• Korean Chemical Engineering Research
• /
• 제43권2호
• /
• pp.187-201
• /
• 2005

최근 전통적인 액체상 공정을 대체하는 기술로서 고체 담체와 단백질 사이의 '생물인식' 기능을 이용하는 새로운 생물공정기술이 개발되고 있다. 통상 고체 담체로는 표면에 특정한 기능기가 노출되어 있는 크로마토그래피용 담체를 사용한다. 단백질의 반응이나 상호작용이 단백질이 담체 표면에 부착되어 있는 상태에서 일어나기 때문에 이 '고체상 기술'은 액체상 기술에 비해 뚜렷한 장점을 갖고 있다. 고체상 재접힘은 변성제에 의해 용해된 내포체 형태의 재조합 단백질을 이온교환수지 표면에 흡착시켜 시작한다. 변성제를 단백질 주위로부터 서서히 제거시키면서 고유의 3차 구조로 재접힘시킨다. 재접힘이 완료되면 염 구배와 같은 전통적인 방법에 의해 재접힘된 단백질을 정제된 상태로 용출시킨다. 이 개념은 '확장층 흡착 재접힘'에도 연장 적용된다. 세포파쇄액에 변성제를 첨가하여 용해한 내포체 단백질은 확장층 흡착 크로마토그래피용 Streamline 담체에 흡착되고 세포찌꺼기와 불순 단백질들은 확장층 사이로 빠져 칼럼 밖으로 제거된다. 흡착된 목적 단백질은 고체상 재접힘 방법에 의해 재접힘 된 후 용출된다. 수년간 연구 발전되어 온이 새로운 재접힘 기술은 정제수율 향상, 공정 단계 감축, 공정 시간 및 부피 감소에 따라 생물의약공정의 경제성을 크게 향상시킬 수 있는 것으로 증명되고 있다. 본 논문에서는 실험실에서 수행한 여러 생물의약용 단백질들을 대상으로 한 연구 실험 자료를 바탕으로 고체상 재접힘 기술의 적용 사례를 서술하였다.

• Molecules and Cells
• /
• 제37권11호
• /
• pp.833-840
• /
• 2014

Cullin4-RING ubiquitin ligase (CRL4) is a family of multi-subunit E3 ligases. To investigate the possible involvement of CRL4 in heat stress response, we screened T-DNA insertion mutants of putative CRL4 substrate receptors that exhibited altered patterns in response to heat stress. One of the mutants exhibited heat stress tolerance and was named heat stress tolerant DWD1 (htd1). Introduction of HTD1 gene into htd1-1 led to recovery of heat sensitivity to the wild type level, confirming that the decrease of HTD1 transcripts resulted in heat tolerance. Therefore, HTD1 plays a negative role in thermotolerance in Arabidopsis. Additionally, HTD1 directly interacted with DDB1a in yeast two-hybrid assays and associated with DDB1b in vivo, supporting that it could be a part of a CRL4 complex. Various heat-inducible genes such as HSP14.7, HSP21, At2g03020 and WRKY28 were hyper-induced in htd1-1, indicating that HTD1 could function as a negative regulator for the expression of such genes and that these genes might contribute to thermotolerance of htd1-1, at least in part. HTD1 was associated with HSP90-1, a crucial regulator of thermotolerance, in vivo, even though the decrease of HTD1 did not affect the accumulation pattern of HSP90-1 in Arabidopsis. These findings indicate that a negative role of HTD1 in thermotolerance might be achieved through its association with HSP90-1, possibly by disturbing the action of HSP90-1, not by the degradation of HSP90-1. This study will serve as an important step toward understanding of the functional connection between CRL4-mediated processes and plant heat stress signaling.

• 한방재활의학과학회지
• /
• 제18권2호
• /
• pp.61-79
• /
• 2008

Objectives : The purpose of this study was to identify the effect of Dokwal-tang(Duhuo-tang) and Jungsongouhyul Pharmacopuncture on pain control and nerve regeneration after crush injury in rat sciatic nerve. Methods : Animal model was produced through right sciatic nerve crush injury and they were randomly divided into four groups; Group I: no treatment control group; Group II: experimental group treated with Dokwal-tang(Duhuo-tang); Group III: experimental group treated with Jungsongouhyul Pharmacopuncture; Group IV: experimental group treated with Dokwal-tang(Duhuo-tang) and Jungsongouhyul Pharmacopuncture. For the pain assessment, this study was observed the paw withdrawal latency(PWL) and immunoreactivity on the substance-P. For the nerve regeneration assessment, the sciatic functional index(SFI) and immunoreactivity on the GAP-43 were measured. Results : 1. In the pain assessment, the PWL of experimental groups were significantly higher than control group and group IV was significantly higher than other groups. 2. In immunohistochemical response of substance-P, as time passes, all groups had decreased immunoreactivity gradually. Specially, group IV was observed the lowest immunoreactivity. 3. In the assessment of SFI, the SFI of experimental groups were significantly higher than control group. 4. In immunohistochemical response of GAP-43, all groups had higher GAP-43 immunoreactivity at the 14 days from post-injury and at the all days, control group was observed most lower immunoreactivity and group IV was observed most higher immunoreactivity. Conclusions : These results of this study suggest that Dokwal-tang(Duhuo-tang) and Jungsongouhyul Pharmacopuncture was related with pain inhibition and motor nerve recovery, and decreased substance-P expression and increased GAP-43 expression after sciatic nerve crush injury.

• 한국산학기술학회논문지
• /
• 제17권5호
• /
• pp.522-529
• /
• 2016

본 연구는 전방십자인대 재건술을 시행한 환자를 대상으로 후방보행을 포함한 재활운동 프로그램이 환자의 통증지수와 ROM, 고유수용성감각에 어떠한 영향을 미치는지 구명하고자 하였다. 이 연구의 대상자는 전방십자인대 손상으로 재건술을 시행한 환자 14명으로 하였으며 후방보행 집단(n=7)과 통제집단(n=7)으로 무선배정 하였고 4주간 각각의 재활운동프로그램을 실시하였다. 이 연구의 결과에서, 굴곡 관절가동범위는 운동 전 후, 후방보행 집단(p<.001)과 통제집단(p<.01) 모두 운동 후 유의하게 감소되었으며 신전 관절가동범위에서는 후방보행 집단(p<.05)에서만 운동 후 유의한 차이를 나타내었고 두 변인 모두 집단 간의 차이는 보이지 않았다. 통증지수에 대한 결과에서는 운동 후, 두 집단 모두 유의하게 감소되었고(p<.001) 집단 간 차이에서는 운동 후, 후방보행 집단이 통제집단보다 유의하게 낮은 것으로 나타났으며(p<.001), 고유수용성감각에서는 후방보행 집단(p<.001)과 통제집단(p<.05) 모두 운동 후 유의하게 감소하였으나 집단 간의 차이는 나타나지 않았다. 이러한 결과는 후방보행의 재활운동이 전방십자인대 재건술을 시행한 환자의 통증을 감소시키고 관절의 가동범위 확보와 고유수용성감각을 회복하는데 긍정적인 효과가 있는 것으로 사료된다.

• Journal of Korean Neurosurgical Society
• /
• 제30권2호
• /
• pp.194-200
• /
• 2001

Object : Anterior cervical discectomy and fusion is accepted as appropriate surgical intervention for disease processes of the ventral cervical spine. In some cases, however, improvement of symptom following operation develop myelopathy or rhizopathy at longterm follow-up. We studied to clarify the correlation between clinical results and plain radiologic findings at long term follow-up. Methods : A total of 86 patients who underwent anterior cervical discectomy and fusion were grouped into deteriorated and good recovery group. These two groups were compared in lateral functional roentgenograms on the cervical kyphosis and disc height at fused level, sagittal plane rotation and dynamic sagittal canal diameter at adjacent level. Results : The presence of cervical kyphosis and disc height at fused level do not correlate with long term followup results. Sagittal plane rotation of more than 20 degrees were identified in 36% of the cases in deteriorated group, whereas the same findings were identified in 15% of the cases in good group. Sagittal diameter of less than 12mm were identified in 48% of the cases in deteriorated group, whereas the same finding were identified in 8% of the cases in good group. Preoperatively, sagittal plane rotation of more than 20 degrees at adjacent level were identified in 28% of the cases in deteriorated group, whereas the same findings were identified in 13% of the cases in good group. Preoperatively sagittal diameter of less than 12mm at adjacent level were identified in 40% of the cases in deteriorated group, whereas the same finding were identified in 7% of the cases in good group. Conclusion : Large sagittal plane rotation and small dynamic sagittal diameter at adjacent level were factors that might be associated with later clinical deterioration after cervical anterior fusion.

• 대한응급의학회지
• /
• 제28권5호
• /
• pp.457-466
• /
• 2017

Purpose: Traumatic brain injury (TBI) is a significant cause of morbidity and mortality worldwide. Severity of the initial insult is one of the most significant factors affecting outcome following TBI. In order to investigate the mechanisms of cellular injury and develop novel therapeutic strategies for TBI, we designed a standardized animal TBI model and evaluated histological and functional outcomes according to the degree of impact severity. Methods: Male adult C57Bl/6 mice underwent controlled cortical impact (CCI) at varying depths of deflection (1.0-2.0 mm). We performed hematoxylin and eosin staining at 7 days after recovery from TBI. Neurobehavioral characterization after TBI was analyzed by the Barnes maze test, passive avoidance test, open field test, rotarod test, tail suspension test, and light/dark test. Results: We observed a graded injury response according to the degree of deflection depths tested (diameter, 3 mm; velocity, 3 m/s; and duration, 500 ms) compared to sham controls. In the Barnes maze test, the severe TBI (2 mm depth) group showed reduced spatial memory as compared with the sham and mild TBI (1 mm depth) groups at 7 days after TBI. There was a significant difference in the results of the open field test and light/dark test among the three groups. Conclusion: Our findings demonstrate that the graded injury responses following TBI resulted in differential histopathological and behavioral outcomes in a mouse experimental CCI model. Thus, a model of CCI with histologic/behavioral outcome analysis may offer a reliable and convenient design for preclinical TBI research involving mice.

• Asian-Australasian Journal of Animal Sciences
• /
• 제32권11호
• /
• pp.1776-1788
• /
• 2019

Objective: The demands for measures to improve disease resistance and productivity of livestock are increasing, as most countries prohibit the addition of antibiotics to feed. This study therefore aimed to uncover functional feed additives to help enhance livestock immunity and disease resistance, using Acanthopanax sessiliflorus fruit extract (ASF). Methods: ASF was extracted with 70% EtOH, and total polyphenolic and catechin contents were measured by the Folin-Ciocalteu and vanillin assay, respectively. The 3D4/31 porcine macrophage cells ($M{\Phi}$) were activated by phorbol 12-myristate 13-acetate (PMA), and cell survival and growth rate were measured with or without ASF treatment. Flow-cytometric analysis determined the lysosomal activity, reactive oxygen species levels (ROS), and cell cycle distribution. Nuclear factor kappa B ($NF-{\kappa}B$) and superoxide dismutase (SOD) protein expression levels were quantified by western blotting and densitometry analysis. Quantitative polymerase chain reaction was applied to measure the lipid metabolism-related genes expression level. Lastly, the antibacterial activity of 3D4/31 $M{\Phi}$ cells was evaluated by the colony forming unit assay. Results: ASF upregulated the cell viability and growth rate of 3D4/31 $M{\Phi}$, with or without PMA activation. Moreover, lysosomal activity and intracellular ROS levels were increased after ASF exposure. In addition, the antioxidant enzyme SOD2 expression levels were proportionately increased with ROS levels. Both ASF and PMA treatment resulted in upregulation of $NF-{\kappa}B$ protein, tumor necrosis factor $(TNF){\alpha}$ mRNA expression levels, lipid synthesis, and fatty acid oxidation metabolism. Interestingly, co-treatment of ASF with PMA resulted in recovery of $NF-{\kappa}B$, $TNF{\alpha}$, and lipid metabolism levels. Finally, ASF pretreatment enhanced the in vitro bactericidal activity of 3D4/31 $M{\Phi}$ against Escherichia coli. Conclusion: This study provides a novel insight into the regulation of $NF-{\kappa}B$ activity and lipid metabolism in $M{\Phi}$, and we anticipate that ASF has the potential to be effective as a feed additive to enhance livestock immunity.

• Journal of Ginseng Research
• /
• 제37권4호
• /
• pp.401-412
• /
• 2013

Korean Red Ginseng (KRG) is an oriental herbal preparation obtained from Panax ginseng Meyer (Araliaceae). To expand our understanding of the action of KRG on central nervous system (CNS) function, we examined the effects of KRG on tissue plasminogen activator (tPA)/plasminogen activator inhibitor-1 (PAI-1) expression in rat primary astrocytes. KRG extract was treated in cultured rat primary astrocytes and neuron in a concentration range of 0.1 to 1.0 mg/mL and the expression of functional tPA/PAI-1 was examined by casein zymography, Western blot and reverse transcription-polymerase chain reaction. KRG extracts increased PAI-1 expression in rat primary astrocytes in a concentration dependent manner (0.1 to 1.0 mg/mL) without affecting the expression of tPA itself. Treatment of 1.0 mg/mL KRG increased PAI-1 protein expression in rat primary astrocytes to $319.3{\pm}65.9%$ as compared with control. The increased PAI-1 expression mediated the overall decrease in tPA activity in rat primary astrocytes. Due to the lack of PAI-1 expression in neuron, KRG did not affect tPA activity in neuron. KRG treatment induced a concentration dependent activation of PI3K, p38, ERK1/2, and JNK in rat primary astrocytes and treatment of PI3K or MAPK inhibitors such as LY294002, U0126, SB203580, and SP600125 (10 ${\mu}M$ each), significantly inhibited 1.0 mg/mL KRG-induced expression of PAI-1 and down-regulation of tPA activity in rat primary astrocytes. Furthermore, compound K but not other ginsenosides such as Rb1 and Rg1 induced PAI-1 expression. KRG-induced up-regulation of PAI-1 in astrocytes may play important role in the regulation of overall tPA activity in brain, which might underlie some of the beneficial effects of KRG on CNS such as neuroprotection in ischemia and brain damaging condition as well as prevention or recovery from addiction.

• 한국식품위생안전성학회지
• /
• 제36권5호
• /
• pp.376-381
• /
• 2021

Red beet roots (Beta vulgaris L.)는 천연색소로 붉은 색계열의 betacyanins은 75-95%의 betanine와 이성질체인 isobetanine 15-45%으로 존재한다. 본 연구는 비트레드를 사용한 식품에 대해 HPLC-DAD를 이용하여 지표성분인 betanine 및 isobetanine에 대해 분석법을 확립하였으며, 유효성 검증을 위해 직선성, 검출한계, 정량한계, 정확성, 정밀성, 측정불확도를 측정하였다. 캔디류, 빙과류, 코코아 가공품의 matrix에 적용하여 matrix matched calibration법을 사용하였으며 R²이 0.9998 이상으로 높은 직선성을 보였다. 검출한계와 정량한계는 각각 0.16-0.32 mg/L, 0.48-0.97 mg/L으로 확인되었다. 분석법의 정확성 및 정밀성을 검증하기 위해 intra-day 및 inter-day 반복 실험 결과, 회수율은 96.0-103.1 %, 100.0-102.2 %이였으며, RSD는 0.5-3.3 %, 0.9-3.8 %로 산출되었다. 측정불확도는 매트릭스 및 측정 농도에 따라 1.71-12.43%로 평가되었다. 또한, 확립된 분석법의 적용성 검토를 위해서 비트레드 색소를 사용한 가공식품 26종을 분석한 결과, betanine과 isobetanine을 정량 할 수 있었다 (8.4-3,823.4 mg/kg).

• 한국항해항만학회지
• /
• 제42권6호
• /
• pp.478-485
• /
• 2018

항로표지는 해상교통의 안전을 도모하고 선박운항 능률 향상을 위하여 설치된 해양교통안전시설 중 하나이다. 그러나 선박충돌 또는 기상악화 등의 원인으로 발생하는 항로표지의 사고로 인하여 제 기능을 상실 하였을 경우, 선박운항자를 비롯한 여러 이용자에게 이용불편 또는 항행위험을 증가시키고 있다. 특히 우리나라 최대 컨테이너항만인 부산항은 최근 12년(2006년~2017)동안 총400건의 항로표지 사고가 발생하였으며, 이로 인하여 평균 20.6일의 복구기간이 소요됨에 따라 항로표지 이용자들에게 해양안전 불안을 가중시키고 있다. 따라서 연구의 목적은 우리나라 항로표지 사고예방과 효율적인 관리방안을 적극적으로 추진하기 위한 이용자들의 정량적 지지도를 제시하고자 한다. 이를 위해 부산항 해역이용자들을 대상으로 항로표지 및 항로표지 사고에 대한 인식조사를 실시하고, 효율적 항로표지 관리방안의 추진을 통한 항행위험 감소에 대한 경제적 가치를 추정하였다. 연구의 결과, 비시장재화인 환경재나 공공재의 대표적인 가치추정법인 조건부가치추정법을 이용하여 추정된 부산항 항로표지 서비스에 대하여 이용자들이 부여한 경제적 가치는 최소 약 160억원으로 나타났다. 따라서 이러한 연구결과는, 항로표지 관리자 및 정책입안자들에게 부산항을 비롯한 우리나라 항로표지 서비스에 대한 다양한 이용자의 요구사항을 확인하고, 이에 대응하는 보다 효율적인 관리방안의 수립과 추진에 유용한 자료로 활용될 수 있을 것이다.