• 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.

• Nutrition Research and Practice
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• 제14권3호
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• pp.203-217
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• 2020

BACKGROUND/OBJECTIVE: Centella asiatica, also known as Gotu kola, is a tropical medicinal plant native to Madagascar, Southeast Asia, and South Africa. It is well known to have biological activities, including wound healing, anti-inflammatory, antidiabetic, cytotoxic, and antioxidant effects. The purpose of this study was to determine the efficacy of extracts of C. asiatica against age-related eye degeneration and to examine their physiological activities. MATERIALS/METHODS: To determine the effects of CA-HE50 (C. asiatica 50% EtOH extract) on retinal pigment cells, we assessed the cytotoxicity of CoCl₂ and oxidized-A2E in ARPE-19 cells and observed the protective effects of CA-HE50 against N-methyl-N-nitrosourea (MNU)-induced retinal damage in C57BL/6 mice. In particular, we measured factors related to apoptosis and anti-oxidation and the protein levels of rhodopsin/opsin. We also measured glucose uptake to characterize glucose metabolism, a major factor in cell protection. RESULTS: Induction of cytotoxicity with CoCl₂ and oxidized-A2E inhibited decreases in the viability of ARPE-19 cells when CA-HE50 was administered, and promoted glucose uptake under normal conditions (P < 0.05). In addition, CA-HE50 inhibited degeneration/apoptosis of the retina in the context of MNU-induced toxicity (P < 0.05). In particular, CA-HE50 at 200 mg/kg inhibited the cleavage of pro-caspase-3 and pro-poly (ADP-ribose)-polymerase and maintained the expressions of nuclear factor erythroid 2-related factor 2 and heme oxygenase-1 similar to normal control levels. Rhodopsin/opsin expression was maintained at a higher level than in normal controls. CONCLUSION: A series of experiments confirmed that CA-HE50 was effective for inhibiting or preventing age-related eye damage/degeneration. Based on these results, we believe it is worthwhile to develop drugs or functional foods related to age-related eye degeneration using CA-HE50.

• Healthcare Informatics Research
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• 제24권4호
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• pp.335-345
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• 2018

Objectives: The retinal nerve fiber layer (RNFL) is a site of glaucomatous optic neuropathy whose early changes need to be detected because glaucoma is one of the most common causes of blindness. This paper proposes an automated RNFL detection method based on the texture feature by forming a co-occurrence matrix and a backpropagation neural network as the classifier. Methods: We propose two texture features, namely, correlation and autocorrelation based on a co-occurrence matrix. Those features are selected by using a correlation feature selection method. Then the backpropagation neural network is applied as the classifier to implement RNFL detection in a retinal fundus image. Results: We used 40 retinal fundus images as testing data and 160 sub-images (80 showing a normal RNFL and 80 showing RNFL loss) as training data to evaluate the performance of our proposed method. Overall, this work achieved an accuracy of 94.52%. Conclusions: Our results demonstrated that the proposed method achieved a high accuracy, which indicates good performance.

• 한국가시화정보학회:학술대회논문집
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• 한국가시화정보학회 2002년도 마이크로/바이오 가시화기술부문 학술강연회
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• pp.51-78
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• 2002

A number of research groups worldwide are studying electronic implants that can be mounted on retinal optic nerve/visual cortex to restore vision of patients suffering from retinal degeneration. The implants consist of a neural interface made of biocompatible materials, one or more integrated circuits for stimuli generation, a camera, an image processor, and a telemetric channel. The realization of these classes of neural prosthetic devices is largely due to the explosive development of micro- and nano-electronics technologies in the late $20^{th}$ century and biotechnologies more recently. Animal experiments showed promise and some human experiments are in progress to indicate that recognition of images can be obtained and improved over time. We, at NBS-ERC of SNU, have started our own retinal implant project in 2000. We have selected polyimide as the biomaterial for an epi-retinal stimulator. In-vitro and in-vivo biocompatibility studies have been performed on the electrode arrays. We have obtained good affinity to retinal pigment epithelial cells and no harmful effect. The implant also showed very good stability and safety in rabbit eye for 12 weeks. We have also demonstrated that through proper stimulation of inner retina, meaning vision can be obtained.

• Molecules and Cells
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• 제46권11호
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• pp.675-687
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• 2023

Accumulation of pathogenic amyloid-β disrupts the tight junction of retinal pigment epithelium (RPE), one of its senescence-like structural alterations. In the clearance of amyloid-β, the autophagy-lysosome pathway plays the crucial role. In this context, mammalian target of rapamycin (mTOR) inhibits the process of autophagy and lysosomal degradation, acting as a potential therapeutic target for age-associated disorders. However, efficacy of targeting mTOR to treat age-related macular degeneration remains largely elusive. Here, we validated the therapeutic efficacy of the mTOR inhibitors, Torin and PP242, in clearing amyloid-β by inducing the autophagy-lysosome pathway in a mouse model with pathogenic amyloid-β with tight junction disruption of RPE, which is evident in dry age-related macular degeneration. High concentration of amyloid-β oligomers induced autophagy-lysosome pathway impairment accompanied by the accumulation of p62 and decreased lysosomal activity in RPE cells. However, Torin and PP242 treatment restored the lysosomal activity via activation of LAMP2 and facilitated the clearance of amyloid-β in vitro and in vivo. Furthermore, clearance of amyloid-β by Torin and PP242 ameliorated the tight junction disruption of RPE in vivo. Overall, our findings suggest mTOR inhibition as a new therapeutic strategy for the restoration of tight junctions in age-related macular degeneration.

• Clinical and Experimental Reproductive Medicine
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• 제37권3호
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• pp.199-206
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• 2010

망막 질환에서의 줄기세포 치료는 이전까지 치료가 불가능하다고 여겨졌던 환자들에서 시력을 향상시킬 수 있는 가능성 때문에 주목 받고 있다. 본문에서는 망막 전구세포의 분화를 위해 사용되는 태아 줄기세포, 배아줄기세포 및 성체줄기세포 등 다양한 세포 종류와, 내인적, 외인적 인자 및 이식 방법에 대해 논의하였다. 망막색소상피세포뿐만 아니라 시각세포 전구체로 성공적으로 분화시킨 실험적 연구가 보고되고 있다. 줄기세포기반치료는 아직 한계가 있지만 망막 질환 환자에서 시력을 회복하기 위한 보다 근본적인 치료 방법으로 기대되고 있다.

• 생약학회지
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• 제51권2호
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• pp.131-138
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• 2020

Ultraviolet (UV)-induced damage plays a major role in ocular diseases, such as cataracts and retinal degeneration. UV irradiation can generate free radicals including reactive oxygen species (ROS), which are known to cause lipid peroxidation of cellular membranes. It has also been shown that UV can damage DNA directly and induce apoptosis. Rhynchosia volubilis Loureiro (the small black bean or yak-kong, RV) and Beta bulgaris (beet, BB) are used as health supplements. In this study, we explored the protective effects of RV and BB against UVA-induced damage in human pigment epithelial (ARPE-19) cells and in mice. RV and BB mixture and their effective constituents (cyanidin, delphidin, petunidin glycosides) improved cell viability and suppressed intracelluar ROS generation. Phosphorylation of p38 mitogen-activated protein kinase (MAPK), c-Jun N-terminal kinase (JNK), Erk1/2 was analyzed by immunoblotting. RV and BB mixture inhibited UVA-induced phosphorylation of p38 MAPK, JNK, Erk1/2 in APRE-19 cells. RV and BB treatment also showed protective effects on ocular damage in UVA-irradiated mice by increasing the levels of endogenous antioxidants such as superoxide dismutase and glutathione. RV and BB have the potential to be used in a range of ocular diseases and conditions, based on in vitro and in vivo study.

• 대한의용생체공학회:의공학회지
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• 제30권4호
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• pp.347-354
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• 2009

For the reliable transmission of meaningful visual information using prosthetic electrical stimulation, it is required to develop an effective stimulation strategy for the generation of electrical pulse trains based on input visual information. The characteristics of neuronal activities of retinal ganglion cells (RGCs) evoked by electrical stimulation should be understood for this purpose. In this study, for the development of an optimal stimulation strategy for visual prosthesis, we analyzed the neuronal responses of RGCs in rd1 mouse, photoreceptor-degenerated retina of animal model of retinal diseases (retinitis pigmentosa). Based on the in-vitro model of epiretinal prosthesis which consists of planar multielectrode array (MEA) and retinal patch, we recorded and analyzed multiunit RGC activities evoked by amplitude-modulated electrical pulse trains. Two modes of responses were observed. Short-latency responses occurring at 3 ms after the stimulation were estimated to be from direct stimulation of RGCs. Long-latency responses were also observed mainly at 2 - 100 ms after stimulation and showed rhythmic firing with same frequency as the oscillatory background field potential. The long-latency responses could be modulated by pulse amplitude and duration. From the results, we expect that optimal stimulation conditions such as pulse amplitude and pulse duration can be determined for the successful transmission of visual information by electrical stimulation.

• Molecules and Cells
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• 제45권11호
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• pp.846-854
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• 2022

Neurons make long-distance connections via their axons, and the accuracy and stability of these connections are crucial for brain function. Research using various animal models showed that the molecular and cellular mechanisms underlying the assembly and maintenance of neuronal circuitry are highly conserved in vertebrates. Therefore, to gain a deeper understanding of brain development and maintenance, an efficient vertebrate model is required, where the axons of a defined neuronal cell type can be genetically manipulated and selectively visualized in vivo. Placental mammals pose an experimental challenge, as time-consuming breeding of genetically modified animals is required due to their in utero development. Xenopus laevis, the most commonly used amphibian model, offers comparative advantages, since their embryos ex utero during which embryological manipulations can be performed. However, the tetraploidy of the X. laevis genome makes them not ideal for genetic studies. Here, we use Xenopus tropicalis, a diploid amphibian species, to visualize axonal pathfinding and degeneration of a single central nervous system neuronal cell type, the retinal ganglion cell (RGC). First, we show that RGC axons follow the developmental trajectory previously described in X. laevis with a slightly different timeline. Second, we demonstrate that co-electroporation of DNA and/or oligonucleotides enables the visualization of gene function-altered RGC axons in an intact brain. Finally, using this method, we show that the axon-autonomous, Sarm1-dependent axon destruction program operates in X. tropicalis. Taken together, the present study demonstrates that the visual system of X. tropicalis is a highly efficient model to identify new molecular mechanisms underlying axon guidance and survival.

• 한국임상수의학회지
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• 제35권4호
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• pp.126-129
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• 2018

This study was performed to describe the outcome of pars plana retinopexy with perfluoro-n-octane (PFO)-silicone oil (SiO) exchange in dogs with rhegmatogenous retinal detachment in Seoul National University Veterinary Medical Teaching Hospital (SNU VMTH) from 2014 to 2017. Nine eyes of 8 dogs were included in this study. Medical records including signalment, history, duration from onset of blindness to surgical intervention, pre-operative findings, duration from surgery to regaining vision, and post-operative complications were evaluated. No eyes were visual before surgery. Duration from onset of blindness to surgical intervention was 2-30 days (median 8 days); duration from surgery to regain vision was 1-14 days (median 6 days); follow-up time was 15-1088 days (median 69 days). Post-operative complications were divided as temporary vs permanent conditions. Temporary complications were corneal ulcer, uveitis, retinal haemorrhage, glaucoma, subconjunctival leakage of SiO, and vitreal haemorrhage. Permanent complications were anterior chamber migration of SiO, retinal degeneration, corneal degeneration, re-detachment, and cataract. Six of 9 eyes regained functional vision, five of which remained visual throughout the follow-up time while the other one lost vision after 3 months because of uveitic glaucoma. In conclusion, pars plana retinopexy with PFO-SiO exchange provided fair outcome in 66.7% cases described in this study.