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      • KCI등재

        해양세균 Agarivorans sp. BK-1의 분리 및 β-아가라제의 특성 규명

        안병기(Byeong-Ki Ahn),민경철(Kyung-Cheol Min),이동근(Dong-Geun Lee),김안드레(Andre Kim),이상현(Sang-Hyeon Lee) 한국생명과학회 2019 생명과학회지 Vol.29 No.11

        본 연구에서는 agarose를 분해하는 세균을 분리하고 한천분해효소의 특성을 조사하였다. 한천분해세균인 BK-1은 부산광역시 수영구 광안리해수욕장에서 채취한 바닷물 시료에서 Marine agar 2216 평판배지를 이용하여 분리하였다. 분리된 균은 16S rRNA 유전자 염기서열분석을 통해 Agarivorans sp. BK-1으로 명명하였다. 세포 외 agarase는 Agarivorans sp. BK-1 균주 배양액에서 획득하였으며, 이를 이용하여 효소의 특성을 조사하였다. Agarivorans sp. BK-1 균주의 한천분해효소는 20, 30, 40, 50, 60 및 70℃에서 각각 67, 93, 97, 100, 58, 52%의 상대활성을 나타냈으며, pH 5, 6, 7, 8에서는 59, 100, 95, 91%의 활성을 나타내었다. 세포 외 agarase는 50℃에서 pH 6.0인 20 mM Tris-HCl 완충용액을 사용하였을 때 최대의 활성을 보였다. 이 agarase는 20, 30, 40℃에서 2시간 동안 열처리하여도 90% 이상의 잔존활성을 보였다. 또한, 50℃에서 2시간 동안 노출된 후에도 56%의 잔존활성을 보였다. 60과 70℃에서 30분 동안 노출된 후에는 효소활성 대부분이 사라졌다. Zymogram 분석을 통하여 Agarivorans sp. BK-1 균주가 약 110, 90, 55 kDa 크기의 한천분해효소들을 생산하는 것을 확인할 수 있었다. TLC 분석 결과, Agarivorans sp. BK-1 균주의 한천분해효소는 네오한천올리고당인 neoagarobiose (46.8%), neoagarotetraose (39.7%) 및 neoagarohexaose (13.5%)를 생성하는 것으로 보아 β-agarase로 확인되었다. 따라서 Agarivorans sp. BK-1가 생산하는 β-agarase는 보습효과, 세균성장 억제, 미백효과, 식품, 화장품 및 전분노화 방지 등의 기능을 가지는 네오한천올리고당의 생산에 유용할 것으로 판단된다. The purpose of this study was to isolate an agar-degrading marine bacterium and characterize its agarase. Bacterium BK-1, from Gwanganri Beach at Busan, Korea, was isolated on Marine 2216 agar medium and identified as Agarivorans sp. BK-1 by 16S rRNA gene sequencing. The extracellular agarase, characterized after dialysis of culture broth, showed maximum activity at pH 6.0 and 50℃ in 20 mM Tris-HCl buffer. Relative activities at 20, 30, 40, 50, 60, and 70℃ were 67, 93, 97, 100, 58, and 52%, respectively. Relative activities at pH 5, 6, 7, and 8 were 59, 100, 95, and 91%, respectively. More than 90% of the activity remained after a 2 hr exposure to 20, 30, or 40℃; about 60% of the activity remained after a 2 hr exposure to 50℃. Almost all activity was lost after exposure to 60 or 70℃ for 30 min. Zymography revealed three agarases with molecular weights of 110, 90, and 55 kDa. Agarose was degraded to neoagarobiose (46.8%), neoagarotetraose (39.7%), and neoagarohexaose (13.5%), confirming the agarase of Agarivorans sp. BK-1 as a β-agarase. The neoagarooligosaccharides generated by this agarase could be used for moisturizing, bacterial growth inhibition, skin whitening, food treatments, cosmetics, and delaying starch degradation.

      • KCI등재

        족삼리자침(足三里刺鍼)이 가토소장(家兎小腸)의 운동성(運動性)에 미치는 영향(影響)

        이방성,윤현민,장경전,송춘호,안창범,Lee, Bang-Sung,Youn, Hyoun-Min,Jang, Kyung-Jeon,Song, Choon-Ho,Ahn, Chang-Beohm 대한침구의학회 2000 대한침구의학회지 Vol.17 No.2

        This study was undertaken to determine effect of Zusan1i(ST36) acupuncture (AP) on the small intestinal motility in the rabbit. Changes in the motility in vivo of ileum was measured before and 20 min. after AP. In order to examine whether AP effect was affected by autonomic nerves, norepinephrine or acetylcholine was applicated. AP significantly increased the mean wave amplitude and motor index, but not the mean wave duration and motor activity. Infusion of norepinephrine ($2{\mu}m$) into an ear vein resulted in a decrease in the mean wave amplitude and motor index, which was not affected by AP. Acetylcholine ($10{\mu}m$) produced inhibition followed by stimulation of motility. The stimulation of motility was induced by an increase in the mean wave amplitude and motor index, but not the mean wave duration and motor activity. The effect of acetylcholine was not significantly different between before and after AP. These results suggest that Zusan1i(ST36) acupuncture stimulates the small intestinal motility, which is not mediated by autonomic nerves. Therefore, Zusan1i(ST36) acupuncture seems to be applicable to impaired small intestinal movement.

      • KCI등재

        요통(腰痛) 환자에 대한 침치료(針治療)와 직접구(直接灸) 병행치료(竝行治療)에 대한 비교(比較) 연구(硏究)

        위종성,원승환,황정수,선승호,안영민,손승현,박기철,박희수,We, Jong-sung,Won, Seung-hwan,Hwang, Jung-soo,Sun, Seung-ho,Ahn, Young-min,Sohn, Seung-hyun,Park, Ki-chul,Park, Hee-soo 대한침구의학회 2004 대한침구의학회지 Vol.21 No.6

        Objective : The purpose of this report is to examine the effects of direct moxibustion in the Low back pain patient. Methods : Clinical studies were done 30 patients who were treated with low back pain to Dept. of Acupuncture & Moxibustion, of Oriental Medical Sang-Ji University from September 1, 2003 to August 31, 2004. We treated them by Oriental medical therapy(including direct moxibustion)for 10 days. The evaluation was performed five times(admission day, before and after each twice). Results : 1. VAS was decreased after direct moxibustion for 10 days significantly(p<0.01). 2. After direct moxibustion, S.L.R. T angle of patients were increased. ConclUsions : We brought to the conclusion that direct moxibustion has possibility to the efficient to cure the Low back pain. So we suggest the possibility to use this treatment for Low back pain.

      • KCI등재후보

        Interaction between Neuronal Depolarization and MK-801 in SH-SY5Y Cells and the Rat Cortex

        김예니,Miran Seo,이윤일,So-Young Kim,Eun-Ah Cho,김세현,Yong-Min Ahn,Ung-Gu Kang,김용식,Yong-Sung Juhnn 대한신경정신의학회 2008 PSYCHIATRY INVESTIGATION Vol.5 No.2

        Objective: The interaction between MK-801, a model of psychosis and KCl-induced depolarization or electroconvulsive shock (ECS), a therapeutic model of electroconvulsive therapy (ECT), was investigated in SH-SY5Y cells and the rat frontal cortex. Methods: SH-SY5Y cells were pretreated with 1 μM MK-801 for 15 min, followed by cotreatment with 100 mM KCl for 5 min. MK-801 was reintroduced after the KCl was washed out, and the samples were incubated before harvesting. For the experiments in rats, male Sprague-Dawley rats were treated with MK-801 followed by ECS. Immunoblot analyses of glycogen synthase kinase 3β (GSK3β) (Ser9), AKT (Ser473) and extracellular legulated kinase (ERK)1/2 in SH-SY5Y cells and the rat frontal cortex were performed. Results: KCl-induced neuronal depolarization resulted in the transient dephosphorylation of AKT (Ser473) and GSK3β (Ser9), followed by increased phosphorylation of the enzymes in SH-SY5Y cells. Cotreatment with MK-801 and KCl inhibited the initial dephosphorylation of AKT and GSK3β produced by KCl-induced neuronal depolarization. Similarly, ECS resulted in the transient dephosphorylation of AKT (Ser473) and GSK3β (Ser9), whereas cotreatment with MK-801 inhibited the initial dephosphorylation of AKT (Ser473) and GSK3β (Ser9) produced by ECS in the rat frontal cortex. No significant interaction was observed between MK-801 and KCl in the dephosphorylation of ERK1/2. Conclusion: These results suggest that an antagonistic interplay between MK-801 and neuronal depolarization by KCl or ECS is involved the regulation of AKT (Ser473) and GSK3β (Ser9) phosphorylation.

      • 피프로닐 오염도 조사를 위한 분석법 정립

        조순길 ( Soon-kil Cho ),이미진 ( Mi-jin Lee ),이정민 ( Jeong-min Lee ),김호진 ( Ho-jin Kim ),박수민 ( Soo-min Park ),박혜진 ( Hye-jin Park ),이승화 ( Seung-hwa Lee ),안종성 ( Jong-sung Ahn ) 한국환경농학회 2018 한국환경농학회 학술대회집 Vol.2018 No.-

        2017년 유럽에서 피프로닐에 오염된 계란이 유통된데 이어 국내에서 생산된 일부 계란에서도 살충제가 검출되었다. 피프로닐은 진드기 방제 목적으로 사용되는 살충제로 sulfide, sulfone, amide, sulfoxide의 대사산물을 생성한다. 피프로닐은 물에 1.9 mg/L 정도 용해되고, 아세톤에 545.9g/L가 용해되는 성질이 있어 시료 채취시 아세톤을 사용한다. 농장시설 등에 부착된 피프로닐을 제거하기 위해 5% 탄산나트륨(Na2CO3)과 15% 과산화수소(H2O2)를 권장하고 있다. 이는 피프로닐을 피프로닐설폰으로 산화시킨 다음 반복적으로 사용하여 제거한다. 이러한 제거 작업을 통해 피프로닐이 제거된 정도를 조사할 필요가 있고, 이를 위한 분석법 정립이 필요하다. 시료를 250 mL 용기에 넣고 1% 함유 아세토니트릴 100 mL을 주가하여 10분간 격렬하게 진탕한 후, 3500 rpm으로 원심분리를 하였다. 추출액 1 mL을 0.2 um 멤브레인필터로 여과하여 시험용액으로 하였다. 칼럼은 C18(2.0 mm I.d×50mm, 3 um)을 사용하였고, 이동상은 0.1% 포름산 함유 메탄올과 0.1% 포름산 및 5 mM 아세트산암모늄이 함유된 물을 사용하였다. 분석조건으로 피프로닐의 선구이온은 437 m/z, 생성이온은 368, 255, 290 m/z, positive 조건으로 분석하였고, 피프로닐 설폰의 선구이온은 451 m/z, 생성이온은 415, 282, 244 m/z negative 조건으로 분석하였다. 분석법의 정량한계는 피프로닐 0.92 ug/kg, 피프로닐설폰 0.70 ug/kg이었다. 피프로닐과 피프로닐 설폰을 각각 1, 5, 10, 50, 100 ug/kg 수준으로 검량선을 작성한 결과, r2는 모두 0.9999이었다. 10 ug/kg과 50 ug/kg 수준의 3반복 회수율 실험결과, 10ug/kg 수준에서 피프로닐은 74.7±0.56%, 피프로닐 설폰은 88.9± 0.27%로 나타났다. 50 ug/kg 수준에서는 피프로닐은 78.6±0.11%, 피프로닐 설폰은 95.2±0.02%로 유효회수율 범위(70~120%)에 포함되었다. 머무름 시간은 피프로닐 5.3 mim, 피프로닐 설폰은 5.7min에서 검출되었다. 교차확인 목적으로 LC/MS/MS로 분석한 시료를 GC/μ-ECD로 분석을 하였다. 분석조건은 주입구 온도 250℃, (5%-Phenyl)-methylpolysiloxane이 충진된 30 m×0.25 mm×0.25 um길이의 칼럼이다. 오븐온도는 130℃에서 300℃까지 승온 조건으로 분석하였고, 머무름시간은 피프로닐 11.64 min, 피프로닐 설폰12.79 min으로 LC/MS/MS와 GC/μ-ECD 모두 표준물질과 시료의 결과가 일치함을 알 수 있었다. 정립된 분석법은 피프로닐 오염도조사를 위한 분석법으로 활용할 수 있을 것으로 판단된다.

      • KCI등재후보

        척수손상 흰 쥐의 운동기능 회복에 미치는 손상부위 직접자극을 통한 기능적 자기자극치료 효과

        조윤우,김수정,박해운,서정민,황세진,장성호,이동규,안상호,Cho, Yun-Woo,Kim, Su-Jeong,Park, Hea-Woon,Seo, Jeong-Min,Hwang, Se-Jin,Jang, Sung-Ho,Lee, Dong-Gyu,Ahn, Sang-Ho 대한물리치료학회 2011 대한물리치료학회지 Vol.23 No.1

        Purpose: The purpose of this study was to determine the effect of direct functional magnetic stimulation (FMS) of affected spinal cord on motor recovery following spinal cord injury in rats. Methods: After a contusion injury at the spinal level T9 using an NYU Impactor, functional magnetic stimulation was delivered by a magnetic stimulator through a round prototype coil (7 cm in diameter). Stimulation parameters were set as follows: repetition rate = 50 Hz (stimulus intensity 100% = 0.18 T), stimulation time = 20 min. Functional magnetic stimulation was administered twice a day, 5 days per week for 8 weeks starting 4 days after spinal cord injury. Functional magnetic stimulationwas delivered directly to the affected spinal cord. Outcomes of locomotor performance were assessed by the Basso Beattie Bresnahan (BBB) locomotor rating scale and by an inclined plane test weekly for 8 weeks. Results: In the BBB test, hindlimb motor function in the Functional magnetic stimulation group improved significantly more compared to the control group at 3, 4, 6, 7, and 8 weeks (p<0.05). In the inclined plane test, the angle of the plane in the functional magnetic stimulation group increased significantly more compared to the control group at 4, 5, 7, and 8 weeks (p<0.05). Conclusion: Our results demonstrate that direct Functional magnetic stimulation of the lesional site may have beneficial effects on motor improvement after spinal cord injury.

      • KCI등재
      • KCI등재

        Chronic allograft injury by subclinical borderline change

        Sang-IL Min,Young Suk Park,Sanghyun Ahn,Taejin Park,Dae Do Park,Suh Min Kim,Kyung Chul Moon,Seung-Kee Min,Yon Su Kim,Curie Ahn,Sang Joon Kim,Jongwon Ha 대한외과학회 2012 Annals of Surgical Treatment and Research(ASRT) Vol.83 No.6

        Purpose: This study investigated the impact of subclinical borderline changes on the development of chronic allograft injury in patients using a modern immunosuppression protocol. Methods: Seventy patients with stable renal allograft function and who underwent protocol biopsies at implantation, 10 days and 1 year after transplantation were included and classified based on biopsy findings at day 10. The no rejection (NR) group included 33 patients with no acute rejection. The treatment (Tx) group included 21 patients with borderline changes following steroid pulse therapy, and the nontreatment (NTx) group included 16 patients with borderline changes nontreated. Results: The Banff Chronicity Score (BChS) and modified BChS (MBChS) were not different among the three groups at implantation (P = 0.48) or on day 10 (P = 0.96). Surprisingly, the NTx group had more prominent chronic scores at the 1-year biopsy, including BChS (3.07 ± 1.33, P = 0.005) and MBChS (3.14 ± 1.41, P = 0.008) than those in the Tx and NR group, and deterioration of BChS was more noticeable in the NTx group (P = 0.037), although renal function was stable (P = 0.66). No difference in chronic injury scores was observed between the Tx and NR groups at the 1-year biopsy. Conclusion: Subclinical borderline changes can be a risk factor for chronic allograft injury and should be considered for antirejection therapy.

      • SCISCIESCOPUS

        Tat-ATOX1 inhibits streptozotocin-induced cell death in pancreatic RINm5F cells and attenuates diabetes in a mouse model

        AHN, EUN HEE,KIM, DAE WON,SHIN, MIN JEA,RYU, EUN JI,YONG, JI IN,CHUNG, SEOK YOUNG,CHA, HYUN JU,KIM, SANG JIN,CHOI, YEON JOO,KIM, DUK-SOO,CHO, SUNG-WOO,LEE, KEUNWOOK,CHO, YOON SHIN,KWON, HYEOK YIL,PARK UNKNOWN 2016 INTERNATIONAL JOURNAL OF MOLECULAR MEDICINE Vol.38 No.1

        <P>Antioxidant 1 (ATOX1) functions as an antioxidant against hydrogen peroxide and superoxide, and therefore may play a significant role in many human diseases, including diabetes mellitus (DM). In the present study, we examined the protective effects of Tat-ATOX1 protein on streptozotocin (STZ)-exposed pancreatic insulinoma cells (RINm5F) and in a mouse model of STZ-induced diabetes using western blot analysis, immunofluorescence staining and MTT assay, as well as histological and biochemical analysis. Purified Tat-ATOX1 protein was efficiently transduced into RINm5F cells in a dose-and time-dependent manner. Additionally, Tat-ATOX1 protein markedly inhibited reactive oxygen species (ROS) production, DNA damage and the activation of Akt and mitogen activated protein kinases (MAPKs) in STZ-exposed RINm5F cells. In addition, Tat-ATOX1 protein transduced into mice pancreatic tissues and significantly decreased blood glucose and hemoglobin A1c (HbA1c) levels as well as the body weight changes in a model of STZ-induced diabetes. These results indicate that transduced Tat-ATOX1 protein protects pancreatic beta-cells by inhibiting STZ-induced cellular toxicity in vitro and in vivo. Based on these findings, we suggest that Tat-ATOX1 protein has potential applications as a therapeutic agent for oxidative stress-induced diseases including DM.</P>

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