pm}4.47\;ng/ml$, the mean level of serum NSE in small cell lung cancer increased up to $59.3{\pm}77.8\;ng/ml$. In small cell lung cancer patients, mean value of limited disease group was $20.19{\pm}12.91\;ng/ml$, while mean value of extended disease group was $91.9{\pm}94.2\;ng/ml$ showing statistically significant difference. If serum levels above 20 ng/ml were tentatively defined as positive, 16 of 22 (73%) patients with SCLC had positive serum NSE level, but only one patient with NSCLC did. There was no correlation between serum NSE level and immunoreactivity of NSE. Conclusion: These studies indicate that serum NSE measurement may be a useful marker for the diagnosis and disease extent and NSE immunostaining can be used to demonstrate the neuroendocrine components of lung tumor.

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Identification of Calretinin-immunoreactive AII Amacrine Cells in the Brazilian Opossum (Monodelphis domestica) (브라질산 주머니쥐(Monodelphis domestica) 망막 내에서의 calretinin 면역반응성을 가지는 AII 무축삭세포의 동정)

  • Jeong, Se-Jin;Jeon, Chang-Jin
    • Journal of Korean Ophthalmic Optics Society
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    • v.19 no.2
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    • pp.271-277
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    • 2014
  • Purpose: The purpose of this study was to investigate the immunoreactivity of calretinin in Brazilian opossum (Monodelphis domestica) retina. Calcium-binding protein calretinin is known to play a key role in calcium-mediated signal transduction. Methods: Experiments have been performed by standard immunocytochemical techniques on retina of the Brazilian opossum. Results: Calretinin-immunoreactivity was exhibited within the horizontal subpopulations, AII amacrine and ganglion cell subpopulations in the Brazilian opossum retina. Especially, all calretinin-immunoreactive AII amacrine cells also expressed parvalbumin. Conclusions: Similar to other mammalian retinas, calretinin-immunoreactivity was also observed within the AII amacrine cells in the Brazilian opossum retina. Thus, calretinin can be a marker of AII amacrine cells in the Brazilian opossum retina.

Purification of the Yolk Protein, and Identification of the Synthetic Site of Its Precursor in Eriocheir japonicus (Decapoda, Brachiura) (동남참게(Eriocheir japonicus)의 난황단백질 정제와 그 전구체의 합성부위 구명)

  • HAN Chang-Hee;BAE Hyun-Hwan
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.25 no.5
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    • pp.432-442
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    • 1992
  • To identify the histological site of synthesis of yolk protein precursor, vitellogenin, by immunocytochemical method in the freshwater crab Eriocheir japonicus, we purified the yolk protein, vitellin, from crude egg extracts, and prepared the anti-rabbit serum against vitellin. Then, the site of vitellogenin synthesis was demonstrated by immunotytochemical method with PAP(peroxidase-antiperoxidase) reaction using the rabbit antiserum aganist vitellin. Female specific serum protein was identified in female serum by immunoelectrophoresis and Ouchterlony's immunodiffusion test for mature male and female sera. Based on the immunoelectrophoresis and Ouchterlony's diffusion test for mature male and female sera and crude egg extracts using antiserum against vitellogenic female serum absorbed with male serum, the female specific serum protein was identified as vitellogenin, detected in female serum only. The major yolk protein, vitellin, was purified from the crude egg extracts by DEAE-cellulose ion exchange chromatography, followed by sepharose CL-4B gel filteration chromatography. The molecular weight of vitellin was estimated to be about 245,000 dalton by sepharose CL-4B gel filteration chromatography. from the results of immunological analysis for vitellin, it was found that the vitellin antiserum contained the antibody against vitellogenin. In the results of immunocytochemical reaction by PAP method with the rabbit antiserum against vitellin, the vitellogenic oocytes and the hepatopancreas of mature female showed positive PAP reaction, but not in follicle cells and previtellogenic oocytes nf ovary, muscle of female and mature male hepatopancreas. Therefore, it showed that the hepatopancreas of mature female is the site of vitellogenic synthesis.

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간장내 Superoxide Dismutase 측정법

  • 임동윤;고석태
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1993.04a
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    • pp.149-149
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    • 1993
  • 목적:동식물세포에는 superoxide(0$_{-2}$)의 불균화 반응을 촉매하는 superoxide dismutase (SOD)가 존재한다. 이 효소의 생리적 의의는 지금까지도 명확하게 되어있지 않는면이 많지만, 그 의의를 명확하게 하기위해서도 측정법의 확립이 필요하다. 방법: SOD측정 방법으로는 1) Cytochrome C method 2) Nitroblue tetrazoliun method (NBT법) 3) 면역학적 방법 4) 화학발광법 등이 있다. 실험 재료는 흰쥐, mouse, 토끼의 간을 이용하였으며, 또한, 노화 및 암세포를 이용한 방법을 이용하였다. 결과: Cytochrome C 방법을 통해서 각 장기조직 (신장, 간장, 폐)에서 SOD를 측정하였으며 SOD 활성이 낮은 암조직이나 배양세포에서는 NBT 방법이 측정방법으로 적합한 것으로 나타났으며 ,간장세포내에서의 SOD의 존재부위를 확인하는 방법으로는 면역 황금 표지방법을 사용하므로 간장 mitochondria 내에 Cu, Zn-SOD가 존재함을 알 수 있었다.

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Experession of Vasoactive Intestinal Peptide in the Hypothalamus of Fasting and Anorexia Mutant Mice (anx/anx) (절식시킨 생쥐와 식욕부진 돌연변이 생쥐의 시상하부에서 Vasoactive Intestinal Peptide의 발현)

  • 김미자;김영옥;김혜경;정주호
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.30 no.5
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    • pp.937-942
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    • 2001
  • The present study was conducted to identify the mechanism about the regulation of appetite by examining the expression patterns of vasoactive intestinal peptide in the hypothalamus of either fasted for 24 hours or anorexia mutant mouse. In order to investigate expression pattern of the vasoactive intestinal peptide, immunohisto-chemistry was employed along with reverse transcription polymerase chain reaction (RT-PCR) and dot blotting. Immunohistochemistry has shown that level of expression of vasoactive intestinal peptide and appetite-suppessing neuropeptide, was lower in the suprachiasmatic nucleus (SCN) and higher in the paraventricular nucleus (PVN) of the anorexia mutant group than in the comparable regions in the control group. This pattern was repeated in the fasting group, which also showed lower and higher levels of vasoactive intestinal peptide expression in the SCN and PVN respectively, In contrast, the vasoactive intestinal peptide mRNA level in the entire hypothalamus via RT-PCR and dot blotting was similar in the fasting and control groups, while it was significantly increased in the anorexia mutant group.

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Effect of CIMT on the Functional Improvement and BDNF Expression in Hemiplegic Rats Whose Somatomotor Area was Removed (체성운동영역이 제거된 편마비 흰쥐에서 억제 유도치료가 기능향상과 BDNF 발현에 미치는 효과)

  • Lim, Chang-Hun;Hwang, Bo-Gak
    • The Journal of the Korea Contents Association
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    • v.8 no.9
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    • pp.194-203
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    • 2008
  • CIMT(Constraint Induced Movement Therapy) is to improve the function and use of damaged upper limbs by not only confinement of unaffected limbs' exercise but also inducement of affected limbs' one. The purpose of the study is to verify the effect of CIMT by means of motor behaviour test and immunohistochemistry, using animal models. This study was analyzed using 40 male Sprague-Dawley rats as the experimental groups and 40 ones as the control groups. The rats were divided into two random groups : one group as an experimental group which was operated on under anesthesia and removed somatomotor regions with CIMT and the other as the control group without CIMT.Postural Reflex Test, Beam Walking Test, Limb Placement Test and Immunohistochemistry were run on the day 1, 3 , 7 and day 14 following surgery to each 10 rat. As a result, this study demonstrates that CIMT might be an effect method to verify the plasticity of central nervous system as motor behaviour test made all high scores (p<.05) and BDNF was high too in experimental groups.