Pivotal role of the RanBP9-cofilin pathway in Aβ-induced apoptosis and neurodegeneration

Woo, J A,Jung, A R,Lakshmana, M K,Bedrossian, A,Lim, Y,Bu, J H,Park, S A,Koo, E H,Mook-Jung, I,Kang, D E Macmillan Publishers Limited 2012 CELL DEATH AND DIFFERENTIATION Vol.19 No.9

Neurodegeneration associated with amyloid β (Aβ) peptide accumulation, synaptic loss, neuroinflammation, tauopathy, and memory impairments encompass the pathophysiological features of Alzheimer's disease (AD). We previously reported that the scaffolding protein RanBP9, which is overall increased in brains of AD patients, simultaneously promotes Aβ generation and focal adhesion disruption by accelerating the endocytosis of amyloid precursor protein (APP) and β1-integrin, respectively. Here, we show that RanBP9 protein levels are increased by fourfold in FAD mutant APP transgenic mice. Accordingly, RanBP9 transgenic mice demonstrate significantly increased synapse loss, neurodegeneration, gliosis, and spatial memory deficits. RanBP9 overexpression promotes apoptosis and potentiates Aβ-induced neurotoxicity independent of its capacity to promote Aβ generation. Conversely, RanBP9 reduction by siRNA or gene dosage mitigates Aβ-induced neurotoxicity. Importantly, RanBP9 activates/dephosphorylates cofilin, a key regulator of actin dynamics and mitochondria-mediated apoptosis, and siRNA knockdown of cofilin abolishes both Aβ and RanBP9-induced apoptosis. These findings implicate the RanBP9–cofilin pathway as critical therapeutic targets not only for stemming Aβ generation but also antagonizing Aβ-induced neurotoxicity.

Cytological analysis of pregnancy-associated plasma protein-A expression in porcine neonatal testis

Ji-youn Kim,Keon Bong Oh,Sung June Byun,Sun-A Ock,Hwi-Cheul Lee,황성수,SangHyun Park,Wootae Ha,Jae-Seok Woo,Hyuk Song 한국수정란이식학회 2018 한국동물생명공학회지 Vol.33 No.3

The identification of biomarkers of a living tissues is essentially required to understand specific functions of the cells. In previous study, we reported IGFBP 3 as one of the putative biomarkers, by showing specific expression at porcine spermatogonial stem cells (SSCs) of early stage of porcine testis. In this study, we analyzed the expression of seven members of IGFBP family (IGFBPs) in SSCs and histological expression pattern of pregnancy-associated plasma protein-A (PAPP-A), which plays a role on the growth promoting enzyme by cleavage of IGFBPs in testis of 5 days old pig. RT-PCR analysis showed that IGFBP 1, 2, 3, 4, and 6 were expressed at high level specifically in porcine SSCs compared with whole testis. We performed immunohisotochemical staining of testis sections with PAPP-A and protein gene product 9.5 (PGP9.5) which are the known biomarkers for SSCs. We were not able to find co-expression of PAPP-A and PGP9.5; PAPP-A was expressed only in Sertoli cells and PGP9.5 expression was confirmed in spermatogonium. Additionally, we were able to confirm the GATA4 expression in Sertoli and Leydig cells as a regulator of Sertoli cell function was not detected PGP9.5 expressing cells, indicating indirect evidence of that cytolocalization of PAPP-A expression is limited in Sertoli cells. These results suggested that the PAPP-A expressed in Sertoli cells may play role on regulation of development and differentiation of testicular cells through the IGF axis in neonatal porcine testis.

청가시덩굴 추출물의 기능성 원료 표준화를 위한 지표성분 Resveratrol, trans-Scirpusin A의 분석법 개발 및 검증

권진관(Jin Gwan Kwon),정연우(Yeon Woo Jung),최윤혁(Yun-Hyeok Choi),이지은(Ji Eun Lee),정원식(Wonsik Jeong),이정아(Jung A Lee),최춘환(Chun Whan Choi),안은경(Eun-Kyung Ahn),최용문(Yongmun Choi),홍성수(Seong Su Hong) 한국식품영양과학회 2022 한국식품영양과학회지 Vol.51 No.11

본 연구는 HPLC를 이용하여 청가시덩굴 추출물을 개별인정형 건강기능식품의 기능성 원료로 개발하기 위한 원료 표준화의 일환으로, 청가시덩굴 추출물의 지표성분을 resveratrol과 trans-scirpusin A로 설정하고 이에 대한 HPLC 분석법을 확립하여 유효성의 검증을 실시하였다. 분석법 유효성 검증은 특이성, 직선성, 정확도, 정밀도, 검출한계 및 정량한계 등을 통해 분석법의 신뢰성을 검증하였으며, 그 결과 표준용액과 청가시덩굴 추출물 간의 HPLC 크로마토그램 및 UV spectrum의 일치 여부 등의 비교를 통해 다른 물질과 간섭 없이 피크가 분리된 것으로 특이성을 확인하였다. 또한 표준용액 검량선의 상관계수(R²)는 0.9999로 매우 우수한 직선성으로 관찰되어 분석에 적합한 것으로 확인되었으며, 검량선의 기울기 및 표준편차를 이용한 검출한계는 resveratrol이 0.98 μg/mL, trans-scirpusin A는 0.49 μg/mL였고 정량한계는 resveratrol이 2.98 μg/mL, trans-scirpusin A는 1.48 μg/mL로 각각 확인되었다. 청가시덩굴 추출물에 표준물질을 3개 농도 첨가하고 분석한 회수율은 resveratrol이 98.77~99.24%, trans-scirpusin A는 98.45~99.45%로 나타나 정확성이 있는 것을 확인할 수 있었다. 청가시덩굴 추출물의 조제 농도 2.2, 4.4 및 6.6 mg/mL에서 반복성은 resveratrol이 0.99~1.22%, trans-scirpusin A는 1.12~1.32%를, 실험실 내 정밀성에서는 일내 정밀성은 resveratrol이 0.67~0.87%, trans-scirpusin A는 1.18~1.33%로 나타났고 일간 정밀성은 resveratrol이 0.93~1.22%, trans-scirpusin A는 1.33~2.27%로 확인되어 본 분석법은 정밀성이 있음을 확인할 수 있었다. 이상의 분석결과를 통해 확립된 청가시덩굴 추출물의 지표성분인 resveratrol과 trans-scirpusin A의 HPLC 분석법은 적합한 시험법으로 검증되었으며, 본 시험법은 향후 청가시덩굴 추출물의 건강기능식품 기능성 원료 개발과 표준화를 위한 기초자료로 활용될 것으로 사료된다. This study was undertaken to establish an analytical method using high-performance liquid chromatography (HPLC). HPLC for the standard determination of resveratrol and trans-scirpusin A as functional ingredients in Smilax sieboldii extract. We evaluated the specificity, linearity, accuracy, precision, limit of detection (LOD), and limit of quantitation (LOQ) of various analytical methods for detecting resveratrol and trans-scirpusin A using HPLC. The specificity was confirmed by the chromatogram obtained using the HPLC analytical method. Also, the results of UV and the coefficient of correlation (R²) obtained was 0.999, which confirmed that this was a suitable analysis with high linearity. The LOD was 0.98, 0.49 μg/mL, and LOQ was 2.98, 1.48 μg/mL, which was confirmed as a suitable limit level for the analysis of resveratrol and trans-scirpusin A content in the S. sieboldii extract. The recovery of resveratrol and trans-scirpusin A content was determined to be 98.77±0.73∼99.24±1.47% and 98.45±1.18∼99.45±1.66%, respectively, indicating high accuracy. The intra-day repeatability and the intra-laboratory precision of the daily repetition were confirmed to be 0.67∼0.87%, 1.18∼1.33% and 0.93∼1.22%, 1.33∼2.27%, respectively, for trans-scirpusin A, for the relative standard deviation. These results indicate that the reported HPLC method is simple, reliable, and reproducible for the detection of resveratrol and trans-scirpusin A in S. sieboldii extract.

Synthesis of Novel Apio Carbocyclic Nucleoside Analogues as Selective A_(3) Adenosine Receptor Agonists

Hwang, Ki-Jun,Chun, Moon-Woo,Jacobson, Kenneth A.,Jeong, Lak-Shin,Lee, Jeong-A,Moon, Hyung-Ryong,Kim, Hea-Ok,Kim, Kyung-Ran,Lee, Kang-Man,Kim, Bum-Tae 이화여자대학교 약학연구소 2005 藥學硏究論文集 Vol.- No.16

On the basis of the biological activity of neplanocin A and apio-dideoxyadenosine (apio-ddA), novel apio-neplanocin A analogues 5a- d, combining the properties of two nucleosides, were stereoselectively synthesized. The apio moiety of the target nucleosides 5a-d was stereoselectively introduced by treating· lactol 10 with 37% formaldehyde in the presence of potassium carbonate. The carbasugar moiety of neplanocin A was successively built by exposing diene 12 on a Grubbs catalyst in methylene chloride. The final nucleosides 5a-d were synthesized from the condensation of the glycosyl donor 14 with nucleic bases under the standard Mitsunobu conditions. Similarly, apio-aristeromycin 6 and (N)-apio-methanocarbaadenosine 7 were derived from the common intermediate 13 using catalytic hydrogenation and Simmons-Smith cyclopropanation as key steps. All of the final nucleosides 5a-4, 6, and 7 did not show significant inhibitory activity against S-adenosylllolllocysteine hydrolase (SAH) up to 100 ㎛, maybe due to the absence of the secondary hydroxyl group at tile C3'-position, which should be oxidized by cofactor-bound NAD^(+). However, aplo-neplanocin A (5a) showed potent and highly selective binding affinity (K_(i), = 628 ± 69 nM) at the A_(3) adenosine receptor without any binding affinity at the A_(1) and A_(2A) adenosine receptors. In conclusion, we have first developed novel carbocyclic nucleosides with unnatural apio-carbasugarsusing stereoselective hydroxymethylation and RCM reaction and also discovered a new template of human A_(3) adenosine receptor agonist, which play a great rote in developing new A_(3) adenosine receptor agonist as well as in identifying the binding site of the receptor.

소화기 ; 대전 지역에서 급성 A형 간염의 유전자형에 따른 임상 특성 고찰

이영우 ( Young Woo Lee ),양현웅 ( Hyeon Woong Yang ),이진아 ( Jin A Lee ),윤기호 ( Ki Ho Yun ),양성은 ( Seong Eun Yang ),이민지 ( Min Ji Lee ),박세영 ( Se Young Park ),김새희 ( Sae Hee Kim ),이향이 ( Hyang Ie Lee ),이윤정 ( Yun Ju 대한내과학회 2011 대한내과학회지 Vol.80 No.5

목적: B형 간염이나 C형 간염 등은 유전자형에 따른 차이가 알려져 있지만, 국내에서 증가하는 IIIA 유전자형과 기존의 IA 유전자형에 의한 간염의 차이가 거의 알려진 바 없다. 따라서 대전 지역에서 발생한 급성 A형 간염의 유전자형에 따른 차이를 알고자 한다. 방법: 2009년 11월부터 2010년 6월까지 급성 A형 간염으로 진단되면서 유전자형이 확인된 81명의 환자를 대상으로 임상적 소견들과 검사실 소견들을 전향적으로 연구하였다. 결과: 평균 나이는 32.6±7.4세이며, 평균 입원 기간은 7.7±2.4일이었다. 직업은 서비스직, 사무직 그리고 교육직이 가장 많았다. 유전자형 간의 급성 간염에 따른 증상들의 종류는 별다른 차이가 없었다. 유전자형에 따른 증상의 빈도는 IIIA 유전자형에서 구토와 설사가, IA 유전자형에서는 복통과 피부 반점이 상대적으로 많았다. 검사실 소견에서는 IIIA 유전자형에서 헤모글로빈의 평균, CRP 평균, AST 최대 수치 평균은 상대적으로 유의하게 높았다. 그리고 AST와 ALT 및 총 빌리루빈 최대 수치들과 INR의 수치가 IIIA 유전자형에서 상대적으로 높게 분포하는 경향을 보였다. 결론: 대전 지역에서 유행하는 급성 A형 간염에서 IIIA 유전자형에 따른 급성 간염은 기존의 IA 유전자형과 차이가 있을 것으로 보이며, 다기관 연구를 통한 심도 깊은 연구가 필요하겠다. Background/Aims: Acute viral hepatitis A is a major health problem in Korea and the influx of genotype IIIA is thought to be one reason. We examined the differences in the clinical characteristics and laboratory findings of genotypes IA and IIIA in Daejeon. Methods: From November 2009 to June 2010, 81 patients positive for IgM anti-HAV were enrolled prospectively. The hepatitis A was genotyped using real-time polymerase chain reaction. The clinical characteristics and laboratory results were compared on the Results: The mean patient age was 32.6±7.4 years. The mean hospitalization was 7.7±2.4 days. The patient occupation varied. Clinically, vomiting and diarrhea were relatively more prevalent in genotype IIIA than in IA. Abdominal pain and skin spots were relatively more prevalent in genotype IA than in IIIA. The hemoglobin, peak aspartate aminotransferase (AST) level, and C-reactive protein were statistically higher in genotype IIIA than in IA. The distributions of the peak AST, alanine aminotransferase (ALT) and total bilirubin values tended to be perched in genotype IIIA than in IA. The international normalized ratio (INR) tended to be slightly prolonged in genotype IIIA than in IA. Conclusions: Recently, genotype IIIA of acute viral hepatitis A has become prevalent in Daejeon. Hepatitis A genotype IIIA probably causes worse laboratory abnormalities than genotype IA.

S-510 A case of peritoneal dialysis-associated peritonitis caused by Agromyces mediolanus

( Ju Hwan Oh ),( Min Woo An ),( Hyun Kwon Ju ),( Na Yun Kang ),( A Young Cho ),( In O Sun ),( Kwang Young Lee ) 대한내과학회 2016 대한내과학회 추계학술대회 Vol.2016 No.1

Introduction:?Agromyces mediolanus (A.mediolanus) is an unusual pathogen in human infection. No case of peritoneal dialysis (PD)-associated peritonitis by A.mediolanus has been reported. This is the first case report of PD-associated peritonitis by A.mediolanus in the world. We describe a case of PD-associated peritonitis by A.mediolanus that was cured after catheter removal.?Case: A 59-year-old man who had been treated with CAPD was admitted to our hospital because of turbid peritoneal effluent accompanied by constant diffuse abdominal pain. The patient had an exit-site infection with purulent secretion. The laboratory findings showed PD peritonitis: the white blood cell (WBC) count of the peritoneal effluents was 1157/mm3 with a neutrophil predominance (91.3%). Cefazolin and ceftazidime were given intraperitoneally per day. The peritoneal WBC decreased, but the patient’s clinical condition didn’t improve on the 5th day after starting intraperitoneal antibiotics. Culture of the peritoneal dialysate revealed Agromyces species, which was susceptible to meropenem, imipenem, vancomycin, but resistant to ceftriaxone. Thus, we changed antibiotics to intraperitoneal vancomycin and meropenem on the 5th day. Despite the intraperitoneal antibiotics for 4 days, the patient’s abdominal pain persisted and the peritoneal WBC count became elevated. Therefore, we removed the PD catheter and the patient was switched to hemodialysis on the 9th day. Vancomycin and imipenem was continued for another 2 weeks and the patient’s clinical condition improved. The patient has been maintained on hemodialysis.?Conclusions: There is no previous report of peritonitis associated with A.mediolanus in a CAPD patient. In this case, we used intraperitoneal cefazolin and ceftazidime and changed to vancomycin and imipenem according to the result of peritoneal dialysate. However the patient improved after removal of catheter. Therefore, early removal of the PD catheter in A. mediolanus related peritonitis without prompt reaction to antibiotic therapy should be considered.

적외선열화상장치를 이용한 Buchanan plugger 표면의 온도상승 분석

최성아,김선호,황윤찬,윤창,오병주,최보영,정우남,정선와,황인남,오원만 大韓齒科保存學會 2002 Restorative Dentistry & Endodontics Vol.27 No.4

This study was performed to evaluate the temperature rise on various position of the Buchanan plugger, the peak temperature of plugger's type and the temperature change by its touching time of heat control spring. The heat carrier system 'System B' (Model 1005. Analytic Technologies, USA) and the Buchanan's pluggers of F, FM, M and ML sizes are used for this study. The temperature was set to 200℃ which Dr. Buchanan's "continuous wave of condensation" technique recommended on digital display and the power level on it was set to 10. In order to apply heat on the Buchanan's pluggers, the heat control spring was touched for 1, 2, 3, 4 and 5 seconds respectively. The temperature rise on the surface of the pluggers were measured at 0.5 mm intervals from tip to 20 mm length of shank using the infraed thermography (Radiation Thermometer-IR temper, NEC San-ei Instruments, Ltd, Japan) and TH31-702 Data capture software program (NEC San-ei Instruments, Ltd, Japan). Data were analyzed using a one way ANOVA followed by Duncan's multiple range test and linear regression test. The results as follows. 1. The position at which temperature peaked was approximately at 0.5 mm to 1.5 mm far from the tip of Buchanan's pluggers (p<0.001). The temperature was constantly decreased toward the shank from the tip of it(p<0.001). 2. When the pluggers were heated over 5 seconds, the peak temperature by time of measurement revealed from 253.3±10.5℃ to 192.1±3.3℃ in a touch for 1 sec, from 218.6±5.0℃ to 179.5±4.2℃ in a touch for 2 sec, from 197.5±3.0℃ to 167.6±3.7℃ in a touch for 3 sec, from 183.7±2.5℃ to 159.8±3.6℃ in a touch for 4 sec and from 164.9±2.0℃ to 158.4±1.8℃ in a touch for 5 sec. A touch for 1 sec showed the highest peak temperature, followed by, in descending order, 2 sec, 3 sec, 4 sec. A touch for 5 sec showed the lowest peak temperature (p<0.001). 3. A each type of pluggers showed different peak temperatures. The peak temperature was the highest in F type and followed by, in descending order, M type, ML type. FM type revealed the lowest peak temerature (p<0.001). The results of this study indicated that pluggers are designed to concentrate heat at around its tip, its actual temperature dose not correlate well with the temperature which Buchanan's "continuous wave of condensation" technique recommended, and finally a quick touch of heat control spring for 1 sec reveals the highest temperature rise.

으름 어린잎 식초의 품질특성과 생리활성

권우영(Woo-Young Kwon),이은경(Eun-Kyoung Lee),윤진아(Jin-A Yoon),정강현(Kang-Hyun Chung),이권재(Kwon-Jai Lee),송병춘(Byeong Chun Song),안정희(Jeung Hee An) 한국식품영양과학회 2014 한국식품영양과학회지 Vol.43 No.7

We investigated the characteristics and biological activities of vinegars added with different levels (0%, 0.5%, 1%, 2%, and 3%) of young leaves of Akebia quinata. During alcohol fermentation, alcohol and total acidity contents of vinegars increased. During acid fermentation, total acidity and amino acid contents increased. Vinegar added with 3% A. quinata leaf showed the highest total sensory score. The contents of total polyphenols, flavonoids, and tannin significantly increased during fermentation according to the amount of A. quinata leaf. After 22 days of fermentation, total polyphenol, total flavonoid, and tannin contents of vinegar added with 3% A. quinata were 4,079.08 mg GAE/100 g, 2,927.08 mg CE/100 g, and 3,618.00 mg TAE/100 g, respectively. ABTS radical scavenging activity of vinegar added with 3% A. quinata was 79.63%. Anti-cancer activities of vinegar added with 3% A. quinata were 48.65% and 52.90% against MCF-7 and HepG2 cells, respectively. Vinegar added with 3% A. quinata showed anti-bacterial activities against Bacillus cereus, Shigella flexneri, Salmonella enterica, Bacillus subtilis, and Klebsiella pneumoniae. Our results demonstrate that the biological activities of vinegar added with 3% A. quinata leaf (22 days of fermentation) were excellent, and their enhanced total polyphenol, flavonoid, and tannin contents were associated with antioxidant, anti-cancer and anti-microbial activities. Thus, A. quinata can be used as a functional material in vinegar and other foods.

Effect of Apoptosis Induction of Ailanthus altissima on Human Lung Carcinoma Cells

Yu-Jin Hwang,Hye-Im Woo,Inhye Kim,Dong-Sik Park,Jaehyun Kim,Ae-Son Om,Kyung-A Hwang 경상대학교 농업생명과학연구원 2011 농업생명과학연구 Vol.45 No.5

We investigated the inhibitory effects of solvent extracts from Ailanthus altissima in A549 human lung cancer cell. A. altissima has been recognized as a traditional healthy food due to its various biological activities against hypertension, strokes, fever, pain, neuralgia, inflammation, and cancer effects. Recently, it has been reported that the extracts of various wild vegetables show strong anti-cancer properties by induction of apoptosis. However, the mechanisms of their cytotoxicity in human lung cancer cells have been poorly understood. The present study was investigated the effects of solvent extracts from A. altissima on cell growth and apoptosis on A549 human lung cancer cells. A treatment of A. altissima inhibited the growth of A549 cells in a dose-dependent manner by inducing apoptosis. Especially, the chloroform fraction showed the highest anti-cancer effect among five kinds of fractions. And also, induction of apoptosis by chloroform fraction were associated with down-regulation of Bcl-2, and up-regulation of pro-apoptotic Bax expression. From these results, A. altissima may have a therapeutic potential in human lung cancer cells and as a functional food.

Sulforaphane exerts its anti-inflammatory effect against amyloid-β peptide via STAT-1 dephosphorylation and activation of Nrf2/HO-1 cascade in human THP-1 macrophages

An, Y.W.,Jhang, K.A.,Woo, S.Y.,Kang, J.L.,Chong, Y.H. Elsevier Science Pub. Co 2016 NEUROBIOLOGY OF AGING Vol.38 No.-

Alzheimer's disease (AD) is the most common neurodegenerative disorder worldwide, accounting for most cases of dementia in elderly individuals, and effective therapies are still lacking. This study was designed to investigate the anti-inflammatory properties of sulforaphane against Aβ<SUB>1-42</SUB> monomers in human THP-1 microglia-like cells. The results showed that sulforaphane preferentially inhibited cathepsin B- and caspase-1-dependent NLRP3 inflammasome activation induced by mostly Aβ<SUB>1-42</SUB> monomers, an effect that potently reduced excessive secretion of the proinflammatory cytokine interleukin-1β (IL-1β). Subsequent mechanistic studies revealed that sulforaphane mitigated the activation of signal transducer and activator of transcription-1 induced by Aβ<SUB>1-42</SUB> monomers. Sulforaphane also increased nuclear factor erythroid 2-related factor 2 (Nrf2) nuclear translocation, which was followed by upregulation of heme-oxygenase 1 (HO-1). The anti-inflammatory effect of sulforaphane on Aβ<SUB>1-42</SUB>-induced IL-1β production was diminished by small interfering RNA-mediated knockdown of Nrf2 or HO-1. Moreover, sulforaphane significantly attenuated the levels of microRNA-146a, which is selectively upregulated in the temporal cortex and hippocampus of AD brains. The aforementioned effects of sulforaphane were replicated by the tyrosine kinase inhibitor, herbimycin A, and Nrf2 activator. These results indicate that signal transducer and activator of transcription-1 dephosphorylation, HO-1 and its upstream effector, Nrf2, play a pivotal role in triggering an anti-inflammatory signaling cascade of sulforaphane that results in decreases of IL-1β release and microRNA-146a production in Aβ<SUB>1-42</SUB>-stimulated human microglia-like cells. These findings suggest that the phytochemical sulforaphane has a potential application in AD therapeutics.