• KSBB Journal
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• v.15 no.1
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• pp.35-41
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• 2000

We tried to prepare encapsulated whole cell cyclodextrin glucanotransferase(CGTase) in order to produce glycosyl-xylitol using xylitol as glucosyl acceptor. The organic nitrogen source was more effective for the production of CGTase from Bacillus macerans IFO 3490 than the inorganic one. Most of the CGTase which had been produced during cultivation was excreted to the growth medium. B. macerans cells inocculated in the capsule failed to grow to the high cell density. Adsorbents such as activated charcoal, Sephadex and Amberite resins could not adsorb efficiently the CGTase from the broth solution. We obtained successfully the encapsulated whole cell CGTase by immobilizing the concentrated broth solution in the calcium alginate capsules. The encapsulated whole cell CGTase carried out the transglycosylation reaction which converts xylitol into glucosyl-xylitol using dextrin as glucosyl donor.

• Journal of Embryo Transfer
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• v.31 no.1
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• pp.81-88
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• 2016

There are various factors i.e. donor cell type, culture system as well as technical procedures which influence the pre-implantation embryonic development; however, may attempts have been made and still it is under investigation to improve the cloning efficiency using somatic cell nuclear transfer technique. It is has been investigated that stem cells like mesenchymal stem cell are able to more efficiently reprogram and reactivate the expression of early embryonic genes to promote nuclear transfer efficiency. In addition, Oct4 expression plays a pivotal role in early embryo development. In the present study, we investigated distribution of Oct4 expression and changes of apoptosis and total cell number in nuclear transfer blastocyst after using Oct4 transfected bone marrow stem cell as donor cells. Although Oct4-RFP expression was observed across blastocyst, more concentrated intensity was shown at hatched region in blastocyst on day 7. Reduction of apoptotic bodies was revealed in Oct4 transfected blastocyst by TUNEL staining, however, there was no significant difference in total cell number between Oct4 transfected and non-transfected nuclear transfer embryos. In conclusion, Oct4 transfected donor cells exhibited higher expression in hatching sight in day 7 blastocyst and were able to prevent apoptosis compared to non-transfected donor cells.

• Korean Journal of Clinical Laboratory Science
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• v.45 no.1
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• pp.37-42
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• 2013

The cells are concentrated approximately 20-fold by cytocentrifugation. This study evaluated the nucleated cell number for cells recovered on slide by using Cytospin-3 (Thermo Shandon Ltd. UK) and Cytopro-7620 (Wescor Inc., USA) cytocentrifuges to hematocytometer cell count of $0{\sim}5WBCs/{\mu}L$ of hematocytometer in the cerebrospinal fluid cell count. One hundred forty eight samples of $0{\sim}5WBCs/{\mu}L$ on hematocytometer, were cytocentrifuged by Cytospin-3 and Cytopro-7620 instruments. The nucleated cell number for cells recovered on slide was counted after Wright stain. The nucleated cell number for cells recovered on slide was 0~40 cells in the 44 samples of $0WBC/{\mu}L$, and 3~95 cells in the 31 samples of $1WBC/{\mu}L$. It was observed that the nucleated cell number for cells recovered on slide was 13~100 cells in the 44 samples of $2WBCs/{\mu}L$, and more than 100 cells in the 29 samples of $3{\sim}5WBCs/{\mu}L$, respectively. In addition, extremely normal lymphocyte, monocyte and polymorphonuclear neutrophil were observed in the 143 samples of $0{\sim}5WBCs/{\mu}L$. Macrophage and eosinophil were also rarely observed. The nucleated cell number for cells recovered on slide was 20 cells, which were regarded as $1WBC/{\mu}L$ in body fluid cell count. However, in this study, we made alterations to report nucleated cell percentage as 0% without preparing the cytocentrifuged slide at $0WBC/{\mu}L$ by using the cell yield in a comparison between the value of $0{\sim}5WBCs/{\mu}L$ and nucleated cell number for cells recovered on slide.

• Biotechnology and Bioprocess Engineering:BBE
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• v.10 no.3
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• pp.284-288
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• 2005

Baker's yeast was disrupted in a 1.4-L stainless steel horizontal bead mill under a continuous recycle mode using 0.3 mm diameter zirconia beads as abrasive. A single pass in continuous mode bead mill operation liberates half of the maximally released protein. The maximum total protein release can only be achieved after passaging the cells 5 times through the disruption chamber. The degree of cell disruption was increased with the increase in feeding rate, but the total protein release was highest at the middle range of feeding rate (45 L/h). The total protein release was increased with an increase in biomass concentration from 10 to $50\%$(w/v). However, higher heat dissipation as a result of high viscosity of concentrated biomass led to the denaturation of labile protein such as glucose 6-phosphate dehydrogenase (G6PDH). As a result the highest specific activity of G6PDH was achieved at biomass concentration of $20\%$(ww/v). Generally, the degree of cell disruption and total protein released were increased with an increase in impeller tip speed, but the specific activity of G6PDH was decreased substantially at higher impeller tip speed (14 m/s). Both the degree of cell disruption and total protein release increased, as the bead loading increased from 75 to $85\% (v/v)$. Hence, in order to obtain a higher yield of labile protein such as G6PDH, the yeast cell should not be disrupted at biomass concentration and impeller tip speed higher than $20\%(w/v)$ and 10 m/s, respectively.

• KSBB Journal
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• v.15 no.3
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• pp.219-225
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• 2000

Both the production of high spore concentration and high insecticidal activity are required in the production of Bacillus thuringiensis to be used for the bacterial insecticide. In the production of high cell and spore concentrations of B. thuringiensis the continuous fed-batch culture(CFBC) and intermittent fed-batch culture(IFBC) were investigated at $28^{\circ}C$ by maintaining 40% dissolved oxygen concentration. When the final glucose concentration was 50 g/L the maximum viable cell number obtained using the CFBC with linear gradient feeding was $9.37{\times}109$ cells/mL and maximum spore concentration was $8.33{\times}109$ spores/mL which was approximately 84.4% yield of spore formation. When the final glucose concentration was 100 g/L the aximum viable cell and spore concentrations obtained using the IFBC with pH-statb were $1.38{\times}$1010 cells/mL and $1.35{\times}1010$ spores/mL respectively and the yield of spore formation was approximately 97.8%.

• Transactions of the Korean Society of Automotive Engineers
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• v.21 no.1
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• pp.43-50
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• 2013

The stringent emission regulation and future shortage of fossil fuel motivate the research of alternative powertrain. In this study, a system of proton exchange membrane fuel cell has been modeled to analyze the performance of the fuel cell system for automotive application. The model is composed of the fuel cell stack, air compressor, humidifier, and intercooler, and hydrogen supply which are implemented by using the Matlab/Simulink(R). Fuel cell stack model is empirical model but the water transport model is included so that the system performance can be predicted over various humidity conditions. On the other hand, the model of air compressor is composed of motor, static air compressor, and some manifolds so that the motor dynamics and manifold dynamics can be investigated. Since the model is concentrated on the strategic operation of compressor to reduce the power consumption, other balance of components (BOP) are modeled to be static components. Since the air compressor model is empirical model which is based on curve fitting of experiments, the stack model is validated with the commercial software and the experiments. The dynamics of air compressor is investigated over unit change of system load. The results shows that the power consumption of air compressor is about 12% to 25% of stack gross power and dynamic response should be reduced to optimize the system operation.

• Applied Microscopy
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• v.35 no.3
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• pp.177-186
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• 2005

The foregut epithelium of the last instar larva in the scarab beetle, Allomyrina dichotoma was observed with electron microscopes. The foregut epithelium of the scarab beetle larva is composed of a single-layered squamous absorptive cell. The luminal surface of the epithelium is covered with cuticular intima. The free surface of the squamous cell has a irregular array of microvilli 'brush border', while cell membrans close to the basal lamina are infolded and a lot of mitochondria are concentrated in those processes. The cytoplasm in the epithelial cells is well developed nucleus, mitochondria. And the basal region of cell contains large lipid-, protein droplets and numerous glycogen granules. The basal lamina is located between the basal membrane and muscle bundle, providing barrier between the epithelium and the hemolymph. The epithelium is surround by the subepithelial space and muscles. The subepithelial space, which is composed of fibrous connective tissue is innervated by many tracheoles and axon.

• YAKHAK HOEJI
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• v.47 no.6
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• pp.410-416
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• 2003

Autotaxin(ATX) was originally purified from conditioned media of A2058 human melanoma cells and shown to be a potent cell motility-stimulating factor, possessing a type II nucleotide pyrophosphatase/phosphodiesterase (NPP2) activity. Recombinant ATX has recently demonstrated that human plasma lysophosholipase D is identical to ATX and uses lysophosphatidylcholine as a substrate to mediate various biological functions including tumor cell growth and motility through G-protein coupled receptor. However, despite pivotal roles of ATX on physiological or pathophysiological states, the production of ATX is solely depends on complicated purification method which employs multiple column steps, but resulted in very poor yield. This limited the use of ATX for extensive analysis. We, therefore, expressed six histidine-tagged recombinant human ATX(His-ATX) in High Five TM insect cells to improve the generation of ATX and to make simple the purification of ATX. The signal sequence of the human ATX gene was truncated and replaced with sequence of insect cell secretion signal within expression vector. In addition, codons for six histidines were added to the C-termini of 120kDa ATX cDNA construct. A simple purification scheme utilizing two-step affinity column chromatography was designed to purify His-ATX to homogeneity from the culture supernatant of transfected insect cells. Homogenous His-ATX was detected and isolated from the concentrated insect cell medium using concanavalin A agarose and nickel affinity chromatography. Purified His-ATX was in full length with ATX capacity. A combination of this expression system and purification scheme would be useful for production and purification of high-quality functional ATX for research and practical application of multiple functional motogen, ATX/NPP-2.

• Biomaterials Research
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• v.22 no.4
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• pp.352-359
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• 2018

Background: Black phosphorus (BP) has emerged as a novel class of nanomaterials owing to its unique optical and electronic properties. BP, a two-dimensional (2D) nanomaterial, is a structure where phosphorenes are stacked together in layers by van der Waals interactions. However, although BP nanodots have many advantages, their biosafety and biological effect have not yet been elucidated as compared to the other nanomaterials. Therefore, it is particularly important to assess the cytotoxicity of BP nanodots for exploring their potentials as novel biomaterials. Methods: BP nanodots were prepared by exfoliation with a modified ultrasonication-assisted solution method. The physicochemical properties of BP nanodots were characterized by transmission electron microscopy, dynamic light scattering, Raman spectroscopy, and X-ray diffractometry. In addition, the cytotoxicity of BP nanodots against C2C12 myoblasts was evaluated. Moreover, their cell imaging potential was investigated. Results: Herein, we concentrated on evaluating the cytotoxicity of BP nanodots and investigating their cell imaging potential. It was revealed that the BP nanodots were cytocompatible at a low concentration, although the cell viability was decreased with increasing BP nanodot concentration. Furthermore, our results demonstrated that the cells took up the BP nanodots, and the BP nanodots exhibited green fluorescence. Conclusions: In conclusion, our findings suggest that the BP nanodots have suitable biocompatibility, and are promising candidates as fluorescence probes for biomedical imaging applications.

• Korean Journal of Human Ecology
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• v.17 no.5
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• pp.1027-1035
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• 2008

Essentail oils and carrier oils are generally used for Aromatherapy. Therefore the toxicity, possibilities of irritations and sensitive reactions and injury of essential oils must be considered for clients and therapists. So that, in this studies a toxicity of jojoba and 4 species essential oils (fennel, mandarine, tea tree and cedarwood) were investigated by the measurement of MTT-assay and sirius red staining. Liver, kidney and brain tell were chosen for the cell viability assay and observation of morphological change. In the result, no cytotoxicity was observed on live., kidney and brain cell at concentration of 0.01 $\mu\el/m\el$ jojoba oil. And lysis and nucleus breaking were not observed at same concentration of jojoba oil on live., kidney and brain cell. fennel oil was showed 50% of cell viability and inhibited cell growth on liver, kidney and brain cell at relatively high concentration compared with the other oils. 50% of liver, kidney and brain cell viability and delayed cell growth of tea tree and mandarine oil were revealed at lower concentration than fennel oil. In cedarwood oil, 50% of liver cell viability at concentration of 0.00067 $\mu\el/m\el$ was showed, but cell viability and cell growth of kidney and beam cell were effected at the lowest concentration compared with other oils. So that, jojoba oil as using of carrier oil may be not harmful. And 3 essential oils from the fennel, tea tree and mamdarine may have very low toxicity, but cedarwood may be used carefully for inhalation. And over dosage of concentrated cedarwood oil should be not directly touched and exposured, and absolute essential oils must be diluted with carrier oils for topical and systematic massage.