• KSBB Journal
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• v.31 no.4
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• pp.270-276
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• 2016

In glycoprotein, Terminal sialic acid residues of N-linked glycan are imperative things because they prevent the recognition from asialoglycoprotein-receptor that affect the half-life of glycoproteins. So establishment of culture process for enhancing sialic acid is important to maximize sialic acid contents of glycoprotein. In this study, we investigated effects of biphasic culture of Chinese hamster ovary (CHO) cells producing albumin-erythropoietin to increase sialylation. Biphasic cultures were performed with shift of $CO_2$ concentrations and temperatures at day 5 when viable cell density was decreased and sialidase was started to be released by cell lysis. The examined temperature set points were 33, 35 and $37^{\circ}C$ respectively and the $CO_2$ concentration was 1, 5, 10 and 15%. We confirmed that sialidase activity was the lowest in biphasic culture that was shifted from normal culture condition to 1% of $CO_2$ and $33^{\circ}C$ on day 5. However, the temperature and concentration of $CO_2$ have little effect on activity of ${\alpha}2,3$-sialyltransferase. Also, sialic acid contents were enhanced 1.13-fold higher than that in control culture. In conclusion, Biphasic cultivation in CHO cells led to inhibition of sialidase activity and increases of sialylated glycan.

• 한국생물공학회:학술대회논문집
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• 2002.04a
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• pp.293-296
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• 2002

Hyperosmotic pressure increased specific antibody productivity ($q_{Ab}$) of recombinant CHO cells (SH2-0.32) while it depressed cell growth. Thus, the use of hyperosmolar medium did not increase the maximum antibody concentration substantially. To overcome this drawback, the feasibility of biphasic culture strategy was investigated. In the biphasic culture, cells were first cultivated in the standard medium with physiological osmolality(294 mOsm/kg) for cell growth. When cells reached the late exponential phase of growth, the spent standard medium was replaced with the fresh hyperosmolar medium (522 mOsm/kg) for antibody production. The ($q_{Ab}$) in growth phase with the standard medium was 2.1 ${\mu}g/10^6cell/day$ while the ($q_{Ab}$) in antibody production phase with the hyperosmolar medium (522 mOsm/kg) was 11.1 ${\mu}g/10^6cell/day$. Northern blot analysis showed a positive relationship between the relative contenet of Ig mRNA and ($q_{Ab}$), indicating that transcriptional regulation was involved in the response of rCHO cells to hyperosmotic pressure. Due to the enhanced ($q_{Ab}$) and increased cell concentration in biphasic culture, the maximum antibody concentration obtained in biphasic culture with 522 mOsm/kg medium exchange was 161% higher than that obtained in batch culture with the standard medium. Taken together, simple biphasic culture strategy based on hyperosmotic culture for improved foreign protein production from rCHO cells is effective in improving antibody production of rCHO cells.

• Tuberculosis and Respiratory Diseases
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• v.45 no.2
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• pp.416-420
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• 1998

Flow-volume loop is known to be useful in the diagnosis of upper airway obstruction. In cases of tracheal obstruction, characteristic features such as fixed or variable upper airway obstruction patterns give clue to the diagnosis. But the flow-volume loop of unilateral mainstem bronchial stenosis is not known well. There is controversy in patterns of flow-volume loop in unilateral mainstem bronchial stenosis (restrictive pattern or biphasic pattern). We report a case of biphasic flow-volume loop in left mainstem bronchial stenosis(4-5 mm in diameter) as a sequela of endobronchial tuberculosis, which recovered normal flow-volume loop after metallic stent insertion and 2 months later showed reappearance of biphasic pattern because of restenosis of left mainstem bronchus due to growth of granulation tissue at the stent site.

• Proceedings of the Korean Society of Precision Engineering Conference
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• 2012.10a
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• pp.161-162
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• 2012

• Proceedings of the Korean Ceranic Society Conference
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• 2002.10a
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• pp.203.1-203
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• 2002

• Bulletin of the Korean Chemical Society
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• v.32 no.6
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• pp.1945-1950
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• 2011

The pyridinolysis of methyl phenyl phosphinic chloride is investigated kinetically in acetonitrile at -20.0 $^{\circ}C$. The Hammett and Br${\o}$nsted plots for substituent X variations in the nucleophiles are biphasic concave downwards with a break point at X = H, and unusual positive ${\rho}_X$ (= 2.94) and negative ${\beta}_X$ (= -0.48) values are obtained for the strongly basic nucleophiles. A stepwise mechanism with a rate-limiting step change from bond breaking for the weakly basic pyridines to bond formation for the strongly basic pyridines is proposed on the basis of biphasic concave downward Hammett and Br${\o}$nsted plots. Unusual positive ${\rho}_X$ and negative ${\beta}_X$ values are rationalized by the isokinetic relationship. The pyridinolyses and anilinolyses of four $R_1R_2$P(=O)Cl-type substrates, dimethyl, diethyl, methyl phenyl, and diphenyl phosphinic chlorides in acetonitrile are compared to obtain systematic information on phosphoryl transfer reaction mechanism. The combination of the two ligands, Me and Ph, shows unexpected kinetic results for both the anilinolysis and pyridinolysis: greatest magnitude of $k_H/k_D$ (= 2.10) involving deuterated anilines $[XC_6H_4NH_2(D_2)]$ for the anilinolysis, and exceptionally fast rate and biphasic concave downward free energy correlation for the pyridinolysis.

• Bulletin of the Korean Chemical Society
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• v.32 no.4
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• pp.1138-1142
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• 2011

Kinetic studies for the reactions of Y-aryl phenyl chlorothiophosphates with X-pyridines have been carried out in acetonitrile at $35.0^{\circ}C$. The Hammett and Bronsted plots for substituent X variations in the nucleophiles are biphasic concave upwards with a break point at X = 3-Ph, while the Hammett plots for substituent Y variations in the substrates are biphasic concave downwards (and partially upwards) with a break point at Y = H. The signs and magnitudes of the cross-interaction constant (${\rho}_{XY}$) are strongly dependent upon the nature of substituents, X and Y. The proposed mechanism is a stepwise process with a rate-limiting step change from bond breaking with the weaker electrophiles to bond formation with the stronger eletrophiles. The nonlinear free energy correlations of biphasic concave upward plots for substituent X variations in the nucleophiles are rationalized by a change in the attacking direction of the nucleophile from a backside with less basic pyridines to a frontside attack with more basic pyridines.

• Journal of Periodontal and Implant Science
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• v.35 no.4
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• pp.1097-1108
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• 2005

The present study was to determine the influence of micro-macro biphasic calcium phosphate(MBCP) on proliferation and differentiation of human marrow-derived mesenchymal stem cells. Primary stem cells were cultured from bone marrow and 3-4 passaged cells were used. This study tested the proliferative effects by cell counting. Collagen sythensis, alkaline phosphatase activity, expression of osteocalcin and bone sialoprotein by Western blot analysis were evaluated. The cellular proliferation of ASC was not influenced by MBCP. Collagen synthesis of ASC cultured on MBCP significantly increased at 5th and 7th days(p<0.05). The ALP activity in ASC cultured on MBCP significantly increased at 5th and 7th days(p<0.05). The expression of OC and BSP incresaed in ASC cultured on MBCP. These results suggest that MBCP may stimulates the osteoblastic activity of ASC.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.42
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• pp.34.1-34.7
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• 2020

Purpose: The purpose of this study was to evaluate the influence of biodegradable polycaprolactone membrane on new bone formation and the biodegradation of biphasic alloplastic bone substitutes using animal models. Materials and methods: In this study, bony defect was formed at the canine mandible of 8 mm in diameter, and the defects were filled with Osteon II. The experimental groups were covered with Osteoguide as barrier membrane, and the control groups were closed without membrane coverage. The proportion of new bone and residual bone graft material was measured histologically and histomorphometrically at postoperative 4 and 8 weeks. Results: At 4 weeks, the new bone proportion was similar between the groups. The proportion of remaining graft volume was 27.58 ± 6.26 and 20.01 ± 4.68% on control and experimental groups, respectively (P < 0.05). There was no significant difference between the two groups in new bone formation and the amount of residual bone graft material at 8 weeks. Conclusion: The biopolymer membrane contributes to early biodegradation of biphasic bone substitutes in the jaw defect but it does not affect the bone formation capacity of the bone graft.

• Journal of the Korean Crystal Growth and Crystal Technology
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• v.21 no.4
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• pp.169-174
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• 2011

Nanoscale-biodegradable behaviors of synthesized biphasic calcium phosphates (BCP) powders were characterized using X-ray diffraction (XRD), lattice parameter, and field emission microscopy (FE-SEM). The calcined BCP powders in vitro tested in Hank's balanced salt solution (pH = 7.4, $36.5^{\circ}C$) for 3 weeks. The calculated unit cell parameters for BCP have shown lattice distortion and expansion as irregular changes in the a and c-axis after in vitro.