• 대한방사성의약품학회지
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• 제4권2호
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• pp.65-72
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• 2018

Prostate specific membrane antigen (PSMA) is a cell surface membrane protein, which is overexpressed in most prostate cancer. Recently, PET imaging with $[^{68}Ga]$PSMA-HBED-CC has been widely used for the diagnosis of recurrent prostate cancer and the studies on the diagnostic potential of $^{64}Cu$-labeled PSMA ligands reported actively. In this study, we monitored with biological evaluation in vivo and PET imaging of $^{64}Cu$-labeled PSMA ligand ($[^{64}Cu]$PSMA-617). The radiolabelling efficiency and stability of $[^{64}Cu]$PSMA-617 were confirmed by radio-thin layer chromatography. The radiolabeling efficiency of $[^{64}Cu]$PSMA-617 showed over 95%, and stabilities of intact remained over 98% in both human and mouse serum for 48 h. In normal male mice, in vivo uptake of $[^{64}Cu]$PSMA-617 in several organs was measured at 2, 4, 6, 24, 48 h after injection. Rapid blood clearance was observed for $[^{64}Cu]$PSMA-617. The high uptake was observed in the lung, liver, intestines and kidneys at 2 h postinjection, but was low in the other organs (1-2 %ID/g) at 4 h. The dynamic PET/CT images of 22RV1 tumor-bearing nude mice were acquired during 60 min and additionally acquired 24 h and 48 h after injection. In dynamic PET images, $[^{64}Cu]$PSMA-617 uptake ratio in tumors versus muscle was increased as time elaplsed until 60 minutes and remained in tumors at 48 h. In these results, the PET/CT imaging using $[^{64}Cu]$PSMA-617 in prostate cancer is expected to be useful for the diagnosis and treatment of prostate cancer patients.

• 한국발생생물학회지:발생과생식
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• 제4권2호
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• pp.161-173
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• 2000

컴퓨터 세포동결기를 이용하여 생쥐 배아를 동결할 때 액체질소 (L$N_2$)의 분사속도가 해빙 후 배아의 미세구조, 기능 및 발달에 미치는 영향을 알아보고자 하였다. 이를 위해 배아는 동결을 하지 않은 대조군 (control) 및 동결군에서 L$N_2$의 분사속도에 따라 고속분사군 (120 infusion/min group 1), 저속분사군 (50 infusion/min; group 2)으로 나누었다. ICR 계열의 생쥐의 2 세포기 배아를 사용하였으며, 동결 및 해빙은 저속동결-급속해빙 방법을 사용하였다. 각 군에 따라 해빙 후 배아의 생존율과 세포질이 양호하고 분절화가 없는 2세포기 배아를 대상으로 포배 발달율 및 할구수를 측정하였다. 공초점 현미경을 이용하여 배아 내에서의 $H_2O$$_2$, 활성 미토콘드리아의 분포, 막전위차 및 actin filament를 측정하였으며, TUNEL 방법을 이용하여 DNA 분절화를 확인하였다. 동결-해빙 후 건강한 2 세포기 배아의 회수율은 group 1 (50.7%)에 비해 group 2 (34.6%)에서 현저히 감소했다 (p<0.05). 포배기 배아의 발생율 (86.7%, 76.7% vs. 44.0%)과 할구수 (79.5$\pm$12.9, 71.6$\pm$8.0 vs. 62.5$\pm$4.7)는 대조군 혹은 group 1에 비해 group 2에서 유의한 차이를 보였다 (p<0.05). H$_2$0$_2$의 상대적 강도는 group 2에서 유의하게 증가하였다 (15.3$\pm$3.0, 16.6$\pm$1.6 vs. 23.4$\pm$1.8, p<0.05). 활성 미토콘드리아의 분포는 정상적인 배아에서는 균등하게 분포하는 반면 배발달이 정지된 배아에서는 원형질막 주위에 몰리고 응집된 양상을 보였다. 그러나 대조군, group 1, group 2에서는 모두 균등하게 분포하여 각 군간에 차이가 없었다. 미토콘드리아의 JC-1 염색 결과는 대조군과 group 1의 경우 590 nm의 파장으로 발산되는 미토콘드리아가 group 2에 비하여 유의하게 증가하였다 (17.2$\pm$3.8, 17.4$\pm$1.3 vs. 13.2$\pm$2.0, p<0.05). 2세포기 배아내 미세섬유 (actin filament)는 대조군 및 group 1의 경우 균일하게 분포하는 반면, group 2에서는 부분적인 결손과 응집현상이 관찰되었다. DNA 분절율 (30.8%, 36.0% vs. 65.6%; p<0.05)은 group 2에서 유의하게 증가하였다. 동결시 액체질소의 분사속도는 해빙 후 배아 발달에 매우 중요한 요인으로 작용하며, L$N_2$의 분사속도의 증가는 동결과 정에서 하강 온도의 미세한 변화를 감소시켜 세포내 골격구조와 미토콘드리아의 상해를 감소시켜 $H_2O$$_2$의 발생과 DNA 분절화를 감소시켜 배아 발생을 호전시키는 것으로 사료된다.

• The Korean Journal of Physiology and Pharmacology
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• 제16권4호
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• pp.255-264
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• 2012

The structures of the intact synaptosomal plasma membrane vesicles (SPMVs) isolated from bovine cerebral cortexs, and the outer and the inner monolayer separately, were evaluated with 1,6-diphenyl-1,3,5-hexatriene (DPH) and 1,3-di(1-pyrenyl)propane (Py-3-Py) as fluorescent reporters and trinitrophenyl groups as quenching agents. The methanol increased bulk rotational and lateral mobilities of SPMVs lipid bilayers. The methanol increased the rotational and lateral mobilities of the outer monolayers more than of the inner monolayers. n-(9-Anthroyloxy)stearic acid (n-AS) were used to evaluate the effect of the methanol on the rotational mobility at the 16, 12, 9, 6, and 2 position of aliphatic chains present in phospholipids of the SPMVs outer monolayers. The methanol decreased the anisotropy of the 16-(9-anthroyloxy)palmitic acid (16-AP), 12-(9-anthroyloxy)stearic acid (12-AS), 9-(9-anthroyloxy)stearic acid (9-AS), and 6-(9-anthroyloxy)stearic acid (6-AS) in the SPMVs outer monolayer but it increased the anisotropy of 2-(9-anthroyloxy)stearic acid (2-AS) in the monolayers. The magnitude of the increased rotational mobility by the methanol was in the order at the position of 16, 12, 9, and 6 of aliphatic chains in phospholipids of the outer monolayers. Furthermore, the methanol increased annular lipid fluidity and also caused membrane proteins to cluster. The important finding is that was far greater increase by methanol in annular lipid fluidity than increase in lateral and rotational mobilities by the methanol. Methanol alters the stereo or dynamics of the proteins in the lipid bilayers by combining with lipids, especially with the annular lipids. In conclusion, the present data suggest that methanol, in additions to its direct interaction with proteins, concurrently interacts with membrane lipids, fluidizing the membrane, and thus inducing conformational changes of proteins known to be intimately associated with membranes lipids.

• The Korean Journal of Physiology and Pharmacology
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• 제1권1호
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• pp.27-34
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• 1997

In the present study, it was aimed to further indentify the intracellular action mechansm of cromakalim and levcromakalim in the porcine coronary artery. In intact porcine coronary arterial strips loaded with fura-2/AM, acetylcholine caused an increase in intracellular free $Ca^{2+}$ $([Ca^{2+}]_i)$ in association with a contraction in a concentration-dependent manner. Cromakalim (1 ${\mu}M$) caused a reduction in acetylcholine-induced increased $[Ca^{2+}]_i$ not only in the mormal physiological salt solution (PSS) but also in $Ca^{2+}$-free PSS (containing 1 mM EGTA). In the skinned strips prepared by exposure of tissue to 20 .${\mu}M$ B-escin, inositol 1,4,5-trisphosphate ($IP_3$) evoked an increase in $[Ca^{2+}]_i$, but it was without effect on the intact strips. The $IP_3$-induced increase in $[Ca^{2+}]_i$ was inhibited by cromakalim by 78% and levcromakalim by 59% (1 .${\mu}M$, each). Pretreatment with glibenclamide (a blocker of ATP-sensitive $K^+$ channels, 10 .${\mu}M$) and apamin (a blocker of small conductance $Ca^{2+}$-activated $K^+$ channels, 1 .${\mu}M$) strongly blocked the effect of cromakalim and levcromakalim. However, charybdotoxin (a blocker of large conductance $Ca^{2+}$-activated $K^+$ channels, 1 .${\mu}M$) was without effect. In addition, cromakalim inhibited the $GTP{\gamma}S$ (100 .${\mu}M$, non-hydrolysable analogue of GTP)-induced increase in $[Ca^{2+}]_i$. Based on these results, it is suggested that cromakalim and levcromakalim exert a potent vasorelaxation, in part, by acting on the $K^+$ channels of the intracellular sites (e.g., sarcoplasmic reticulum membrane), thereby, resulting in decrease in release of $Ca^{2+}$ from the intracellular storage site.

• 한국환경생태학회지
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• 제30권1호
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• pp.130-134
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• 2016

$CO_2$는 기공 메커니즘에 어떤 영향을 주는가? 햇빛에 의해 유도된 기공 열림에서 독립적인 $CO_2$의 효과를 분리해서 본다는 것은 어려운 일이기 때문에, $CO_2$에 의한 기공 열림 메커니즘은 아직 명확하게 밝혀지지 않은 실정이다. 기공은 또한 $CO_2$ 농도에 따라 다르게 반응 할 수 있다. 기공 열림과 닫힘의 식물의 생체적인 리듬도 관여하므로, $CO_2$의 반응에 대한 해석은 많은 요소들을 고려해야 한다. 세포간극 내강 ($C_i$)의 감소된 $CO_2$에서는 기공을 열린다는 것이 일반적으로 정해진 사실이다. 기공 열림의 정도를 결정하는 것은 삼투 물질이고, $CO_2$가 삼투 물질의 수송에 영향을 준다고 가정하는 것이 $CO_2$가 기공 메커니즘에 영향을 주는 유일한 방법이다. 그러나 $CO_2$가 어떻게 공변세포 내의 삼투물질 농도에 영향을 주는지 그 메커니즘은 불분명하다. 지금까지, $CO_2$는 공변세포의 삼투퍼텐셜을 증가시키는 이온과 유기물이 어떻게 공변세포 막을 통한 수송 메커니즘이 이루어지는지는 알려진 것이 없다. 따라서 이 연구에서는 $CO_2$에 의한 삼투물질들의 공변세포 막 투과성에 대해 초점을 두었다. 잎을 일정한 농도의 $CO_2$에 노출할 때 $CO_2$-관련된 반응들이 나타난다. 빛에 의한 기공 열림의 가설은 $K^+$, $Cl^-$, 슈크로스 그리고 말산$^{2-}$를 포함하는 공변세포 내 삼투물질 농도의 증가에 있다. $CO_2$가 $H^+$를 세포 밖으로 방출하는 것을 나타내는 막의 과분극 (hyperpolarization)을 유도했다는 보고가 있다. 이는 $CO_2$가 막 투과성에 관련된 첫 번째 증거이다. 온전한 잎에서 $CO_2$는 빛에 의해 유도된 막의 과분극보다 3~4 배까지 공변세포의 막 과분극을 유도했다. 이러한 결과들은 $CO_2$가 막 투과성에 영향을 주는 인지질 이중층과 수송단백질의 물리적인 특성에 변화를 초래한다는 것을 의미한다.

• Molecules and Cells
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• 제24권3호
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• pp.378-387
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• 2007

The Bcl-2 family of proteins interacts at the mitochondria to regulate apoptosis. However, the anti-apoptotic Bcl-2 and $Bcl-X_L$ are not completely localized to the mitochondria. In an attempt to generate Bcl-2 and $Bcl-X_L$ chimeras that are constitutively localized to the mitochondria, we substituted their C-terminal transmembrane tail or both the C-terminal transmembrane tail and the adjacent loop with the equivalent regions from Bak or Bax mutant (BaxS184V) as these regions determine the mitochondrial localization of Bak and Bax. The effects of these substitutions on subcellular localization and their activities were assessed following expression in HeLa and CHO K1 cells. The substitution of the C-terminal tail or the C-terminal tail and the adjacent loop of Bcl-2 with the equivalent regions from Bak or the Bax mutant resulted in its association with the mitochondria. This change in subcellular localization of Bcl-2 chimeras triggered cells to undergo apoptotic-like cell death. The localization of this Bcl-2 chimera to the mitochondria may be associated with the disruption of mitochondrial membrane potential. Unlike Bcl-2, the loop structure adjacent to the C-terminal tail in $Bcl-X_L$ is crucial for its localization. To localize the $Bcl-X_L$ chimeras to the mitochondria, the loop structure next to the C-terminal tail in $Bcl-X_L$ protein must remain intact and cannot be substituted by the loop from Bax or Bak. The chimeric $Bcl-X_L$ with both its C-terminal tail and the loop structure replaced by the equivalent regions of Bak or Bax mutant localized throughout the entire cytosol. The $Bcl-X_L$ chimeras that are targeted to the mitochondria and the wild type $Bcl-X_L$ provided same protection against cell death under several death inducing conditions.

• 한국환경과학회지
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• 제6권2호
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• pp.125-131
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• 1997

The mechanism of stomatal closing in response to $O_2$ was indirectly investigated by using $H_2O_2$ which is the intermediate product of $O_2$ metabolites. Stomata in epidermal strips close in response to $H_2O_2$. The effect of $H_2O_2$ on stomatal closing was dependent on the concentration of $H_2O_2$. 10 ppm $H_2O_2$ showed a clear effect on stomatal closing and 1000 ppm $H_2O_2$ induced complete stomatal closing after the treatment of 3 hours. Stomatal closing by $H_2O_2$ in intact leaf was also observed by measuring the diffusion resistance with porometer. It was found that the stomatal closing by $H_2O_2$ was not mediated by $Ca^{2+}$, and that was a different result observed in stomatal closing by water stress. Reversely, $Ca^{2+}$ showed a great inhibition on stomatal closing. The leakage of K+ in epidermal strips was doubled in response to $H_2O_2$ when it was campared to the control. 10 ppm $H_2O_2$ decreased photosynthetic activity. Fv/Fm representing the activity of Photosystem II was reduced about 4 % in 10 ppm $H_2O_2$ and 8 % in 100 ppm $H_2O_2$ In the treatment of 1.5 hour. However, stomatal closing by 10 ppm $H_2O_2$ was reduced about 56 %. According1y, it can be suggested that stomatal closing by $H_2O_2$ is related with the decrease of photosynthetic activity, but it was chiefly induced by the change of the membrane permeability. Key words Commelina communis, stomatal closing, $H_2O_2$, $Ca^{2+}$, photosynthesis.

• Archives of Plastic Surgery
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• 제34권6호
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• pp.765-770
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• 2007

Purpose: Peritendinous adhesion is one of the most notorious complication after the flexor tendon injury. In this study, $Alloderm^{(R)}$(LifeCell Corp., Branchburg, N.J.), which is the decellularized human dermal analogue with its intact native basement membrane components, was used for the prevention of peritendinous adhesions following flexor tendon repair. Methods: Thirty New Zealand white male rabbits were divided equally into 3 groups. In all groups, the flexor digitorum profundus of the third finger of the right back foot was cut totally and repaired by modified Kessler suture technique. Following tendon repair, $Alloderm^{(R)}$ was wrapt around the repaired tendon in the first group and sodium hyaluronate gel was sprayed to the operation field in the second group. In the control group, no external material was applied. The right back foot were immobilized for 6 weeks to optimize the formation of adhesion ingrowth. After death, the third finger that repaired tendons and sheaths was removed en bloc. We checked range of motion. and studied histologically for all groups. Results: The experimental groups had better range of motion than the control group. We checked that the range of motion was 73.5 degrees in $Alloderm^{(R)}$ group, 55.9 degrees in the hyaluronic acid group, and 38.3 degrees in the control group. in the histological study, the experimental group had less adhesions compared with the control group. Conclusion: This study concludes that $Alloderm^{(R)}$ can decrease peritendinous adhesions following flexor tendon repairs in rabbits. We think the method could be used in clinical cases.

• 한국임상수의학회지
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• 제31권2호
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• pp.112-116
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• 2014

2개월령 수컷 핏불테리어종 개 (체중 1.01 kg)가 심한 복부 팽만, 운동 불내성, 성장 지연, 노란 비루와 식욕부진을 주증으로 진료 의뢰 되었다. 영상 진단상에 비정상적인 막으로 분할된 확대된 우심방, 심한 수축기 폐동맥 제트 혈류 (최고혈류 5.66 m/s) 그리고 심방 중격에서의 우좌 단락혈류가 관찰되었다. 임상증상 및 진단학적 결과에 기초하여, 본 증례는 심한 폐동맥 협착과 우좌 단락의 난원공 개존증이 합병된 우측 삼중심방증으로 진단 되었다. 비안정적인 환자의 상태 때문에 수술적 혹은 중재적 치료는 시도되지 않았다. 그 개는 수혈, 산소공급 그리고 심장 관련 약물적 치료(예, 실데나필, 스피로노락톤, 에날라프릴)로 회복되었다.

• 한국수정란이식학회지
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• 제13권2호
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• pp.179-190
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• 1998

Prolactin (PRL) surge in cycling rats at proestrous afternoon has previously been reported as an inducer of apoptotic cell death of luteal cells. This death-inducing action of PRL seeins unusual, because PRL can he categorized as a cell-survival factor, if other known physiological functions of PRL are taken into account. In this study, the apoptotic action of PRL was assessed in cultured cells prepared from rat luteal tissue and underlying molecular /cellular mechanism of PRL-induced luteolysis was analyzed. The latest crop of corpora lutea (CLs) were enucleated from rat ovaries at 18:00 h on the proestrous day before the next ovulation. Donor rats were pretreated with CB154, a dopamine agonist, in order to he exempted from the endogenous PRL surge. The harvested GLs were dispersed and cultured with or without PRL (2$\mu$g /ml) for 24 or 48 h. An addition of PRL to the culture medium changed the parameters indicative of cell death via apoptosis: a decrease in cell viability (MTT) and an increase in chromatin condensation. Most of the DNA breakdown in nuclei induced by PRL occurred in steroidogenic cells which were identified by 3$\beta$-HSD activity staining, and the number of 3$\beta$-HSD-positivecells were significantly decreased. Interestingly, most of the cells with an apoptotic nucleus adhered to one or more intact and seemingly non-steroidogenic cells. Because the expression of Fas has heen shown to be abundant in murine ovary, and Fas is known to have an exact physiological role in occurrence of apoptotic cell death, the membrane form-Fas ligand (rnFasL) was quantified in the cell lysate. An addition of PRL increased expression of mFasL. Moreover, an addition of concanavalin A (ConA), a T-cell specific activator, in place of PRL, enhanced the apoptotic parameters. Cumulatively, the apoptotic PRL action was addressed to cells unknown than steroidogenic lute~ cells. The most prohable candidate for the direct target cells is Tcells in the luteal tissue that can express mFasL in response to PRL.