• Journal of the Korean Society of Manufacturing Technology Engineers
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• v.26 no.1
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• pp.1-5
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• 2017

This study investigated the mechanical tearing of a cell membrane using a nanostructured alumina filter for easy and quick mechanical cell disruption. Nanostructured alumina filters were prepared by a multi-step aluminum anodizing process and nanopore etching process. Six different types of nanostructures were formed on the surface of the nanoporous alumina filters to compare the mechanical cell disruption characteristics according to the shape of the nanostructure. The prepared alumina filter was assembled in a commercial filter holder, and then, NIH3T3 fibroblast cells in a buffer solution were passed through the nanostructured alumina filter at a constant pressure. By measuring the concentration of proteins and DNA, the characteristics of mechanical cell disruption of the nanostructured alumina filter were investigated.

• Journal of the Korean Society of Manufacturing Process Engineers
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• v.20 no.8
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• pp.33-41
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• 2021

Recently, many studies have been conducted on substituting fossil fuels with bio-refineries in existing industrial systems using biomass. Among the various bio-refineries, microalgae have received wide attention because it uses inorganic compounds to produce useful substances, which are extracted by a cell disruption process. Although numerous cell disruption methods exist, cell disruption efficiency has been studied by ultrasonic treatment. Ultrasound is a high-frequency (20 kHz or higher) sound wave and causes cell disruption by cavitation when passing through a solvent. In this study, we used the microalgal species Chlorella sp., which was cultured in a plate-type photobioreactor. The experiment was conducted using a continuous low-frequency processing device. The reduction of cells with time due to cell disruption was fitted using a logistic model, and optimum conditions for highly efficient cell disruption were determined by conducting experiments under multiple conditions.

• Journal of the Korean Society of Manufacturing Process Engineers
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• v.19 no.2
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• pp.111-118
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• 2020

Recently, bioenergy research using microalgae, one of the most promising biofuel sources, has attracted much attention. Cell disruption, which can be classified as physical or chemical, is essential to extract functional ingredients from microalgae. In this study, we investigated the cell disruption efficiency of Chlorella sp. using low-frequency non-focused ultrasound (LFNFU). This is a continuously physical method that is superior to chemical methods with respect to environmental friendliness and low processing cost. A flat panel photobioreactor was employed to cultivate Chlorella sp. and its growth curve was fitted both with Logistic and Gompertz models. The temporal change in cell reduction by cell disruption using LFNFU was fitted with a Logistic model. The experimental conditions that were investigated were the initial concentration of microalgal cells, relative amplitude of output ultrasound waves, processing volume of microalgal cells, and initial pH value. The optimal conditions for the most efficient cell disruption were determined through the various tests.

• Journal of the Korean Society of Manufacturing Process Engineers
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• v.20 no.10
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• pp.63-71
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• 2021

Using fossil fuels in existing industrial systems causes a variety of social problems. Recently, many studies have been conducted on bio-refineries, which aim to actively utilize biomass to reduce the use of fossil fuels and solve various social problems. Among them, research using microalgae as a third-generation biomass has attracted considerable attention. Microalgae use inorganic matter to produce organic matter, and cell destruction is necessary to extract useful organic materials from microalgae. The extracted organic materials are currently used in various industrial fields. Numerous cell-destruction methods exist. We have investigated cell disruption by sonication, especially its efficiency. Ultrasound is a sound wave with frequencies above 20 kHz, and destroys cells by sending high energy through a cavitation that occurs, according to the characteristics of the sound wave. The Dunaliella salina microalgae used in this study was cultured in a flat-type photobioreactor. Experiments were performed using a batch low-frequency processing device. Logistic model was applied to analyze the results of cell-destruction experiments using ultrasound. The proper conditions for the most efficient cell destruction were OD 1.4(microalgae concentration)), 54watt(output power) and 200mL(microalgae capacity).

• Transactions of the Korean Society of Mechanical Engineers A
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• v.32 no.10
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• pp.831-835
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• 2008

We present a continuous electrical cell lysis chip, using a DC bias voltage to generate the focused high electric field for cell lysis as well as the electroosmotic flow for cell transport. The previous cell lysis chips apply an AC voltage between micro-gap electrodes for cell lysis and use pumps or valves for cell transport. The present DC chip generates high electrical field by reducing the width of the channel between a DC electrode pair, while the previous AC chips reducing the gap between an AC electrode pair. The present chip performs continuous cell pumping without using additional flow source, while the previous chips need additional pumps or valves for the discontinuous cell loading and unloading in the lysis chambers. The experimental study features an orifice whose width and length is 20 times narrower and 175 times shorter than the width and length of a microchannel. With an operational voltage of 50 V, the present chip generates high electric field strength of 1.2 kV/cm at the orifice to disrupt cells with 100% lysis rate of Red Blood Cells and low electric field strength of 60 V/cm at the microchannel to generate an electroosmotic flow of $30{\mu}m/s{\pm}9{\mu}m/s$. In conclusion, the present chip is capable of continuous self-pumping cell lysis at a low voltage; thus, it is suitable for a sample pretreatment component of a micro total analysis system or lab-on-a-chip.

• Toxicological Research
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• v.11 no.1
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• pp.103-108
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• 1995

Rat renal proximal tubule suspension was prepared from adult male Sprague Dawley rat (250-300g) by mechanical (non-enzymatical) method and evaluated as a pontential model for mechanistic studies and early screening of nephrotoxicity, using anionic antibiotics (cephaloridine). Cephaloridine (CPL) produced an increase in LDH release into media. This release results from decrease a proximal tubule cell viability and subsequently increase the permeability of cell viability and subsequently increase the permeability of cell membrane. Since loss of intracellular potassium and ATP into media is the sign of disruption of cell membrane, especially basolateral membrane (BLM), CPL induced proximal tubule cell compromise also appear be associated with BLM, maybe $Na^+-K^+$ ATPase. Also seen was significant depression in brush border membrane (BBM) ALP activity and no significantly increase in BBM GGT activities. The inhibition of typical anion, PAH accumulation (especially, CPL 5 mM) and cation, TEA (especially, 4hours incubation) were seen dose dependently. This is because of CPL accumulation in renal proximal tubule and increase of cytotoxicity.

• Applied Microscopy
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• v.27 no.2
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• pp.121-130
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• 1997

It has been demonstrated that majority of cells in the mammalian body such as myocytes and epithelial cells of skin and intestine respond to mechanical force or environmental factors and exhibit partial disruption of cell membrane, i. e., cell wounding, even in a physiological condition. Myocardial cells are rather apt to be wounded than other cells since they are definitely exposed to mechanical stress by contraction-relaxation and blood flow. However, the mechanism how myocardial cells protect themselves against cell wounding is not yet clarified. On this background, the present study was performed to elucidate whether albumin leakage is related to cell wounding and to assess whether diltiazem, a potent calcium channel blocker, is beneficial in isoproterenol-induced cell wounding in the heart. Hearts isolated from New Zealand White rabbits ($1.5\sim2.0kg$ body weight, n=20) were perfused with Tyrode solution by Langendorff technique. After stabilization of baseline hemodynamics, the hearts were subjected to bolus administration of isoproterenol and diltiazem as following order: $1.6{\mu}M$ isoproterenol at zero min (the beginning point): $16{\mu}M$ diltiazem at 20min; $1.6{\mu}M$ isoproterenol at 25min; $16{\mu}M$ isoproterenol at 45 min; $160{\mu}M$ diltiazem at 65 min; $16{\mu}M$ isoproterenol at 70 min. During all experiments, the left ventricular function was recorded, albumin leakage in the coronary effluents was analyzed by electrophoresis and Western blot, and myocardial cell membranes were examined by conventional transmission electron microscopy. Data were analyzed by t-test and linear regression test. Isoproterenol significantly increased the inotropic and chronotropic contractions, coronary flow, and frequency of arrhythmia, however, diltiazem did not influence on hemodynamics except decrease in the frequency of arrhythmia and a slight decrease in contractility. Isoproterenol also resulted partial disruption of myocardial cell membrane and inclose in albumin leakage, while diltiazem pretreatment showed number of electron-dense plaques in the cell membrane and a tendency of decrease in albumin leakage. These results indicate that albumin leakage may be an indirect index of cell wounding in the heart and diltiazem nay be beneficial to protect myocardial cells against isoproterenol-induced cell wounding. It is likely that diltiazem promotes resealing process of the cell membrane.

• Proceedings of the KOSOMBE Conference
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• v.1994 no.12
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• pp.181-184
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• 1994

• Proceedings of the KSME Conference
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• 2004.11a
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• pp.545-550
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• 2004

KSTAR cryostat is a 8.8 m diameter vacuum vessel that provides the necessary thermal barrier between the ambient temperature test cell and the supercritical helium cooled superconducting magnet providing the base pressure of 1 ${\times}$ $10^{-3}Pa$. The cryostat is a single walled vessel consisting of central cylindrical section and two end closures, a flat base structure with external reinforcements and a dome-shaped lid structure. The base structure has 8 equally spaced support legs anchored on the concrete base. The cryostat vessel design was executed to satisfy the performance and operation requirements. The major loads considered in the structural analysis were vacuum pressure, dead weight, electromagnetic load driven by plasma disruption, and seismic load. Based on the fabrication and inspection procedures for the vessel, cryostat vessel was fabricated and inspected. It was confirmed that the inspection results were acceptable.

• KSBB Journal
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• v.30 no.6
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• pp.291-295
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• 2015

The limited productivity of natural shell matrix proteins has hampered the investigation of their biochemical properties and practical applications, although biominerals in nature obtained by organic-inorganic assemblies have attractive mechanical and biological properties. Here, we prepared a vector for the expression of a fusion protein of a shell matrix protein from Pinctada fucata (named as GRP_BA) with the GRGDSP residue. The fusion protein of BA-RGD was simply produced in E. coli and purified through sequential steps including the treatment with $CaCl_2$ and EDTA solution for cell membrane washing, mechanical cell disruption and the application of non-ionic surfactant of Triton X-100 for BA-RGD inclusion body washing. The production yield was approximately 60 mg/L, any other protein band was not observed in SDS-PAGE and it was estimated that above 97% endotoxin was removed compared to the endotoxin level of whole cell. This study showed this simple and easy purification approach could be applied to the purification of BA-RGD fusion protein. It is expected that the protein could be utilized for the preparation of biominerals in practical aspects.