• Macromolecular Research
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• 제11권5호
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• pp.334-340
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• 2003

In order to fabricate new sustained delivery device of nerve growth factor (NGF), we developed NGF-loaded biodegradable poly(L-lactide-co-glycolide) (PLGA, the mole ratio of lactide to glycolide 75:25, molecular weight: 83,000 and 43,000 g/mole, respectively) film by novel and simple sandwich solvent casting method for the possibility of the application of neural tissue engineering. PLGA was copolymerized by direct condensation reaction and the molecular weight was controlled by reaction time. Released behavior of NGF from NGF-loaded films was characterized by enzyme linked immunosorbent assay (ELISA) and degradation characteristics were observed by scanning electron microscopy (SEM) and gel permeation chromatography (GPC). The bioactivity of released NGF was identified using a rat pheochromocytoma (PC-12) cell based bioassay. The release of NGF from the NGF-loaded PLGA films was prolonged over 35 days with zero-order rate of 0.5-0.8 ng NGF/day without initial burst and could be controlled by the variations of molecular weight and NGF loading amount. After 7 days NGF released in phosphate buffered saline and PC-12 cell cultured on the NGF-loaded PLGA film for 3 days. The released NGF stimulated neurite sprouting in cultured PC-12 cells, that is to say, the remained NGF in the NGF/PLGA film at 37 $^{\circ}C$ for 7 days was still bioactive. This study suggested that NGF-loaded PLGA sandwich film is released the desired period in delivery system and useful neuronal growth culture as nerve contact guidance tube for the application of neural tissue engineering.

• 대한약침학회지
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• 제12권1호
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• pp.67-76
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• 2009

Objective : Following central nervous system(CNS) injury, inhibitory influences at the site of axonal damage occur. Glial cells become reactive and form a glial scar, gliosis. Also myelin debris such as MAG inhibits axonal regeneration. Astrocyte-rich gliosis relates with up-regulation of GFAP and CD81, and eventually becomes physical and mechanical barrier to axonal regeneration. MAG is one of several endogenous axon regeneration inhibitors that limit recovery from CNS injury and disease. It was reported that molecules that block such inhibitors enhanced axon regeneration and functional recovery. Recently it was reported that treatment with anti-CD81 antibodies enhanced functional recovery in the rat with spinal cord injury. So in this current study, the author investigated the effect of the water extract of Uncariae Ramulus et Uncus on the regulation of CD81, GFAP and MAG that increase when gliosis occurs. Methods : MTT assay was performed to examine cell viability, and cell-based ELISA, western blot and PCR were used to detect the expression of CD81, GFAP and MAG. Then also immunohistochemistry was performed to confirm in vivo. Results : Water extract of Uncariae Ramulus et Uncus showed relatively high cell viability at the concentration of 0.05%, 0.1% and 0.5%. The expression of CD81, GFAP and MAG in astrocytes was decreased after the administration of Uncariae Ramulus et Uncus water extract. These results was confirmed in the brain sections following cortical stab injury by immunohistochemistry. Conclusion : The authors observed that Uncariae Ramulus et Uncus significantly down-regulates the expression of CD81, GFAP and MAG. These results suggest that Uncariae Ramulus et Uncus can be a candidate to regenerate CNS injury.

• 한국동물생명공학회지
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• 제38권4호
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• pp.189-198
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• 2023

Background: Brassica oleracea var. italica (broccoli), a rich source of antioxidants, can prevent various diseases and improve human health. In this study, we investigated the antioxidative effects of broccoli sprout extract on oxidative stress induced by lipopolysaccharide and cisplatin in cell and organ tissue models. Methods: Antioxidative effect of BSE was evaluated using DPPH and ABTS in RAW 364.7 cells, and effects of BSE on testes were investigated using Cisplatin-induced testicular damage model with an in vitro organ culture system. Results: The DPPH assay showed that the antioxidant activity of the alcoholic broccoli sprout extract was higher than that of the water extract. Additionally, the expression levels of antioxidation-related genes, Nrf2, Gsr, HO-1, and catalase, were significantly increased in broccoli sprout extract-treated RAW 264.7 cells, and the extract suppressed lipopolysaccharide-induced mitochondrial dysfunction. Based on the results in the RAW 264.7 cell culture, the antioxidative effects of the extracts were investigated in a mouse testis fragment culture. The expression of Nrf2, HO-1, and Ddx4 was clearly decreased in cisplatin-treated mouse testis fragments and not in both broccoli sprout extract- and cisplatin-treated mouse testis fragments. In addition, the oxidative marker O-HdG was strongly detected in cisplatin-treated mouse testis fragments, and these signals were reduced by broccoli sprout extract treatment. Conclusions: The results of this study show that broccoli sprout extracts could serve as potential nutraceutical agents as they possess antioxidant effects in the testes.

• 대한기계학회논문집A
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• 제39권11호
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• pp.1153-1159
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• 2015

최근 조직 공학 분야에서는 폴리카프로락톤(PCL), 수산화인회석, 삼인산칼슘, 이상인산칼슘(BCP), 지르코니아(Zirconia, $ZrO_2$) 와 같은 합성 생체폴리머와 생체세라믹 등은 다양한 생체 조직 또는 장기를 재생하는데 필요한 대체재로 사용되고 있다. 따라서, 본 연구의 목적은 골 조직 재생을 위한 혼합된 $ZrO_2$/BCP/PCL(ZBP) 인공지지체의 특성을 관찰하기 위함이다. 단선 패터닝 실험의 결과를 토대로 내부연결성 있는 공극을 가지고 $45^{\circ}+135^{\circ}$ 타입과 격자타입의 새로운 복합 공극 패턴을 가지는 혼합된 ZBP 인공지지체는 폴리머 적층시스템에 의해 성공적으로 제작되었다. 뿐만 아니라 기계적 특성에 대한 ZBP 인공지지체의 효과를 분석하였다. 게다가 MG63 세포에 대한 ZBP 인공지지체의 세포 상호작용은 CCK-8 분석을 이용함으로써 평가되었다.

• Molecules and Cells
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• 제26권1호
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• pp.93-99
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• 2008

Transcriptional silencing is regulated by promoter methylation and histone modifications such as methylation and acetylation. We constructed a Schizosaccaromyces pombe reporter strain, KCT120a, to identify modifiers of transcriptional silencing, by inserting the $ura4^+$ gene into a heterochromatic telomere region. Two compounds inhibited the activity of histone deacetylases, induced acetylation of histone H3 and caused apoptotic cell death in HeLa cells. Expression of gelsolin and $p21^{waf1/cip1}$ also increased, as it does in response to HDAC inhibitors such as TSA. Therefore, these compounds appear to be potent inhibitors of HDACs, and hence potential anti-cancer drugs. Our observations suggest that a yeast cell-based assay system for transcriptional silencing may be useful for identifying histone deacetylase inhibitors and other agents affecting chromatin remodeling.

• KSBB Journal
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• 제21권2호
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• pp.85-89
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• 2006

F. trogii ATCC 200800으로부터 원형질체 분리를 통해 단일균주들을 선별하였으며, 선별된 균주들의 고체배양 및 agar plug assay를 통해 효소생산을 위한 균주를 대량 선별하였다. Agar plug assay를 통해 4일 동안 100여종 이상의 균주를 동시에 배양, 분석이 가능하였으며, 염료분해환을 형성하지 않은 균주는 액체배양 확인 결과 MnP의 생산이 거의 일어나지 않는 것으로 나타났다. 이러한 방법으로 선별된 균주를 이용하여 UV 돌연변이를 통해 모균주로부터 유전적 변이를 유도해 새로운 균주선별을 시도하였으며, 이로부터 모균주와 비교하여 효소생산성이 향상되고 안정성이 증대된 균주를 선별할 수 있었다. 모균주와 선별된 균주의 명확한 유전적 차이를 규명하기보다는 배양시 형태학적 특성이 상이함을 확인하였다. 또한, 선별된 균주를 이용하여 배양액내에 다양한 종류의 inducer를 첨가에 따라 효소생산에 미치는 영향을 확인하였다. 과량의 inducer 물질이 첨가될 경우, 균체성장은 물론 효소생산성도 크게 떨어졌으며, 균체성장이 어느 정도 이루어진 상태 즉, 이차대사가 이루어지는 시점에서 inducer를 첨가할 경우 그 효율이 최대였으며, 동시에 inducer를 첨가할 경우 보다 미량씩 일정량을 나누어 주입할 경우 효소생산에 더 유리한 것으로 나타났다.

• Journal of Microbiology and Biotechnology
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• 제31권12호
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• pp.1716-1721
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• 2021

Chikungunya fever is an arboviral disease caused by the Chikungunya virus (CHIKV). The disease has similar clinical manifestations with other acute febrile illnesses which complicates differential diagnosis in low-resource settings. We aimed to develop a rapid test for CHIKV detection based on the nucleic acid lateral flow immunoassay technology. The system consists of a primer set that recognizes the E1 region of the CHIKV genome and test strips in an enclosed cassette which are used to detect amplicons labeled with FITC/biotin. Amplification of the viral genome was done using open-source PCR, a low-cost open-source thermal cycler. Assay performance was evaluated using a panel of RNA isolated from patients' blood with confirmed CHIKV (n = 8) and dengue virus (n = 20) infection. The open-source PCR-NALFIA platform had a limit of detection of 10 RNA copies/ml. The assay had a sensitivity and specificity of 100% (95% CI: 67.56% - 100%) and 100% (95% CI: 83.89% - 100%), respectively, compared to reference standards of any positive virus culture on C6/36 cell lines and/or qRT-PCR. Further evaluation of its performance using a larger sample size may provide important data to extend its usefulness, especially its utilization in the peripheral healthcare facilities with scarce resources and outbreak situations.

• 한국식품위생안전성학회지
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• 제33권1호
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• pp.50-57
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• 2018

Salmonella는 전세계적으로 식중독을 유발하는 주요 원인 균으로서, 식중독을 유발하는 Salmonella를 신속하게 검출하는 방법은 식품 안전을 위한 중요한 도구이다. Real-time PCR은 식중독균을 검출하기 위한 신속검사법으로 널리 사용되어 왔다. 최근에는 NBS LabChip real-time PCR이라는 새로운 시스템이 칩타입으로 조작이 간편하며 초고속의 real-time PCR 시스템이라는 보고가 있었다. 본 연구에서는 살모넬라의 신속한 검출을 위하여 NBS LabChip real-time PCR에 기반하여 real-time PCR 반응 시간이 20분 이내인 검출법을 확인하고자 하였다. 프라이머와 프로브 설계를 위해 두 개의 타겟 유전자(invA, stn)가 선택되었으며, 특이도와 민감도(검출한계)를 평가함으로 개발된 검출법을 검증하고자 하였다. 본 연구에서는 특이도 검증을 위해 Salmonella 균주 42주와 Non-Salmonella 균주 21주를 포함하였으며, 본 방법으로 Salmonella 42주에 대해서만 정확하게 검출이 가능하였다. 검출한계는 살모넬라 genome DNA 기준으로 $10^1copies/{\mu}L$으며, 소시지에서는 4시간 증균 이후 접종균수로서 $10^1CFU/g$에서 $10^2CFU/g$까지 검출이 가능하였다. 본 연구에서 개발된 검출법은 신속한 식중독 원인조사에 활용될 수 있을 것으로 기대된다.

• 동의생리병리학회지
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• 제24권4호
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• pp.586-591
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• 2010

Chemokine and Growth Factor are major mediumtors of immuno-inflammatory pathway. The purpose of this study is to investigate whether productions of Chemokine and Growth Factor in lipopolysaccharide (LPS)-induced mouse macrophage RAW 264.7 cells are modulated by Gallic acid (GA), which is easily founded in tannin-containing natural materials such as red wine, green tea, grape juice, and Corni Fructus. Productions of Chemokine and Growth Factor were analyzed by High-throughput Multiplex Bead based Assay with Bio-plex Suspension Array System based on $xMAP^{(R)}$ (multi-analyte profiling beads) technology. At first, cell culture supernatant was obtained after treatment with LPS and GA for 24 hour. Then, the antibody-conjugated beads were added and incubated for 30 minutes. After incubation, detection antibody was added and incubated for 30 minutes. And Strepavidin-conjugated Phycoerythrin (SAPE) was added. After incubation for 30 minutes, the level of SAPE fluorescence was analyzed on Bio-plex Suspension Array System. Based on fluorescence intensity, concentrations of Chemokine and Growth Factor were determined. The results of the experiment are as follows. GA significantly inhibited the production of interferon-inducible protein (IP)-10, keratinocyte-derived chemokine(KC), and vascular endothelial growth factor (VEGF) in LPS-induced RAW 264.7 cells at the concentration of 25, 50, 100, 200 uM (p<0.05). GA significantly inhibited the production of monocyte chemoattractant protein-1(MCP-1) and macrophage-colony stimulating factor(M-CSF) in LPS-induced RAW 264.7 cells at the concentration of 50, 100, 200 uM (p<0.05). GA diminished the production of granulocyte macrophage-colony stimulating factor (GM-CSF) in LPS-induced RAW 264.7 cells. But GA did not show the inhibitory effect on the production of leukemia inhibitory factor (LIP) and macrophage inflammatory protein (MIP)-2 in LPS-induced RAW 264.7 cells. These results suggest that GA has the immuno-modulating activity related with its inhibitory effects on the production of IP-10, KC, MCP-1, VEGF, and M-CSF in LPS-induced macrophages.

• 한국산학기술학회논문지
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• 제16권11호
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• pp.7575-7581
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• 2015

corticotropin releasing factor(CRF)는 스트레스에 의해 유도되는 신경펩타이드 물질들 중 하나로서 모발의 손실 및 재성장에 영향을 미친다고 광범위하게 제기되어 왔다. 이에 CRF1 수용체 길항제 개발을 위하여 세포 내 칼슘 신호전달 기전을 이용한 스크리닝 시스템을 개발하고 최적화 연구를 수행하고자 하였다. 이를 위하여 에쿼린 모체세포에 CRF1 수용체와 만능 G 단백질인 G${\alpha}$16 유전자를 동시에 발현시켜 안정화 세포주를 구축하였다(HEK293a16/hCRF1). 표준 효현제인 sauvagine의 반응이 임시 발현세포와 비교하여 안정화 세포주에서 농도 의존적 반응 범위가 12배 이상 증가하였으며($EC_{50}:15.21{\pm}1.83nM$), 이에 따라 길항제 스크리닝에 필수적인 안정적인 신호와 높은 용매 허용도를 확보할 수 있었다. CRF1 수용체에 대한 표준 길항제인 antalarmin과 CP154526에 대한 $IC_{50}$ 수치는 각각 $414.1{\pm}5.5$와 $290.7{\pm}1.9nM$로 확인되었는데 냉동보관세포의 경우에도 유사한 결과를 얻었다. 에쿼린 기반 세포 기능실험의 최적화 연구를 통해 구축된 CRF 수용체 안정화 세포주는 모발의 재형성과 관련된 신규의 기능성 화장품 및 조절물질 개발 연구에 적극적으로 활용 가능 할 것으로 판단된다.