• Bulletin of the Korean Chemical Society
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• 제28권12호
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• pp.2315-2318
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• 2007

Epinephrine was determined using a lab-made chemiluminescence (CL) system with air pump. Luminolsodium IO4? chemiluminescence system was employed to produce the luminescence of epinephrine. In the reaction, epinephrine was oxidized to produce superoxide or singlet oxygen by periodate in alkaline solution, which enhanced CL of luminol. For optimization, various buffers, such as phosphate, borate, and tris, were studied in this experiment. Compared to NaOH, the phosphate and borate buffer showed better reproducibility with similar sensitivity. Small amount of sample, 22 μL, was required for a measurement. The limit of quantification for epinephrine was obtained to be ~10?9 g/mL after optimization.

• 분석과학
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• 제12권2호
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• pp.99-104
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• 1999

자외선검출법과 이온크로마토그래피를 이용하여 소금물 중에 함유하는 몇 가지 음이온의 분리정량에 대하여 검토하였다. 분리관은 음이온교환수지(Dionex, AS7)를 사용하였고 용리액으로서는 sodium chloride/sodium phosphate 완충용액을 사용하였다. 단계적용리법을 적용하므로서 최적분리가 가능하였으며 8가지 음이온(iodate, bromate, nitrite, bromide, nitrate, chromate, iodide와 thiocyanate)이 40분내에 분리되었다. 여러 가지 농도의 NaCl용액(0.0 M-1.0 M)속에 존재하는 음이온들의 거동을 살펴보았다. NaCl의 농도가 진할수록 bromate, nitrite, bromide와 nitrate의 피크의 모양이 점점 넓어졌으나 chromate, iodide와 thiocyanate는 1.0 M NaCl 용액의 농도까지 피크모양이 거의 변하지 않았다. 또한 여러 가지 농도의 NaCl용액에서 음이온들의 검량곡선은 좋은 직선성을 보여주었으며, $50{\mu}L$ 시료용액에서 검출한계는 $10-720{\mu}g/L$ 이었다. 바닷물중의 bromide, nitrate와 iodide의 측정에 이방법을 적용하였다.

• Nuclear Engineering and Technology
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• 제3권3호
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• pp.141-147
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• 1971

Tetrachloro-p-tetraiodo-R-Fluorescein 및 sodium orthoiodohippurate등 요오드화 합물과 요오드화합물이 아닌 human serum Albumin을 요오드 표지함에 있어서 표지속도와 수율등을 비교 검토하였다. 위의 각각의 화합물들을 여러가지 다른 조건하에 있는 $Na^{131}$ I, 즉, 환원제를 함유한것, 환원제를 함유하지 않으며 거의 순수하게 131 I$^{-}$형으로 있는것, 및 환원제를 함유하지 않으며 상당량이 131 IO$_3$$^{-}$형으로 있는것 등을 써서 표지 반응시켰다. 환원제인 $Na_2$S$_2$O$_3$를 함유한 $Na^{131}$ I를 쓰는 경우는 위의 세가지 화합물들의 표지수율이 10% 내외로 매우 저조하였으며 환원제는 없지만 상당량이 $Na^{131}$ I$^{-}$로 존재하는 $Na^{131}$ I를 쓰는 경우도 50~60%의 낮은 수율이었다. 환원제가 없으며 대부분 $Na^{131}$ I$^{-}$로 존재하는 $Na^{131}$ I를 쓰는 경우에만 70~90%의 높은 표지수율을 보였다. 이들 반응에 관여하는 중간체에 대해서도 토의하였다.

• Clinical and Experimental Reproductive Medicine
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• 제38권4호
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• pp.216-221
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• 2011

Objective: To differentiate the human embryonic stem cells (hESCs) into the retinal pigment epithelium (RPE) in the defined culture condition and determine its therapeutic potential for the treatment of retinal degenerative diseases. Methods: The embryoid bodies were formed from hESCs and attached on the matrigel coated culture dishes. The neural structures consisting neural precursors were selected and expanded to form rosette structures. The mechanically isolated neural rosettes were differentiated into pigmented cells in the media comprised of N2 and B27. Expression profiles of markers related to RPE development were analyzed by reverse transcription-polymerase chain reaction and immunostaining. Dissociated putative RPE cells ($10^5$ cells/5 ${\mu}L$) were transplanted into the subretinal space of rat retinal degeneration model induced by intravenous sodium iodate injection. Animals were sacrificed at 1, 2, and 4 weeks after transplantation, and immnohistochemistry study was performed to verify the survival of the transplanted cells. Results: The putative RPE cells derived from hESC showed characteristics of the human RPE cells morphologically and expressed molecular markers and associated with RPE fate. Grafted RPE cells were found to survive in the subretinal space up to 4 weeks after transplantation, and the expression of RPE markers was confirmed with immunohistochemistry. Conclusion: Transplanted RPE cells derived from hESC in the defined culture condition successfully survived and migrated within subretinal space of rat retinal degeneration model. These results support the feasibility of the hESC derived RPE cells for cell-based therapies for retinal degenerative disease.

• Molecules and Cells
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• 제43권7호
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• pp.632-644
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• 2020

The molecular mechanism underlying autophagy impairment in the retinal pigment epithelium (RPE) in dry age-related macular degeneration (AMD) is not yet clear. Based on the causative role of poly(ADP-ribose) polymerase 1 (PARP1) in RPE necrosis, this study examined whether PARP1 is involved in the autophagy impairment observed during dry AMD pathogenesis. We found that autophagy was downregulated following H₂O₂-induced PARP1 activation in ARPE-19 cells and olaparib, PARP1 inhibitor, preserved the autophagy process upon H₂O₂ exposure in ARPE-19 cells. These findings imply that PARP1 participates in the autophagy impairment upon oxidative stress in ARPE-19 cells. Furthermore, PARP1 inhibited autolysosome formation but did not affect autophagosome formation in H₂O₂-exposed ARPE-19 cells, demonstrating that PARP1 is responsible for impairment of late-stage autophagy in particular. Because PARP1 consumes NAD⁺ while exerting its catalytic activity, we investigated whether PARP1 impedes autophagy mediated by sirtuin1 (SIRT1), which uses NAD⁺ as its cofactor. A NAD⁺ precursor restored autophagy and protected mitochondria in ARPE-19 cells by preserving SIRT1 activity upon H₂O₂. Moreover, olaparib failed to restore autophagy in SIRT1-depleted ARPE-19 cells, indicating that PARP1 inhibits autophagy through SIRT1 inhibition. Next, we further examined whether PARP1-induced autophagy impairment occurs in the retinas of dry AMD model mice. Histological analyses revealed that olaparib treatment protected mouse retinas against sodium iodate (SI) insult, but not in retinas cotreated with SI and wortmannin, an autophagy inhibitor. Collectively, our data demonstrate that PARP1-dependent inhibition of SIRT1 activity impedes autophagic survival of RPE cells, leading to retinal degeneration during dry AMD pathogenesis.