• Macromolecular Research
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• v.17 no.4
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• pp.232-239
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• 2009

This paper describes the simple nanopatterning of proteins on polyelectrolyte surfaces using microcontact printing with a nanopatternable, hydrophilic composite nanomold. The composite nanomold was easily fabricated by blending two UV-curable materials composed of Norland Optical Adhesives(NOA) 63 and poly(ethylene glycol) dimethacrylate(PEG-DMA). NOA 63 provided stable nanostructure formation and PEG-DMA induced high wettability of proteins in the nanomold. Using the composite mold and functionalized surface with polyelectrolytes, the fluorescent, isothiocyanate-tagged, bovine serum albumin(FITC-BSA) was successfully patterned with 8 nm height and 500 nm width. To confirm the feasibility of the protein assay on a nanoscale, a glycoprotein-lectin assay was successfully demonstrated as a model system. As expected, the lectins correctly recognized the nano-patterned glycoproteins such as chicken ovalbumin. The simple preparation of composite nanomold and functionalized surface with a universal platform can be applied to various biomolecules such as DNA, proteins, carbohydrates, and other biomolecules on a nanoscale.

• Journal of the korean society of oceanography
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• v.35 no.3
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• pp.153-157
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• 2000

Four different FITC-conjugated lectins were used to visually evaluate lectin binding activity by optical staining quality using confocal laser scanning microscopy (CLSM) of Cochzodinium polykrikoides in nature (wild type) and culture (cultured type). Cells from the field and cultures treated with ConA fluoresced only at the outer cell wall, and the abundance and distribution of the fluorescent signal were similar. Treatment with PWM and HPA did not elicit fluorescence at the cell surface, but the wild type exposed to HPA showed greater binding than did the cultured cells, possibly due to greater concentrations of glucosamine. The wild type cells treated with LBL lectin showed a strong green fluorescence on the cell surface, whereas cultured cells did not. Signal intensity and abundance were greater than for any other lectins tested in this study. These results suggest that wild type and cultured type are significantly different based on surface sugar production. In particular, the wild type cells apear richer in galactosamine-like moieties. Neither glucose nor mannose-like moieties were present in either wild types or cultured cells.

• Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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• 2001.07a
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• pp.757-760
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• 2001

We have used the random fluidic self-assembly (RFSA) technique based on the chip pattern of hydrophobic self-assembly layers to assemble microfabricated particles onto the chip pattern. Immobilization of DNA, fabrication of the particles and the chip pattern, arrangement of the particles on the chip pattern, and recognition of each using DNA fluorescence measurement were carried out. Establishing the walls, the arrangement stability of the particles was improved. Each DNA is able to distinguish by using the lithography process on the particles. Advantages of this method are process simplicity, wide applicability and stability. It is thought that this method can be applicable as a new fabrication technology to develop a minute integration type biosensor microarray.

• Macromolecular Research
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• v.15 no.6
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• pp.513-519
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• 2007

Self-assembled nanoparticles have great potential to act as vehicles for hydrophobic drug delivery. Understanding nanoparticle cellular internalization is essential for designing drugs intended for intracellular delivery. Here, the endocytosis and exocytosis of fluorescein isothiocyanate (FITC)-conjugated glycol chitosan (FGC) self-assembled nanoparticles were investigated by flow cytometry and confocal microscopy. The cellular internalization of FGC nanoparticles was initiated by nonspecific interactions between nanoparticles and cell membranes. Although adsorptive endocytosis of the nanoparticles occurred quickly, significant amounts of FGC nanoparticles were exocytosed, particularly in the early stage of endocytosis. The amount of exocytosed nanoparticles was dependent on the pre-incubation time with nanoparticles, suggesting that exocytosis is dependent on the progress of endocytosis. FGC nanoparticles internalized by adsorptive endocytosis were distributed in the cytoplasm, but not in the nucleus. In vitro cell cycle analysis demonstrated that FGC nanoparticles delivered paclitaxel into the cytoplasm and were effective in arresting cancer cell growth.

• Biomolecules & Therapeutics
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• v.8 no.1
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• pp.22-26
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• 2000

In order to study the cytotoxic properties of sesquitepenes, dehydrocostus lactone (DL) and costunolide from Saussurea lappa, cytotoxicity was measured by SRB method using various human cancer cell lines. Dehydrocostus lactone(DL) and costunolide exhibited significant cytotoxicity against A-549, SK-OV-3, SK-MEL-2, XF-498 and HCT 15 cells. The U937 human leukemia cells treated with DL showed several apoptotic evidences like chromosome condensation and formation of apoptotic bodies. From the results of FACS analysis, early apoptosis was observed by phosphatidylserine externalization detected by annexin V-FITC. Furethermore, these studies determined hypodiploid contents and effects on the cell phase distribution of DL-treated U937 cells. After exposure of U937 cells to $30\mu\textrm{M}$ DL effectively led to G2/M modified cell cycle distribution within 24hr. These observations suggest that DL can be used efficiently for the cancer treatment.

• Proceedings of the KSMRM Conference
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• 2003.10a
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• pp.36-36
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• 2003

목적： 고형성 종양에 특이적으로 발현하는 MR 분자영상용 조영제의 후보물질을 분자생물학적 방법 및 in vitro 분자영상기법을 이용하여 탐색하고자 하였다. 대상 및 방법： 암이 진행됨에 따라 유전자가 과발현되는 세포표면 단백질인 CD44 와 세포막에 존재하는 단백질분해효소의 일종인 MTI-MMP를 일차 선정하여 유전자 수준 및 단백질 수준에서 분자생물학적 방법으로 종양특이성에 대해 검증하였고, 특히 각종 암세포에서 발현양이 증가하는 CD44의 변형인 CD44v6 항체를 이용하여 생세포에서의 표적화에 대한 광학적 검증을 위해 초상자성 나노입자 대신 형광입자(FITC)를 결합하여 confocal microscopy로 실시간 표적화에 대한 분자영상을 획득하였다.

• Fisheries and Aquatic Sciences
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• v.3 no.2
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• pp.83-87
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• 2000

We have investigated lectin binding patterns in order to apply binding records of previous laboratory experiments to field settings before the first ourbreaks of harmful algal bloom (HAB). Although cells were grown under different conditions, the binding patterns were the same as in the control. In addition, culture days was not associated with the binding patterns, when compared with the control. In nature, this results suggest that ECA, HPA and WGA lectin are able to discriminate between C. polykrikoides and G. impudicum, as well as ECA and SBA have a capability as a tool for differentiating between C. polyrikoides and G. catenatum, although these species are closely similar under the light microscope fiexed with Lugol solution.

• Proceedings of the Korea Society of Poultry Science Conference
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• 2001.11a
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• pp.69-70
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• 2001

Lectins have great potential as to determine the alternation of the distribution of cell surface carbohydrates during cellular development and differentiation. Here, we investigated the presence and distribution of cell surface carbohydrates on chicken primordial germ cells (PGCs) during the migration and gonadal stages using a variety of lectins. A total of six FITC-labelled lectins from several specificity classes were used: ConA (glucose/mannose), WGA (N-acetylglucosamine), STA (N-acetylglucosamine), DBA (N-acetylgalactosamine/galactose), UEA-I (fucose) and PHA-E (oilgosaccharide). As a results, PGC-specific binding was observed in STA. PGCs of migration stage (2.5- and 5.5-day embyos) were STA-positive whereas PGCs of 10-day embryonic gonad were not. The results suggest that N-acetylglucosamine residuse are present specifically in migrating chicken PGCs and changes during development.

• Journal of the Korean Society of Visualization
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• v.11 no.1
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• pp.16-21
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• 2013

In this study, diffusion coefficients of 20 kDa FITC-dextran in 2% agarose gel with different cell concentrations were measured using fiberoptic-based fluorescence recovery after photobleaching technique. As increasing cell concentration suspended in agarose gel, the diffusion coefficients were decreased. The diffusion coefficient of agarose gel which contains $10{\times}10^6$ cells/ml was decreased to 11% that of in agarose gel without cells. The distribution of fluorescence dye in 3D scaffold was also simulated. The simulation result shows that the diffusion coefficient is more significant factor than the scaffold structure.

• Macromolecular Research
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• v.17 no.7
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• pp.464-468
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• 2009

Gelatin nanoparticles derived from bovine or porcine have been developed as various types of drug delivery system, and they need to be cross-linked to maintain their physicochemical properties in aqueous environments. Although gelatin is a widely used material in pharmaceutical industries, the safety issue of animal-origin gelatins, such as transmissible mad cow disease and anaphylaxis, remains to be solved. The purpose of this study was to prepare type A gelatin (GA) nanoparticles by modified, two-step, desolvation method and compare the toxicity of the resulting GA nanoparticles with recombinant human gelatin (rHG) nanoparticles. The GA nanoparticles were characterized, and drug loading and release pattern were measured. FITC-BSA, a model protein, was efficiently loaded in the nanoparticles and then released in a biphasic and sustained release pattern without an initial burst. In particular, the cell viability of the GA nanoparticles was less than that of the rHG nanoparticles. This finding suggests that rHG nanoparticles should be considered as an alternative to animal-origin gelatin nanoparticles in order to minimize the safety problems.