• Korean Journal of Veterinary Research
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• v.44 no.1
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• pp.99-103
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• 2004

The aim of the present study was to develop a sensitive and specific assay for the diagnosis of alcelaphine herpesvirus 1 (AlHV-1) which is a cause agent of malignant catarrhal fever in ruminants. A1HV-1 is a gamma herpesvirus, which is frequent latent, and it is often difficult to detect its antigens or specific nucleic acids because of its low genomic copies in the infected tissues. In this study, polymerase chain reaction (PCR)-dot blot hybridization (DBH) assay for detecting AlHV-1 DNA was developed and evaluated for its sensitivity and specificity as comparison with PCR and DBH alone. The developed PCR-DBH was more sensitive than PCR or DBH alone and also very specific. The results showed that the sensitivity of PCR-DBH were higher and stronger than those of PCR and DBH alone. This PCR-DBH assay can be applied efficiently to confirm the presence of AlHV-1 virus on clinical samples and to differentiate specifically between AlHV-1 infection and other viral infections.

• Korean Journal of Veterinary Service
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• v.37 no.1
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• pp.29-33
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• 2014

This study aimed at finding a fast, sensitive, and efficient protocol for molecular identification of intracellular protozoa Toxoplasma (T.) gondii. For molecular detection of T. gondii, we developed a polymerase chain reaction coupled with dot blot hybridization assay (PCR/DBH). For DBH analysis, the amplified DNA of T. gondii tachyzoite was labeled by incorporation of digoxigenin. The DBH assay alone was capable of detecting down to $1{\times}10^4$ pg of T. gondii genomic DNA. The PCR alone was capable of detecting down to $1{\times}10^3$ pg of T. gondii genomic DNA, whereas the PCR/DBH assay was capable of detecting down to $1{\times}10^2$ pg of T. gondii genomic DNA, indicating that sensitivity of the PCR/DBH method was approximately 10 to 100 times higher than PCR or DBH alone. Our PCR/DBH assay will be useful for confirming the presence of T. gondii on the samples and differentiating T. gondii infection from other intracellular protozoa infections.

• Journal of Genetic Medicine
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• v.4 no.2
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• pp.179-185
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• 2007

Purpose : Catecholamines are the neuro-transmitters in the sympathetic nervous system (SNS) and are activated by stress stimulus. Tyrosine hydroxylase (TH) and Dopamine-${\beta}$-Hydroxylase (DBH) are very important enzymes in the catecholamine synthesis. Corticotropin releasing hormone (CRH) is released in the process of reacting to stresses. The aim of this study is to find out what effects immobilization stresses have on the expression of TH, BDH and CRH mRNA in a rat's brains. Methods : We compare expression levels in rat's brains of TH, DBH and CRH mRNA induced by immobilization stresses between the test group and controled group. The expression levels of TH, DBH and CRH mRNA are measured by RT-PCR and the Western Blotting Analysis (WBA). Results : In brains and adrenal glands of the immobilization stress group, the expression levels of TH and DBH mRNAs are significantly two to three times higher (P<0.01), and CRH mRNAs are approximately one and a half times higher (P<0.05) than those of controlled group. Conclusion : This study suggest that the expression levels of TH, DBH and CRH mRNAs are activated by stress stimulus in a rat's brains and adrenal glands.

• Journal of Physiology & Pathology in Korean Medicine
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• v.17 no.3
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• pp.633-642
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• 2003

A pain is the symptom which defends against noxious stimulus about a human body, it is known that if the periphery of perceptive nerve were stimulated by a physical or chemical factors, the stimulation is induced by transmission to pain center in the cerebral cortex according to pain conduction tract. The treatment of pain is to decrease a stimulus that causes a pain or block off a nerve transmitting a stimulus or puts on a way to calm down pain center, but It is for adjustment of a pain to be the most representative in acupuncture among various ways to cure a pain in Oriental medicine. However, the analgesic effect of an individual response to acupuncture stimulation shows marked individual variations, so these days genetic a few approach is attempted. On this the author determined that the responding group was appointed those whose tail flick latency (TFL) responding time delayed the minimum of 30 % comparing with basal reaction time. For those whose TFL time had shorter than 30 % was grouped as a non-responding group. And then the hypothalamus of each group was dissected and RNA was further purified. After synthesizing cDNA using oligo dT primer, products were finally applied to the PCR. The results were as follows; The ratio of responding group to non-responding group was 6:4. Ach T (acetylcholinesterase T subunit), BF-I (Brain factor-I), DBH (Dopamine β-hydroxylase) and PNM (Phosphotidylethanolamine N-Methyltransferase) were revealed significantly in the responding group. Cathepsin B and Tau were revealed significantly in the non-responding group. The PCR results show that Ach T, BF-I, DBH and PNM are expressed abundantly in the responding group, where as cathepsin B and tau are abundant in the non-responding group. These results suggest that the analgesic effect on acupuncture stimulation is related to regulation of neurotransmitter as well as neurodegeration of cerebrum.

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2002.11a
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• pp.114-114
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• 2002

포유동물에서 뇌와 부신에서 합성.분비되는 카테콜아민(Catecholamine, CA)계 신경전달물질인 dopamine(DA), norepinephrine(NE), epinephrine(E)은 체내 각종 생리현상의 조절에 필수적이며, 생식과 관련지어서는 시상하부-뇌하수체 간 GnRH-gonadotropin 호르몬 축의 활성을 조절하는 기능 외에도 번식과 관련된 여러 행동양식을 조절함이 잘 알려져 있다. 본 연구는 CA 생합성 효소들인 tyrosine hydroxylase(TH), dopamine beta-hydroxylase(DBH), phenylethanolamine-N-methyltransferase(PNMT)의 유전자 발현에 미치는 sex steroid의 영향을 조사하였다. 성숙한 암컷 횐쥐(SD strain)의 난소를 제거하고 1주 경과 후 vehicle(sesame oil; OVX+Oil 실험군) 또는 estradiol 17$\beta$(235ug/m1; OVX+E$_2$실험군)이 든 silastic capsule(길이 14mm; 내경 1.55mm; 외경 3.125mm)을 48 시간 동안 처리한 뒤 희생시켰다. 적출된 조직으로부터 RNA를 추출한 후 semi-quantitative RT-PCR을 시행하였다. (i) TH의 발현 정도는 OVX+Oil 군에서는 시상하부) substantia nigra(SNc)) 부신 순으로, OVX+E$_2$군에서는 SN글 부신) 시상하부 순으로 나타났다. TH 발현에 미치는 estradiol의 효과로 SNc과 부신에서는 유의한 증가를 보인데 비해 시상하부에서는 유의한 감소를 관찰하였다. (ii) DBH 발현 정도는 OVX+Oil군에서는 SNc> 부신> 시상 하부 순으로, OVX+E$_2$군에서는 부신＞ SNc＞ 시상하부 순이었다. DBH 발현에 미치는 estradiol의 효과로 SNc에서는 유의한 감소, 부신에서는 유의한 증가, 그리고 시상하부에서는 통계적 유의성은 없으나 감소하는 경향을 보였다. (iii) PNMT의 발현의 경우 SNc와 시상하부에서는 기보고된 바와 같이 alternative splicing에 의해 110bp 차이의 크고 작은 두 형태의 cDNA(PNMTI & PNMTs)가 증폭되었으나 부신에서는 작은 cDNA 만이 관찰되었다. PNMTs의 발현 정도는 OVX+Oil군과 OVX+E$_2$군에서 공히 부신＞ 시상하부＞ SNc 순이었고, PNMTI의 발현은 SNc가 시상하부 보다 다소 높은 경향이었으나 유의성은 없었다. PNMTs 발현에 미치는 estradiol의 효과로 SNc에서는 유의한 감소, 부신에서는 유의한 증가, 그리고 시상하부에서는 통계적 유의성은 없으나 증가하는 경향을 보였다. 본 연구에서는 CA 생합성 효소들의 유전자 발현의 조절에 미치는 estrogen 의 영향이 세포 기원이 neural crest cell인 부신 수질은 물론 뇌의 상이한 지역간에서도 조직특이적임을 관찰하였다. 이러한 결과는 각 조직에서의 estrogen 수용체 유형의 차이 혹은 작용 모드와 각 효소 유전자 발현 사이에 중요한 상관관계가 있음을 시사한다.

• Development and Reproduction
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• v.6 no.2
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• pp.117-122
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• 2002

Dopamine(DA), norepinephrine(NE), and epinephrine(E) belong to a class of neurotransmitters known as catecholamine (CA) which are synthesized and secreted by mammalian brain and adrenal medulla. CA regulate several behavior patterns connected with breeding, and regulate GnRH-gonadotropin hormone axis' vitality between hypothalamus-pituitary gland linking with reproduction freeze. The present study examined effects of sex steroid hormone on the transcriptional activities of CA biosynthesis enzymes, tyrosine hydroxylase(TH), dopamine $\beta$ -hydroxylase(DBH), and phenylethaolamine-N-methyl transferase(PNMT). Mature female rats were ovariectomized(OVX) and implanted with 17 $\beta$-estradiol(E$_2$: 500 $\mu\textrm{g}$/ml) or sesame oil. Forty-eight hours after implantation all the animals were sacrificed. Total RNAs were extracted immediately and were applied to semi-quantitative reverse transcription-polymerase chain reaction(RT-PCR). The expression level of TH was appeared by hypothalamus > SNc> adrenal medulla orders in OVX＋Oil group, and by SNc > hypothalamus) adrenal medulla orders in OVX+E$_2$ group. Treatment with E$_2$ significantly increased TH expression in SNc and adrenal medulla but in hypothalamus, the reduced TH expression was observed. The expression level of DBH was appeared by adrenal medulla > SNc > hypothalamus orders in OVX＋Oil group and in OVX＋E$_2$ group. Administration of E$_2$ significantly reduced DBH expression in SNc, and increased in adrenal medulla. Two cDNA products, large(PNMT1) and small(PMNTs) species of 110bp difference, were amplified in SNc and hypothalamus, but only PNMTs was observed in adenal medulla. The PNMTs expression level was in the order of adrenal medulla > hypothalamus > SNc in both OVX＋Oil and OVX＋E$_2$ group. The PNMTs expression in SNc and adrenal medulla was significantly increased byE$_2$. The present report demonstrated that estrogen effects on transcriptional activities for CA biosynthethic enzymes were tissue specific in adrenal medulla as well as different region of brain. These results suggest that it might be crucial relationship between the type of estrogen receptor and CA enzyme gene expression.