• The Korean Journal of Physiology and Pharmacology
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• 제13권4호
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• pp.273-279
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• 2009

The accumulation of ${\beta}$-amyloid (A${\beta}$) aggregates is a characteristic of Alzheimer's disease (AD). Furthermore, these aggregates have neurotoxic effects on cells, and thus, molecules that inhibit A${\beta}$ aggregate formation could be valuable therapeutics for AD. It is well known that aggregation of A${\beta}$ depends on its hydrophobicity, and thus, in order to increase the hydrophilicity of A${\beta}$, we considered using citrate, an anionic surfactant with three carboxylic acid groups. We hypothesized that citrate could reduce hydrophobicity and increase hydrophilicity of A${\beta}_{1-40}$ molecules via hydrophilic/electrostatic interactions. We found that citrate significantly inhibited A${\beta}_{1-40}$ aggregation and significantly protected SH-SY5Y cell line against A${\beta}_{1-40}$ aggregates-induced neurotoxicity. In details, we examined the effects of citrate on A${\beta}_{1-40}$ aggregation and on A${\beta}_{1-40}$ aggregates-induced cytotoxicity, cell viability, and apoptosis. Th-T assays showed that citrate significantly inhibited A${\beta}_{1-40}$ aggregation in a concentration-dependent manner (Th-T intensity: from 91.3% in 0.01 mM citrate to 82.1% in 1.0 mM citrate vs. 100.0% in A${\beta}_{1-40}$ alone). In cytotoxicity and viability assays, citrate reduced the toxicity of A${\beta}_{1-40}$ in a concentration-dependent manner, in which the cytotoxicity decreased from 107.5 to 102.3% as compared with A${\beta}_{1-40}$ aggregates alone treated cells (127.3%) and the cell viability increased from 84.6 to 93.8% as compared with the A${\beta}_{1-40}$ aggregates alone treated cells (65.3%). Furthermore, Hoechst 33342 staining showed that citrate (1.0 mM) suppressed A${\beta}_{1-40}$ aggregates-induced apoptosis in the cells. This study suggests that citrate can inhibit A${\beta}_{1-40}$ aggregation and protect neurons from the apoptotic effects of A${\beta}_{1-40}$ aggregates. Accordingly, our findings suggest that citrate administration should be viewed as a novel neuroprotective strategy for AD.

• 대한한의학회지
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• 제27권2호
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• pp.122-133
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• 2006

Objectives : The amyloid $\beta$-protein ($A\beta$) is the principal component of the senile plaques characteristic of Alzheimer's disease (AD) and elicits a toxic effect on neurons in vitro and in vivo. Many environmental factors including antioxidants and proteoglycans modify $A{\beta}toxicity$. In this study, we have investigated the protective effects of water- and organic solvent-extracts of Hemerocallis fulva root fractions pre-extracted with methanol on $A\beta$-induced oxidative cell death in cultured rat pheochromocytoma (PC12) cells. Methods : For this study, we used MTT reduction assay for detection of protective effects of water- and organic solvent-extracts of Hemerocallis fulva root fractions pre-extracted with methanol on $A{\beta}_{25-35}$-induced cytotoxicity to PC12 cells. We also used cell-based $\beta$-secretase assay system to investigate the inhibitory effect of water- and organic solvent-extracts of Hemerocallis fulva root on $\beta$-secretase activity. Results : We previously reported that methanol extracts of Hemerocallis fulva root strongly attenuated cytotoxicity induced by the three $A\beta$ fragments ($A{\beta}_{25-35},\;A{\beta}_{1-42}\;A{\beta}_{1-43}$) to both SK-N-MC and PC12 cells. In the present study, we found that butanol-, ethylacetate-, chloroform-, and water-extracts of Hemerocallis fulva root fractions pre-extracted with methanol had strong protective effects against $A{\beta}_{25-35}$-induced cytotoxicity to PC12 cells and inhibitory potency to $\beta$-secretase activity. Conclusion : These results suggest that butanol-, ethylacetate-, chloroform-, and water-extracts of Hemerocallis fulva root fractions pre-extracted with methanol may contain the protective component(s) against $A\beta$-induced cell death in PC12 cells as well as inhibitory component(s) to $\beta$-secretase activity.

• 분석과학
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• 제26권1호
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• pp.106-112
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• 2013

Phosphodiesterase (PDE)는 세포내의 cAMP를 분해하는 효소로 세포의 신호 전달에 중요한 기능을 수행하는 것으로 알려져 왔다. 각각의 PDE들은 N-말단의 서열을 통해 세포 내 특정 부위로 이동되어 기능을 수행한다. 이전의 연구를 통해 바다달팽이인 군소에서 새롭게 클로닝된 ApPDE4 long-form이 원형질막과 시냅스전 뉴런의 말단에 발현됨을 확인하였다. 그러나, 현재까지 이러한 세포내 작용부위로의 이동, 즉 타겟팅(targeting)에 필요한 최소부위가 어디인지, 이러한 타겟팅이 세포에 미치는 영향은 무엇인지는 보고되지 않았다. 따라서, 본 연구에서는 이를 알아보기 위해 첫째, 원형질막으로 타겟팅에 필요한 최소부위를 알아 보고자 하였다. 이를 위해 다양한 결실돌연변이체를 제작하고, 이들의 이동과 분포를 확인한 결과, N-말단 13개의 아미노산만으로도 원형질막으로 타기팅에 충분하다는 것을 확인할 수 있었다. 또한, ApPDE4 N-말단의 20개 아미노산을 mRFP에 융합해서 만든 ApPDE4(N20)-mRFP를 HEK293T 세포에 과발현시킨 결과, 기포(bleb)가 생성되는 세포의 비정상적인 형태 변화가 관찰 되었다. 이러한 형태적 변화는 ApPDE4가 원형질막으로 타겟팅되는 것과 관련이 있었다. 대표적인 인지질의 하나인 PI4,$5P_2$에 선택적으로 결합함으로써 원형질막으로 타겟팅되는 단백질인 mRFP-$PLC{\delta}1$(PH)의 과발현도 ApPDE4(N20)-mRFP와 비슷한 세포의 형태적 변화가 유도됨을 확인할 수 있었다. ApPDE4의 N-말단은 PI4,$5P_2$와 같은 인지질과의 결합으로 원형질막으로 타겟팅될 수 있고, 형태적 변화를 유도하는 가능성을 제시한다.

• Journal of Korean Neurosurgical Society
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• 제29권10호
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• pp.1296-1302
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• 2000

Objective : Traumatic brain injury including diffuse axonal injury has been shown to result in a decrease in brainfree magnesium concentration, an endogenous inhibitor of calcium entry into neuron, that is associated with the development of neurological motor deficits. The goal of this study is to establish the therapeutic window during which the therapy with $MgSO_4$ and/or hypothermia improve damaged neurons by TUNEL stain. Method : Moderate brain injury was induced in 64 adult Sprague-Dawley rats, weighing 350 to 450gm each, by using a simple weight-drop device(Marmarou model). The animals were randomly assigned to four groups(sixteen rats each, a control group, a group treated with $MgSO_4$, a group treated with hypothermia, and a group treated with $MgSO_4$ and hypothermia) and the rats in each group were sacrificed and studied after 12 hrs, 24 hrs, 1 wk, and 2 wks after insult. In hypothermic group, these rats were subjected to hypothermia after injury, with their rectal temperatures maintained at $32^{\circ}C$ for 1 hour. After 1-hour period of hypothermia, rewarming to normothermic level was accomplished over 30-minute period. In the groups treated $MgSO_4$, hypothermia and $MgSO_4$ were subsequently treated with $MgSO_4$($750{\mu}moles/kg$) infused intra-muscularly at 30 minutes after trauma. Result : In all treated groups, a significant reduction in TUNEL positive cells was found in comparison with the control group each time(p<0.001). Between treatment groups, No differnce was seen 12hrs, 24hrs, and 1wk. However, hypothermic group treated with or without $MgSO_4$ showed more significant reduction in apoptotic cells than group treated with $MgSO_4$ 2 weeks after trauma(p<0.05). However, hypothermic group treated with $MgSO_4$ showed no significant reduction in apoptotic cells compared with hypothermic group(p>0.05). Conclusion : These findings suggest that both hypothermia and $MgSO_4$ significantly improve pathological changes. Otherwise simultaneously $MgSO_4$ and hypothermia treatment groups is failed to provide additional neuroprotection. These results may be relevant to the design of future clinical trials of therapeutic hypothermia and $MgSO_4$ for traumatic brain injury.

• 대한약리학회지
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• 제13권2호
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• pp.19-34
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• 1977

1) 마취가토(痲醉家兎) 및 묘(猫)에서 경뇌막외강(硬腦膜外腔)을 통(通)한 가압방법(加壓方法)으로 두개내압상승(頭蓋內驅上昇)과 혈압(血壓) 및 심박(心搏)과의 관계(關係)를 조사(調査)하였다. 2) 양동물(兩動物)에서 두개내압(頭盞內壓)을 상승(上昇)시켜 두개내압(頭蓋內壓)과 혈압(血壓)의 차(差)가 아주 적어지면 현저(顯著)한 혈압상승(血壓上昇)이 나타났다. 3) 양동물(兩動物)에서 두개내압(頭盞內壓)이 혈압(血壓)보다 높아지면 현저(顯著)한 혈압하강(血壓下降)과 현저(顯著)한 일시적(一時的)인 심박감소(心搏減少)가 나타났다. 4) Reserpine 처리동물(處理動物)에서는 두개내압상승(頭蓋內驅上昇)은 혈압하강(血壓下降), 심박감소(心搏減少)를 일으켰다. 5) 6-Hydroxydopamine 처리(뇌내)동물(處理(腦內)動物)에서는 두개내압상승(頭蓋內驅上昇)에 의한 혈압상승(血壓上昇)은 비처리동물(非處理動物)에 비(比)하여 약(弱)하였다. 6) Reserpine 처리동물(處理動物)의 측뇌실내(側腦室內)에 norepinephrine을 투여(投與)한 후(後)에는 두개내압상승(頭蓋內驅上昇)은 현저(顯著)한 혈압상승(血壓上昇)을 일으켰다. 7) 두개내압상승(頭蓋內驅上昇)에 의한 혈압상승(血壓上昇)은 두개내압상승(頭蓋內驅上昇)으로써 뇌내(腦內) noradrenergic neuron이 자극(刺戟)되여 norepinephrine 유리(遊離)가 증가(增加)한 결과(結果) 일어나는 것으로 추리(推理)하였다.

• The Korean Journal of Physiology and Pharmacology
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• 제21권5호
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• pp.475-485
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• 2017

The present study aimed to explore the neuroprotective effect and possible mechanisms of rhGLP-1 (7-36) against transient ischemia/reperfusion injuries induced by middle cerebral artery occlusion (MCAO) in type 2 diabetic rats. First, diabetic rats were established by a combination of a high-fat diet and low-dose streptozotocin (STZ) (30 mg/kg, intraperitoneally). Second, they were subjected to MCAO for 2 h, then treated with rhGLP-1 (7-36) (10, 20, $40{\mu}g/kg$ i.p.) at the same time of reperfusion. In the following 3 days, they were injected with rhGLP-1 (7-36) at the same dose and route for three times each day. After 72 h, hypoglycemic effects were assessed by blood glucose changes, and neuroprotective effects were evaluated by neurological deficits, infarct volume and histomorphology. Mechanisms were investigated by detecting the distribution and expression of the nuclear factor erythroid-derived factor 2 related factor 2 (Nrf2) in ischemic brain tissue, the levels of phospho-PI3 kinase (PI3K)/PI3K ratio and heme-oxygenase-1 (HO-l), as well as the activities of superoxide dismutase (SOD) and the contents of malondialdehyde (MDA). Our results showed that rhGLP-1 (7-36) significantly reduced blood glucose and infarction volume, alleviated neurological deficits, enhanced the density of surviving neurons and vascular proliferation. The nuclear positive cells ratio and expression of Nrf2, the levels of P-PI3K/PI3K ratio and HO-l increased, the activities of SOD increased and the contents of MDA decreased. The current results indicated the protective effect of rhGLP-1 (7-36) in diabetic rats following MCAO/R that may be concerned with reducing blood glucose, up-regulating expression of Nrf2/HO-1 and increasing the activities of SOD.

• 대한정형외과스포츠의학회지
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• 제4권1호
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• pp.29-35
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• 2005

목적: 전방십자인대 내에 기계적 수용체의 존재가 밝혀지면서 전방십자인대는 신경근육의 조절에 관여하는 기능이 있는 것으로 추측되어왔다. 하지만, 기계적 수용체의 존재에도 불구하고 전방십자인대의 말단 신경수용체로부터 대뇌피질까지의 구심성 체성감각신경로는 아직 명확히 밝혀지지 않았다. 강력한 신경친화성 표지자이며 신경연접을 건너 분열, 확산하는 pseudorabies virus(PRV)를 이용하여 전방십자인대의 구심성 체성감각신경로를 추적하고자 하였다. 대상 및 방법: PRV를 쥐의 전방십자인대에 주입한 후 약 6-7일간 신경엽접을 건너 확산하도록 허용한 후 각각의 쥐를 희생, 관류하였다. 대뇌와 척수를 체부로부터 분리하여 면역화학적으로 처리한 후 PRV의 존재여부를 확인하였다. 결과: PRV에 면역 활성화된 신경세포는 척수로부터 대뇌피질에 이르기까지 여러 위치에서 발견할 수 있었다. 특히, 뇌관의 망상활성계의 중뇌망상핵, 대뇌세포망상핵, 부거대세포망상핵, 거대세포망상핵에서 강한 양성표지반응을 보였다. 결론: 쥐의 전방십자인대의 신경말단은 척수, 뇌관 및 대뇌피질로 투사된다. 또한, 전방십자인대에 분포하는 신경말단으로부터의 대뇌피질로의 구심설 신경로에서 망상활성계가 중요한 역할을 담당할 것으로 추측된다.

• 한국산학기술학회논문지
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• 제21권8호
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• pp.572-578
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• 2020

주식 시장은 기업 실적 및 경기 상황뿐만 아니라 정치, 사회, 자연재해 등 예기치 못한 요소들에 영향을 받는다. 이런 요소들을 고려한 정확한 예측을 위해서 다양한 기법들이 사용된다. 최근 인공지능 기술이 화두가 되면서 이를 활용한 주가 예측 시도 또한 이루어지고 있다. 본 논문은 단순히 주식 관련 데이터뿐만 아닌, 거시 경제적 지표 등을 활용한 여러 종류의 데이터를 이용하여 주가에 영향을 미치는 요소에 관한 연구를 제안한다. KOSDAQ을 대상으로 1년 치 종가, 외국인 비율, 금리, 환율 데이터를 다양하게 조합한 후에 딥러닝의 Nonlinear AutoRegressive with eXternal input (NARX) 모델을 활용한다. 이 모델을 통해 1달 치 데이터를 생성하고 각 데이터 조합을 통해 만들어진 예측값을 RMSE를 통해 실제값과 비교, 분석한다. 또한, 은닉층에서 뉴런의 수, 지연 시간을 다양하게 설정하여 RMSE를 비교한다. 분석 결과 뉴런은 10개, 지연 시간은 2로 설정하고, 데이터는 미국, 중국, 유럽, 일본 환율의 조합을 사용할 때 RMSE 0.08을 보이며 가장 낮은 오차를 기록하였다. 본 연구는 환율이 주식에 가장 영향을 많이 미친다는 점과 종가 데이터만 사용했을 때의 RMSE 값인 0.589에서 오차를 낮췄다는 점에 의의가 있다.

• Journal of Korean Neurosurgical Society
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• 제63권5호
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• pp.579-589
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• 2020

Objective : No optimum genetic rat Huntington model both neuropathological using an adeno-associated virus (AAV-2) vector vector has been reported to date. We investigated whether direct infection of an AAV2 encoding a fragment of mutant huntingtin (AV2-82Q) into the rat striatum was useful for optimizing the Huntington rat model. Methods : We prepared ten unilateral models by injecting AAV2-82Q into the right striatum, as well as ten bilateral models. In each group, five rats were assigned to either the 2×10¹² genome copies (GC)/mL of AAV2-82Q (×1, low dose) or 2×10¹³ GC/mL of AAV2-82Q (×10, high dose) injection model. Ten unilateral and ten bilateral models injected with AAV-empty were also prepared as control groups. We performed cylinder and stepping tests 2, 4, 6, and 8 weeks after injection, tested EM48 positive mutant huntingtin aggregates. Results : The high dose of unilateral and bilateral AAV2-82Q model showed a greater decrease in performance on the stepping and cylinder tests. We also observed more prominent EM48-positive mutant huntingtin aggregates in the medium spiny neurons of the high dose of AAV2-82Q injected group. Conclusion : Based on the results from the present study, high dose of AAV2-82Q is the optimum titer for establishing a Huntington rat model. Delivery of high dose of human AAV2-82Q resulted in the manifestation of Huntington behaviors and optimum expression of the huntingtin protein in vivo.

• Journal of Korean Neurosurgical Society
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• 제44권6호
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• pp.375-381
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• 2008

Objective : The effects on neural proliferation and differentiation of neural stem cells (NSC) of basic fibroblast growth factor-2 (bFGF). insulin growth factor-I (IGF-I). brain-derived neurotrophic factor (BDNF). and nerve growth factor (NGF) were assessed. Also, following combinations of various factors were investigated : bFGF+IGF-I, bFGF+BDNF, bFGF+NGF, IGF-I+BDNF, IGF-I+NGF, and BDNF+NGF. Methods : Isolated NSC of Fisher 344 rats were cultured with individual growth factors, combinations of factors, and no growth factor (control) for 14 days. A proportion of neurons was analyzed using $\beta$-tubulin III and NeuN as neural markers. Results : Neural differentiations in the presence of individual growth factors for $\beta$-tubulin III-positive cells were : BDNF, 35.3%; IGF-I, 30.9%; bFGF, 18.1%; and NGF, 15.1%, and for NeuN-positive cells was : BDNF, 34.3%; bFGF, 32.2%; IGF-I, 26.6%; and NGF, 24.9%. However, neural differentiations in the absence of growth factor was only 2.6% for $\beta$-tubulin III and 3.1% for NeuN. For $\beta$-tubulin III-positive cells, neural differentiations were evident for the growth factor combinations as follows : bFGF+IGF-I, 73.1 %; bFGF+NGF, 65.4%; bFGF+BDNF, 58.7%; BDNF+IGF-I, 52.2%; NGF+IGF-I, 40.6%; and BDNF+NGF, 40.0%. For NeuN-positive cells : bFGF+IGF-I, 81.9%; bFGF+NGF, 63.5%; bFGF+BDNF, 62.8%; NGF+IGF-I, 62.3%; BDNF+NGF, 56.3%; and BDNF+IGF-I, 46.0%. Significant differences in neural differentiation were evident for single growth factor and combination of growth factors respectively (p<0.05). Conclusion : Combinations of growth factors have an additive effect on neural differentiation. The most prominent neural differentiation results from growth factor combinations involving bFGF and IGF-I. These findings suggest that the combination of a mitogenic action of bFGF and post-mitotic differentiation action of IGF-I synergistically affects neural proliferation and NSC differentiation.