• 핵의학기술
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• 제13권1호
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• pp.116-119
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• 2009

목적: 호르몬검사에서 채혈 후, 신속한 혈장분리의 과정은 검사결과의 신뢰성에 영향을 줄 수 있다. 혈중의 Adreno Corticotropic Hormone (ACTH)은 단백질의 분해에 의해 매우 불안정한 것으로 알려져 있다. 본 연구는 ACTH검사에서 ethylenediaminetetra-acetic acid (EDTA) tube의 온도가 환자 결과에 미치는 영향과 전혈에서 시간과 온도에 따른 ACTH의 안정성을 평가하였다. 방법: 전체 22명의 검체를 대상으로 하였으며, 냉장상태의 EDTA tube와 실온상태의 EDTA tube에 채혈한 후(n=18), 즉시 원심분리하여 두 그룹 간의 ACTH 결과를 비교하였다. 전혈상태의 EDTA 검체를 일정량을 나누어 분주하여, 실온에서 2시간(n=11), 4시간(n=11), 24시간(n=7), 냉장에서 24시간(n=7) 동안 방치한 후 원심분리하여 ACTH 농도를 측정하였다. 냉장 EDTA에 채혈 후 즉시 원심분리된 검체의 ACTH 결과와 실온에서 2시간, 4시간 방치된 검체의 ACTH결과를 비교하였고, 냉장에서 24시간 방치된 결과와 실온에서 24시간 방치된 결과를 비교하였다. 통계적인 분석은 paired t-test를 이용하였다. 결과: 실온상태의 EDTA tube에 채혈한 ACTH 결과가 냉장상태의 EDTA tube에 채혈한 결과보다 유의하게 낮게 나타났다(p=0.018). 채혈 후 즉시 원심분리된 ACTH 결과와 비교하여 실온에서 2시간 방치된 결과가 유의하게 낮았고 (p<0.001), 실온 2시간과 실온 4시간 사이에는 유의한 차가 없었다(p=0.907). 24시간 동안 방치된 결과를 보면, 실온보관과 냉장보관에서 유의한 차가 없었다(p=0.474). 결론: ACTH 검사 시에는 냉장고에 넣어둔 EDTA tube의 사용이 권장되며, 채혈 후 2시간 동안 실온에서 방치될 경우 결과가 유의하게 감소되므로, 신속한 원심분리가 요구된다.

• Toxicological Research
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• 제22권4호
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• pp.307-313
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• 2006

The development of in vitro assays has been recommended to screening and testing the potential endocrine disruptors (EDs). These assay systems focus only on identifying the estrogenic or antiestrogenic activity of EDs, whereas a few studies have been carried out to screen the thyroid hormone (TH) disruptors. The aim of this study was to evaluate a test system to detect TH disruptors using rat pituitary tumor $GH_3$ cells. The test system is based on the TH-dependent increase in growth rate. As expected, L-3,5,3-triiodothyronine ($(T_3)$ markedly induced a morphological change in $GH_3$ cells from flattened fibroblastic types to rounded or spindle-shaped types. $T_3$ stimulated $GH_3$ cell growth in a dose-dependent manner with the maximum growth-stimulating effect being observed at a concentration $1{\times}10^9M$. In addition, $T_3$ increased the release of growth hormone and prolactin into the medium of the $GH_3$ cells culture. Using this assay system, the TH-disrupting activities of bisphenol A (BPA) and its related compounds were examined. BPA, dimethy/bisphenol A (DMBPA), and TCI-EP significantly enhanced the growth of $GH_3$ cells in the range of $1{\times}10^{-5}M\;to\;1{\times}10^{-6}M$ concentrations. In conclusion, this in vitro assay system might be useful for identifying potential TH disruptors. However, this method will require further evaluation and standardization before it can be used as a broad-based screening tool.

• 대한수의학회지
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• 제36권3호
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• pp.699-707
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• 1996

This study was designed to examine the involvement of lead in function of target organ, and the protective effect of selenium in lead-treated guinea pigs for 8 weeks. The effects of exposure to 0.5% lead acetate(lead) and/or 4ppm sodium selenite(selenium) in feed on serveral aspects were evaluated by measuring thyroid stimulating hormone(TSH), triiodothyronine($T_3$), thyroxine($T_4$), serum biochemical activities, organ weights, and serum and organ lead concentrations in growing animals. The many indicators of endocrine function(TSH, $T_3$, and $T_4$ in serum), enzyme and biochemical activities(${\alpha}$-glutamyltranspeptidase, alkaline phosphatase, lactate dehydrogenase, triglyceride, creatinine, $Ca^{2+}$ in serum), and organ weights(kidney, spleen and testis) were correlated with lead exposure or showed significantly different mean values between the exposed and controls. These changes on some aspects were reversed by combination-fed of selenium, but did not statistically significant. The organ(kidney, liver, spleen, testis and brain) and serum lead concentrations of lead-fed group were clearly higher than that of controls. Selenium supplementation resulted in a significant protection against lead accumulation in liver and testis. These results suggest that lead can cause a toxic effect on several organ and that selenium seems to has a protective effect on specific reaction by lead-induced organic function toxicity.

• 대한수의학회지
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• 제27권1호
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• pp.27-34
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• 1987

To verify the direct effects of the thyroid hormone ($T_3$) on the rabbit heart, $T_3$-Tyrode solution in vitro was perfused on the normal atrial muscles and enzymatically isolated ventricular myocytes of the rabbit. All the experimental procedures were conducted at $35^{\circ}C$ and the same procedures were repeated after Ca. 120 minutes from the beginning of $T_3$-Tyrode perfusion. Compared to the state between the normal Tyrode solution and $T_3$-Tyrode solution, results were observed on the same cells by electrophysiological methods (conventional intracellular recording and whole cell patch clamping) as soon as possible. The results obtained were as follows : 1. Action potential duration (APD) on the left atrial muscle was reduced under the perfusion of $T_3$-Tyrode. 2. Absolute refractory Period was shortened by $T_3$-Tryrode perfusion. (117 msec./114 msec., 90 msec./78 msec.) 3. Maximal Ca currents ($i_{Ca}$) were decreased in single: ventricular myocytes under the $T_3$-Tyrode (2.98 nA) than under the normal Tyrode (6.65 nA) 4. On I-V relation, reversal potential was shifted to lower membrane potential and membrane potential showing maximal $i_{Ca}$was lowered from +10mV to -20mV by $T_3$ effect. 5. Above results were likely to explain that tachycardia in the hyperthyroid state was caused in part by the reduced repolarization phase and the reduced refractory period due to the decrease of the Ca current.

• 대한핵의학회지
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• 제14권2호
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• pp.45-51
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• 1980

In an attempt to evaluate the diagnostic significance of the serum thyroid hormones in various thyroid function states, the author measured serum $T_3$ uptake, serum $T_3$, serum $T_4$, serum free $T_4$ and free $T_3$ index in 27 cases of normal subjects, 11 cases of hypothyroidism, 152 cases of euthyroidism and 81 cases of hyperthyroidism by the radioimmunoassay method. The results were as follows: 1. The ranges of serum thyroid hormones in normal subjects were, serum $T_3$ uptake; $27.4{\sim}42.1%$, serum $T_3;\;93{\sim}245ng/dl$, serum $T_4;\;4.08{\sim}12.9ug/dl$ and serum free $T_4;\;0.57{\sim}1.53ng/dl(M{\pm}2S.D.)$. 2. Free $T_4$ index and serum $T_4$ show relatively high diagnostic value in euthyroidism group, and serum $T_3\;and\;T_4$ in hypothyroidism group, while serum $T_3$, free $T_4\;and\;T_4$ show relatively high diagnostic value in hyperthyroidism group. 3. There were significant correlation between free $T_4$ index and serum $T_4$(r=0.68), and between free $T_4$ index and serum free $T_4$(r=0.67) in hyperthyroidism group.

• Asian-Australasian Journal of Animal Sciences
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• 제29권4호
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• pp.487-499
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• 2016

The aim of the present study was to examine the effects of testosterone (T) and estradiol-$17{\beta}$ ($E_2$) on the production of progesterone ($P_4$) by granulosa cells, and of the $E_2$ on the production of $P_4$ and T by theca internal cells. In the first experiment, granulosa cells isolated from the largest ($F_1$) and third largest ($F_3$) preovulatory follicle were incubated for 4 h in short-term culture system, $P_4$ production by granulosa cells of both $F_1$ and $F_3$ was increased in a dose-dependent manner by ovine luteinizing hormone (oLH), but not T or $E_2$. In the second experiment, $F_1$ and $F_3$ granulosa cells cultured for 48 h in the developed monolayer culture system were recultured for an additional 48 h with increasing doses of various physiological active substances existing in the ovary, including T and $E_2$. Basal $P_4$ production for 48 h during 48 to 96 h of the cultured was about nine fold greater by $F_1$ granulosa cells than by $F_3$ granulosa cells. In substances examined oLH, chicken vasoactive intestinal polypeptide (cVIP) and T, but not $E_2$, stimulated in a dose-dependent manner $P_4$ production in both $F_1$ and $F_3$ granulosa cells. In addition, when the time course of $P_4$ production by $F_1$ granulosa cells in response to oLH, cVIP, T and $E_2$ was examined for 48 h during 48 to 96 h of culture, although $E_2$ had no effect on $P_4$ production by granulosa cells of $F_1$ during the period from 48 to 96 h of culture, $P_4$ production with oLH was found to be increased at 4 h of the culture, with a maximal 9.14 fold level at 6 h. By contrast, $P_4$ production with cVIP and T increased significantly (p<0.05) from 8 and 12 h of the culture, respectively, with maximal 6.50 fold response at 12 h and 6, 48 fold responses at 36 h. Furthermore, when $F_1$ granulosa cells were precultured with $E_2$ for various times before 4 h culture with oLH at 96 h of culture, the increase in $P_4$ production in response to oLH with a dose-related manner was only found at a pretreatment time of more than 12 h. In the third experiment, theca internal cells of $F_1$, $F_2$ and the largest third to fifth preovulatory follicles ($F_{3-5}$) were incubated for 4 h in short-term culture system with increasing doses of $E_2$. The production of $P_4$ and T by theca internal cells were increased with the addition of $E_2$ of $10^{-6}M$. These increases were greater in smaller follicles. These results indicate that, in granulosa cells of the hen, T may have a direct stimulatory action in the long term on $P_4$ production, and on $E_2$ in long-term action which may enhance the sensitivity to LH for $P_4$ production, and thus, in theca internal cells, $E_2$ in short term action may stimulate the production of $P_4$ and T.

• 한국발생생물학회지:발생과생식
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• 제6권1호
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• pp.55-66
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• 2002

배아줄기세포(embryonic stem cell, ESC)는 미분화상태로 지속적인 계대가 가능하며, 정상 핵형과 전 분화능(pluripotency)을 가져 생체내-외에서 분화 유도시 삼배엽성의 모든 세포로 분화 가능하다. ESC를 feeder 세포 없이 부유배양하면 배아체(embryoid body, EB)를 형성하고, 초기 배아 발생과 유사한 분화 양상을 갖는다. ESC의 분화 유도가 초기배아 발생처럼 생식호르몬(GTH: FSH, LH; steroids)의 영향을 받는지는 불명하다. 본 연구는 ESC가 분화과정중 생식호르몬처리에 의해 그들 수용체가 발현되는가를 알아보고자 하였다. 순계혈통 생쥐인 C57BL/6J에서 과배란 유도후 포배를 수획하고, 유사분열적으로 불활성화된 feeder 세포와 공배양하여, 계대배양 하는 중 배아줄기세포주(JHYl)를 확립하였다. JHY1의 alkaline phosphatase 활성과 SSEA-1, 3, 4 발현을 통해 ESC임을 확인하였다. Feeder 세포 없이 ESC를 계대배양 후 호르몬처리(FSH LH E$_2$, P$_4$, T)하에서 5일 동안 부유배양하여 배아체를 형성시키고, 이후 7일 동안 부착배양하여 분화를 유도하였다. GTH와 스테로이드의 수용체 발현 실험에서 ESC에 E$_2$ 처리에 의한 LHR의 발현 증가를 제외한 나머지 호르몬 처리군에서 ESC보다 낮은 생식호르몬의 수용체 발현이 관찰되었다. 생식호르몬을 농도별 수용체 발현 정도는 증감되지 않았다. 미분화 ESC 표지유전자인 Oct-4는 호르몬 처리군에서도 발현되었다. 각 배엽의 표지유전자들(영양세포, handl; 외배엽성, keratin와fgf-5; 중배엽성, enolase와 $\alpha$ -globin; 내배엽성, gata-4와 $\alpha$ -fetoprotein) 등의 발현 양상을 조사한 결과 호르몬 처리후 내배엽성 표지유전자외에는 발현 증가가 관찰되지 않았다. 즉 생식호르몬에 의해 gata-4, $\alpha$-fetoprotein의 발현이 증가되는 것으로 보아 내배엽성 계열로의 분화 유도가 이루어진 것으로 사료된다.

• 한국동물생명공학회지
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• 제35권2호
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• pp.155-162
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• 2020

Equine follicle stimulating hormone receptor (eFSHR) has a large extracellular domain and an intracellular domain containing approximately 10 phosphorylation sites within the G protein-coupled receptor. This study was conducted to analyze the function of phosphorylation sties at the eFSHR C-terminal region. We constructed a mutant of eFSHR, in which the C-terminal cytoplasmic tail was truncated at residue 641 (eFSHR-t641). This removed 10 potential phosphorylation sites from the C-terminal region of the intracellular loop. The eFSHR-wild type (eFSHR-wt) and eFSHR-t641 cDNAs were subcloned into the pCMV-ARMS1-PK2 expression vector. These plasmids were transfected into PathHunter CHO-K1 Parental cells expressing β-arrestin 2 enzyme acceptor fusion protein and analyzed for agonist-induced cAMP response. The cAMP response in cells expressing eFSHR-t641 was lower than the response in cells expressing eFSHR-wt. EC₅₀ values of eFSHR-wt and eFSHR-t641 were 1079 ng/mL and 1834 ng/mL, respectively. eFSHR-t641 was approximately 0.58-fold compared with that of eFSHR-wt. The maximal response in eFSHR-wt and eFSHR-t641 was 24.7 nM and 16.7 nM, respectively. The Rmax value of phosphorylation sites in eFSHR-t641 was also decreased to approximately 68.4% of that in eFSHR-wt. The collective data implicate that the phosphorylation sites in the eFSHR C-terminal region have a pivotal role in signal transduction in PathHunter CHO-K1 cells, and indicate that β-arrestin is involved in coupling the activated receptors to the internalization system.

• 한국보건간호학회지
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• 제17권2호
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• pp.224-237
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• 2003

Purpose : This study was designed to describe the relationship on climacteric symptom and health behavior according to hormone replacement therapy (HRT) in climacteric women. Method : Data were collected through self-reported questionaries which were constructed to include a climacteric symptom and health behaviors of climacteric women. The subjects for this study were 135 climacteric women between 45-65 years of age living in Kuri city. Among them. 65 were in the experimental group and the other 70 were in the control group. The data were analyzed by descriptive statistics, t-test, $x^2-test$, ANOVA, and Pearson Correlation Coefficient. Results : The results were as follows 1) There was no significant difference on the climacteric symptom between the group of women receiving HRT and those who did not (t=1.973, p=0.162). 2) The mean health behavior score for the group of women receiving HRT was significantly higher than those who did not(t=2.557, p=0.042). 3) The climacteric symptom and health behavior were positively correlated (r=.289, p=0.015) in women receiving HRT, while these were negatively correlated (r=-.242, p=0.043) in women without HRT. 4) Women in the HRT group showed higher climacteric symptom than the those In the control group in most items. 5) The mean score for health behavior was 2.76 in the hormone replacement therapy group and 2.35 for the control group. 6) For women in the experimental group, climacteric symptom was significantly different by the level of education (F=7.371, p=.001). 7) For women in the control group, climacteric symptom was significantly different by the number of children (F=2.873, p=.044), the level of education (F=5.616, p=.006), the number of abortion (F=3.231, p=.015), and the status of menstruation (F=4.129, p=.020). 8) For women in the experimental group, health behavior was significantly different by the level of education (F=7.351, p=.001), frequency of exercise (F=8.106, p=.000), and the status of menstruation (F=3.446, p=.032), 9) For women in the control group, health behavior was significantly different by frequency of the exercise (F=7.381, p=.001). Conclusion : This study results indicated that there was no association between the HRT and climacteric symptoms, while there was positive correlations between the HRT and health behavior. Therefore. it is assumed that HRT increases the positive health behavior of women.

• Animal Bioscience
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• 제35권3호
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• pp.399-409
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• 2022

Objective: Follicle-stimulating hormone (FSH) is the central hormone involved in mammalian reproduction, maturation at puberty, and gamete production that mediates its function by control of follicle growth and function. The present study investigated the mutations involved in the regulation of FSH receptor (FSHR) activation. Methods: We analyzed seven naturally-occurring mutations that were previously reported in human FSHR (hFSHR), in the context of equine FSHR (eFSHR); these include one constitutively activation variant, one allelic variant, and five inactivating variants. These mutations were introduced into wild-type eFSHR (eFSHR-wt) sequence to generate mutants that were designated as eFSHR-D566G, -A306T, -A189V, -N191I, -R572C, -A574V, and -R633H. Mutants were transfected into PathHunter EA-parental CHO-K1 cells expressing β-arrestin. The biological function of mutants was analyzed by quantitating cAMP accumulation in cells incubated with increasing concentrations of FSH. Results: Cells expressing eFSHR-D566G exhibited an 8.6-fold increase in basal cAMP response, as compared to that in eFSHR-wt. The allelic variation mutant eFSHR-A306T was not found to affect the basal cAMP response or half maximal effective concentration (EC₅₀) levels. On the other hand, eFSHR-D566G and eFSHR-A306T displayed a 1.5- and 1.4-fold increase in the maximal response, respectively. Signal transduction was found to be completely impaired in case of the inactivating mutants eFSHR-A189V, -R572C, and -A574V. When compared with eFSHR-wt, eFSHR-N191I displayed a 5.4-fold decrease in the EC₅₀ levels (3,910 ng/mL) and a 2.3-fold decrease in the maximal response. In contrast, cells expressing eFSHR-R633H displayed in a similar manner to that of the cells expressing the eFSHR-wt on signal transduction and maximal response. Conclusion: The activating mutant eFSHR-D566G greatly enhanced the signal transduction in response to FSH, in the absence of agonist treatment. We suggest that the state of activation of the eFSHR can modulate its basal cAMP accumulation.