• 대한치과교정학회지
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• 제24권1호
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• pp.115-123
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• 1994

To study bone resorption mechanism, effect of LPS on the $^{45}Ca$ release from fetal rat ulnae and radii, and effects of carbonic anhydrase inhibitors on the LPS-induced bone resorption in organ culture were studied. Ulnae and radii were removed from 19 day old fetal rats, prelabelled by subcutaneous injection of $200{\mu}Ci\;^{45}CaCl_2$ into their mother on the 17th day of gestation. Radioactivities of $^{45}Ca$ released into media were determined after 24, 48 and 72 hours. Effects of LPS and carbonic anhydrase inhibitors were observed by the ratio of $\%$ release of $^{45}Ca$ between paired control and experimental group. The observed results were as follows : 1. $LPS(1{\mu}g/ml)$ supplemented in media for 72hours increased the $^{45}Ca$ release significantly after 48 and 72 hours of culture and $LPS(10{\mu}g/m1)$ increased the $^{45}Ca$ release significantly after 72 hours of culture. 2. LPS-induced $^{45}Ca$ release was not inhibited significantly by 1mM sulfanilamide but inhibited significantly by 10mM sulfanilamide after 48 and 72 hours of culture. 3. LPS-induced $^{45}Ca$ release was not inhibited significantly by 0.1mM dichlorphenamide but inhibited significantly by 1mM dichlorphenamide after 48 and 72 hours of culture. 4. LPS-induced $^{45}Ca$ release was not inhibited significantly by 1mM acetazolamide but inhibited sighificantly by 5mM acetazolamide after 72 hours of culture.

• The Korean Journal of Physiology and Pharmacology
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• 제4권2호
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• pp.113-120
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• 2000

To investigate the mechanism of smooth muscle contraction induced by emptying of intracellular $Ca^{2+}$ stores, we measured isometric contraction and $^{45}Ca^{2+}$ influx. $CaCl_2$ increased $Ca^{2+}$ store emptying- induced contraction in dose-dependent manner, but phospholipase C activity was not affected by the $Ca^{2+}$ store emptying-induced contraction. The contraction was inhibited by voltage-dependent $Ca^{2+}$ channel antagonists dose dependently, but not by TMB-8 (intracellular $Ca^{2+}$ release blocker). Both PKC inhibitors (H-7 and staurosporine) and tyrosine kinase inhibitors (genistein and methyl 2,5-dihydroxycinnamic acid) significantly inhibited the contraction, but calmodulin antagonists (W-7 and trifluoperazine) had no inhibitory effect on the contraction. The combined inhibitory effects of protein kinase inhibitors, H-7 and genistein, together with verapamil were greater than that of each one alone. In $Ca^{2+}$ store-emptied condition, $^{45}Ca^{2+}$ influx was significantly inhibited by verapamil, H-7 or genistein but not by trifluoperazine. However combined inhibitory effects of protein kinase inhibitors, H-7 and genistein, together with verapamil were not observed. Therefore, this kinase pathway may modulate the sensitivity of contractile protein. These results suggest that contraction induced by emptying of intracellular $Ca^{2+}$ stores was mediated by influx of extracellular $Ca^{2+}$ through voltage-dependent $Ca^{2+}$ channel, also protein kinase C and/or tyrosine kinase pathway modulates the $Ca^{2+}$ sensitivity of contractile protein.

• 한국토양비료학회지
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• 제20권2호
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• pp.153-159
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• 1987

천연(天然) 제오라이트, 벤토나이트, 버미큐라이트에 의한 $^{42}K$, $^{45}Ca$ 흡착(吸着)이 Frendlich와 Langmuir 등온흡착방식(等溫吸着方式)에 맞는지, 또한 이들 흡착체(吸着體)에 흡착(吸着)된 방사성(放射性) 핵종(核種)이 $NH_4$에 의해서 탈착(脫着)되는 정도(程度)를 이들 광물(鑛物)의 결정구조적(結晶構造的) 특성(特性)과 관련시키면서 상호(相互) 비교(比較)해 봤다. 우선 $^{42}K$, $^{45}Ca$ 흡착(吸着)은 두 가지 흡착방식(吸着方式)에 잘 맞는다. Langmuir 최대흡착량(最大吸着量)은 버미큐라이트를 제외(除外)하고는 제오라이트와 벤토나이트 모두 $^{42}K$ 흡착량(吸着量)이 $^{45}Ca$ 흡착량(吸着量) 보다 많다. $^{42}K$ 흡착량(吸着量)의 크기를 보면 zeolite(Zt) > Bentonite (Bt) > Vermiculite (Vt)의 순(順)이고 $^{45}Ca$의 경우는 Bt>Vt>Zt이다. 제오라이트가 $^{42}K$를 $^{45}Ca$ 보다 더 많이 흡착(吸着)하는 것은 이온반경(半徑)과 제오라이트 공동(空洞)의 크기와 밀접한 관계가 있는 것으로 보여진다. Ca 이온반경(半徑)은 작기 때문에 공동(空洞)으로 부터 빠져 나온다. 벤토나이트가 $^{45}Ca$를 더 흡착(吸着)하는 것은 하전밀도(荷電密度)가 K의 하전밀도(荷電密度) 보다 크기 때문이라고 본다. $^{42}K$, $^{45}Ca$의 흡착강도(吸着强度)는 Langmuir 정수(定數)(k)와 비교(比較)해 보면 $^{42}K$의 경우는 k값이 $^{45}Ca$의 k값 보다 작다. k값이 작으면 흡착강도(吸着强度)는 약(弱)하다고 본다. $^{42}K$의 탈착율(脫着率)은 Zt>Bt>Vt 의 순(順)이고 $^{45}Ca$는 Vt>Zt>Bt 의 순(順)이다. 버미큐라이트의 $K^+$에 대한 선택적 고정은 제외하고 k값이 작은 것이 탈착율(脫着率)은 높다. 결론적(結論的)으로 제오라이트는 다른 개량재(改良材)보다 $^{42}K$를 더 흡착(吸着)하고 방출(放出)도 많다. 벤토나이트는 다른 개량재(改良材)보다 $^{45}Ca$를 더 흡착(吸着)하나 탈착(脫着)은 적다.

• The Korean Journal of Physiology
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• 제29권2호
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• pp.269-277
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• 1995

Membrane vesicles were prepared by differential centrifugation from epithelial cells of porcine trachea. Total activity of microsomal ATPases was measured spectrophotometrically by a coupled enzyme assay. The steady-state activity of the enzyme was $329{\pm}10$ nmol/min mg protein. Thapsigargin, a specific antagonist of intracellular $Ca^{2+}-ATPase$, inhibited about 50% of the activity, leaving $178{\pm}18\;nmol/min .mg$ protein (n=6), indicating that the $Ca^{2+}-ATPase$ is one of the major microsomal ATPases. The microsomes used in this study appeared to be tight-sealed vesicles since they showed saturation in $^{45}Ca^{2+}$ uptake experiments. Inositol 1,4,5-trisphosphate $InsP_{3}, 4\;{\mu}M$, an agonist of $InsP_{3}$-sensitive $Ca^{2+}$ release channel ($InsP_{3}$, receptor), and Ca-ionophore A23187 $(10\;{\mu}M)$ induced $^{45}Ca^{2+}$ releases of 20% and 50% of stored $^{45}Ca^{2+}$, respectively. The addition of $(10\;{\mu}M\;InsP_{3}$ also increased the microsomal ATPase activity from $282{\pm}8$ nmol/min mg protein to $334{\pm}21$ nmol/min . mg protein in the intact vesicles. Similar increase in the activity was observed by making microsomes leaky (uncoupling) using the Ca-ionophore A23187. ;$InsP_{3}-induced$ effects were blocked by either thapsigargin or heparin suggesting that: 1) the $InsP_{3}-induced$ increase in ATPase activity is mediated by microsomal $Ca^{2+}-ATPase$, and 2) dissipation of $Ca^{2+}$ gradient across the microsomal membrane is responsible for the $InsP_{3}-induced$ effect. In order to test the dependence of the $Ca^{2+}-ATPase$ activity on the activity of $InsP_{3}-induced$ the activity of ATPases was monitored in various concentrations of free $Ca^{2+}$ using $EGTA-Ca^{2+}$ buffers. The $Ca^{2+}$-dependent biphasic change is the well-known character of $InsP_{3} receptor but not of microsomal $Ca^{2+}-ATPase$ in non-excitable cells; however, the activity of microsomal ATPase appeared biphasic and a maxim진 activity of $397{\pm}36nmol/min\;.mg$ protein was obtained in the solution containing 100 nM free $Ca^{2+}$. Below or above this concentration, the activity of ATPases was lower. These results strongly support a positive correlation of microsomal $Ca^{2+}-ATPase$ to the $InsP_{3}$ receptors in epithelial microsomes.

• Archives of Pharmacal Research
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• 제6권1호
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• pp.69-73
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• 1983

It was reported from our laboratory that the rate of deterioration of the force of contraction was slower in heart from Panax ginseng extract treated rats. Present investigation was designed to elucidate the mechanism of the slow deterioration of contractility of ginseng treated hearts. Therefore, $^{45}Ca^{2+}$ Uptake by sarcoplasmic reticulum (SR) isolated from ginseng treated rate and control rats was studied. Rate weighing 150-250g were administered orally with ginseng ethanol extract (100mg/kg) for 10 days. Cardiac SR was isolated by differential centrifugation and $^{45}Ca^{2+}$ uptake was assessed by the Millipore method. Freshly isolated SR from treated as well as control animals did not show any differences, but after incubation for 30 and 60 min at 37.deg.C, $^{45}Ca^{2+}$ uptake of control animal SR was found to be greatly depressed. The SR of treated animal possessed a greater degree of resistance to incubation. Thus it can be concluded that ginseng may have an ability to sustain the normal function of the heart by sustaining Ca accumulation by SR involved with the excitationcontraction coupling processes.

• 한국생물물리학회:학술대회논문집
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• 한국생물물리학회 1996년도 정기총회 및 학술발표회
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• pp.39-39
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• 1996

Extracts were prepared from six different poisonous mushrooms in order to identify biologically active natural ligands. The effects of these extracts were examined on the microsomal $^{45}$ Ca$^{2+}$ uptake and $^{45}$ Ca$^{2+}$ release. Five out of six species apparently inhibited the activity of total microsomal ATPases prepared from the epithelial cells of pig airway. (omitted)ted)

• 대한치과교정학회지
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• 제22권1호
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• pp.273-283
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• 1992

To find out the differences between periodontal ligament cells (PDL cells) and gingival fibroblast cells (GFB cells), alkaline phosphatase, a marker enzyme for osteoblast, was used to measure the activities and $^{45}CaCl_2$ isotope was used to find out cellular and release of $^{45}Ca$, a requisite for bone formation,. PDL cells and GFB cells from 1 to 5 passages were also measured in alkaline phosphatase activity assay. By the use of above methods, followings were concluded that the PDL cells and the GFB cells have characteristics that are different from each other. In that PDL cells showed large amount of calcium uptake and large amount of calcium release in initial stage, they seem to possess characteristics which are similar to osteoblast-like cells. 1. The PDL cells, in contrast to the gingival fibroblast, showed exceedingly high alkaline phosphatase activity which was highest at the second passage, decreasing thereon. But gingival fibroblasts cells showed no distinct differences in alkaline phosphatase activity as the passage were elapsed. 2. For both PDL cells and GF cells, the $^{45}Ca$ uptake was greatest at 2 hours period. The PDL cells showed higher measuring than GFB cells through out the whole time period. 3. Whereas the GFB cells showed slow increase of $^{45}Ca$ release as time relapsed, the PDL cells showed rapid increase of $^{45}Ca$ release.

• The Korean Journal of Physiology
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• 제28권1호
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• pp.27-35
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• 1994

We investigated the alterations in basal tone of aortic strips by changing the Ca concentration, basal $^{45}Ca$ uptake and $^3H-nitrendipine$ binding of the single cells of aortic smooth muscles in the spontaneously hypertensive rats (SHR) and Wistar-Kyoto (WKY) rats. While the basal tone of the aortic strips in WKY rats was not affected by alteration of Ca concentration, that in SHR was decreased by the removal of Ca from the bath solution and was recovered by the restoration of Ca to normal levels. This contraction increased in a Ca concentration-dependent manner and reached a maximum at 2 mM Ca. The basal tone of aorta in SHR was suppressed by verapamil $(10^{-6}M)$. The basal tone of aorta in SHR increased about 50% in the strips of endothelial rubbing, compared with that of intact endothelium. Basal $^{45}Ca$ uptake in the aortic single smooth muscle cells of SHR was greater than that of WKY (p<0.01), Specific bindings of $[^3H]nitrendipine$ in the aortic single smooth muscles of SHR and WKY were saturable. The dissociation constant $(K_d)\;was\;0.71{\pm}0.15\;and\;1.18{\pm}0.08nM$ SHR, respectively, and the difference in $K_d$ between two strains was statistically significant (p<0.03). The maximal binding capacity $(B_{max})\;was\;34.6{\pm}3.2\;and\;47.4{\pm}4.3\;fmol/10^6$ SHR respectively, and the difference of $(B_{max})$ between two strains was statistically significant (p<0.05). from the above results, it is suggested that the increase of Ca influx via potential-operated Ca channels and the increase of the number of dihydropyridine-sensitive Ca channels contribute to high basal tone of the aortic strips in SHR.

• 대한약리학회지
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• 제32권1호
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• pp.13-21
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• 1996

세포내 칼슘농도는 신경세포의 다양한 기능에 매우 중요한 역할을 하고 있다. 본 연구에서는 일차배양한 마우스 소뇌과립세포에서 니코틴성 아세틸콜린 수용체가 특정 발생단계에 발현되고 세포내 칼슘의 농도조절에 관여하는 것을 관찰하였다. 니코틴에 의한 세포내 칼슘농도의 변화는 $^{45}Ca^{2+}$나 fura-2를 사용하여 형광법으로 측정하였다. 니코틴은 마우스 소뇌과립세포내 칼슘의 농도를 최대한 증가시키는 것으로 보인다. 반면에 일차배양한 Glia 세포들에서는 $^{45}Ca^{2+}$ 농도를 증가시키지 않았다. 세포내 칼슘농도에 미치는 니코틴의 효과는 NMDA 수용체에 대한 길항제에 의하여 억제되었다. 또한 Glutamate pyruvate transminase (GPT)를 사용하여 배양액의 글루타민산을 제거하면 니코틴효과가 소실되는 것이 관찰되었다. 이러한 결과는 니코틴에 의한 세포내 칼슘농도의 변화가 세포에서 유리된 글루타민산에 의한 간접적인 효과임을 암시한다. Fura-2를 사용한 형광법으로 실험한 결과 니코틴은 two phase로 세포내 칼슘농도를 증가시키는 것을 보여주었다. NMDA 수용체 길항제와 GPT는 단지 후기 plateau상만 억제하였다. 따라서 본 연구결과는 니코틴이 직접 니코틴성 아세틸콜린 수용체를 자극하여 일시적으로 세포내 칼슘농도를 증가시키고 글루타민산을 유리하여 NMDA 수용체를 활성화시킴으로써 세포내 칼슘농도를 지속적으로 증가시키는 것으로 보여진다. 이러한 결과는 니코틴성 아세틸콜린 수용체가 특정한 발생과정에 발현되어 세포내 칼슘농도 조절에 관여함으로써 신경발생과정에서 중요한 역할을 할 수 있음을 보여주고 있다. state를 나타내는 것을 알 수 있다. 또한 $[^3H]DPCPX$를 이용한 competitive binding assay에서 0.1 mM GTP는 효현제인 PIA의 apparent affinity를 감소시켰으며, DPCPX의 apparent affinity는 증가시키고, CGS-15943에는 아무런 영향을 미치지 않았다. 이것은 상기의 $[^{35}S]GTP_{\gamma}S$ binding의 결과를 뒤받침해 주는 결과라고 생각된다.요한 역할을 할 수 있으리라 사료된다.X>$Ca^{2+}$에 의하여 활성화되는 $K^+$ 통로를 개방시킴으로 세포내 $Ca^{2+}$을 감소시켜 뇌 기저동맥의 이완반응을 매개하는 것으로 사료된다. 함량을 조정하므로, 흉선세포의 apoptosis에 억제적으로 작용할 수 있음을 시사하는 것으로 사료된다. 영양액에 의하여는 회복됨을 볼 수 있었으며 $Mg^{++}$ 증가 영양액에서는 억제, TTX 동시 투여시에는 완전히 소실되었다. 이상의 실험결과로 흰쥐 해마에서 $A_1-adenosine$ 수용체를 통한 adenosine의 NE 유리 감소는 TEA 및 4AP에 예민한 $K^+$-통로가 관여하고 여기에는 세포외액의 Ca^{++}의 농도가 중요한 인자의 하나로 관여 하는 것으로 사료된다. 영상의 질을 크게 향상 시켜 줌으로 비가역 3구획모델에서의 PGA방법을 대체할 새로운 파라메터 영상구성방법으로 적합할 것이다.관계되며, YH439는 중금속으로 유도된 조직독성에 방어효과가 있음을 지지한다.총 아미노산의 순은

• 대한약리학회지
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• 제26권2호
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• pp.127-133
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• 1990

세포막을 투과하는 cyclic AMP의 유도체인 Dibutyryl-cyclic AMP(db-cAMP)와 ad-enylate cyclase를 활성화시킴으로써 세포내에 CAMP를 증가시키는 Forskolin을 이용하여 토끼 대동맥평활근 이완작용의 기전을 검토하여 다음과 같은 결과를 얻었다. 1. Db-cAMP는 $1{\mu}M$ norepinephrine에 의한 지속성 수축을 농도의존적으로 억제하였으나 고농도의 K에 의한 수축은 억제하지 못하였다. 2. Forskolin은 $1{\mu}M$ norepinephrine에 의한 지속성 수축을 농도의존적으로 억제하였으며, 고통도의 K에 의한 수축보다 더 효과적으로 억제하였다. 3. Db-cAMP는 $1{\mu}M$ norepinephrine에 의한 $^{45}Ca$ 유입증가를 억제하였다. 4. Forskolin은 $1{\mu}M$ norepinephrine에 의한 $^{45}Ca$ 유입증가를 억제하였으며, 고농도의 K에 의한 $^{45}Ca$ 유입증가도 억제하였으나 유의차는 없었다. 5. Db-cAMP는 칼슘이온 제거용액에서 $l{\mu}M$ norepinephrine에 의한 일과성 수축을 농도의존적으로 억제하였다. 이상의 결과에서 cAMP는 수용체작동성 칼슘채널(ROCs)을 통한 칼슘이온의 유입을 억제함으로써 norepinephrine에 의한 수축을 억제하며, 고농도의 K수축 억제가 전위의존성칼슘채널(VGCs)을 통한 칼슘이온의 유입의 억제에 의한 것인지는 확실치 않다. 또한 cAMP는 norepinephrine에 의한 세포내 칼슘이온의 유리에 의한 일과성 수축도 억제한다.