• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2002.11a
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• pp.116-116
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• 2002

Further development of reconstructed embryos may be dependent upon the synchronization of donor nucleus and recipient cytoplasm at cell fusion, To control the synchronization of donor and recipient cells, the enucleated MII arrested oocytes are artificially stimulated prior to embryo reconstruction. Destruction of cyclin B results in the exit of cells from M-phase of cell cycle. This study was designed to investigate the effects of single or combined stimulation affected cyclin B1 mRNA and protein levels in mouse oocytes. The oocyte activation was induced by 7% ethanol or 10$\mu\textrm{g}$/$m\ell$ Ca-ionophore without (single) or with (combined) 10$\mu\textrm{g}$/$m\ell$ cycloheximide. Competitive quantitative PCR for cyclin Bl mRNA and western blot analysis for cyclin B1 protein was preformed in mouse oocytes. Cyclin B1 mRNA level was significantly reduced in single (P<0.05) and combined (P<0.05) stimulation groups. However, this level did not change in non-activated group and increased in intact group. Cyclin B1 protein level was also significantly reduced in both single (P<0.05) and combined (P<0.05) stimulation groups. In conclusion, single and combined stimulation induces the degradation of cyclin B1 mRNA and protein after activation in enucleated mouse oocytes.

• The Plant Pathology Journal
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• v.24 no.3
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• pp.352-356
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• 2008

Root discs of 4-year-old ginseng, Panax ginseng C. A. Meyer, were inoculated with the higher($10^8$ colonyforming units(CFU)/ml) and lower($10^6\;or\;10^5$ CFU/ml) initial inoculum levels of a plant-growth promoting rhizobacterium(PGPR), Paenibacillus polymyxa GBR-1 to examine rot symptom development and bacterial population changes on the root discs. At the higher inoculum level, brown rot symptoms developed and expanded on the whole root discs in which the bacterial population increased continuously up to 4 days after inoculation. In light and electron microscopy, ginseng root cells on the inoculation sites were extensively decayed, which were characterized by dissolved cell walls and destructed cytoplasmic contents. However, no rot symptoms were developed and the bacterial population increased only during the initial two days of inoculation at the lower inoculum level($10^6$ CFU/ml) of P. polymyxa GBR-1. At the lower inoculum level($10^5$ CFU/ml), boundary layers with parallel periclinal cell divisions, structurally similar to wound periderm, were formed internal to the inoculation sites, beneath which the cells were intact containing numerous normal-looking starch granules and no disorganized cell organelles, suggesting that these structural features may be related to the suppression of symptom development, a histological defense mechanism.

• Proceedings of the Korean Society of Applied Pharmacology
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• 2000.04a
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• pp.17-19
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• 2000

Heterotrimeric G Proteins transduce ligand binding to a wide variety of seven transmembrane cell surface receptors into intracellular signals. The currently accepted model for the activation of G protein suggests that ligand-activated receptor accelerates GDP-GTP exchange reactions on the ${\alpha}$ subunit of the heterotrimeric G protein. At least seventeen distinct isoforms of the G${\alpha}$ subunit protein have been identified in mammalian organisms. Among them, the G${\alpha}$q family consists of five members whose ${\alpha}$ subunits show different expression patterns. G${\alpha}$q and G${\alpha}$11 seem to be almost ubiquitously expressed, whereas G${\alpha}$14 is predominantly expressed in spleen, lung, kidney and testis. G${\alpha}$16 and its murine counterpart G${\alpha}$15 are expressed in hematopoietic cells and has been shown to couple a wide variety of receptors to phosphoinositide-specific phospholipase C activity. Beta-isoforms of phospholipase C were shown to be activated by all members of G${\alpha}$q family, i.e., G${\alpha}$q, G${\alpha}$11, G${\alpha}$l4 and G${\alpha}$16 subunits either in reconstitution system. or in experiments using cDNA transfection with intact Cos-7 cells.

• Journal of Veterinary Clinics
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• v.22 no.4
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• pp.439-443
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• 2005

Rejection is one of the life-threatening complications after organ transplantation. An eight-month-old, intact male, mixed breed dog was presented with acute rejection after renal allograft. The heterotopic renal transplantation with bilateral nephrectomy was performed in the dog. The triple drug protocol for immunosuppression was applied for prevention of the acute rejection. Postoperative care was done according to the transplantation protocol of VMTH, Kangwon National University. The dog was euthanized when the serum creatinine concentration exceeded 5 mg/dL followed by tile signs of illness. The transplanted kidney was enlarged. The renal cortex lesions were characterized by necrosis of the renal tubules and the glomeruli. Interstitial lesions were characterized by hemorrhage and severe infiltration of lymphoid cells. Intrarenal arteries showed necrosis of the walls and infiltration of perivascular lymphoid cells. In immunohistochemical (IHC) findings, infiltration of the CD4 and the CD8 positive T lymphocytes was examined. In this case, acute rejection was shown by humoral and cellular immunity on the basis of histopathologic and IHC evaluation.

• BMB Reports
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• v.38 no.4
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• pp.468-473
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• 2005

The energetics of nitrate uptake by intact cells of the halotolerant cyanobacterium Aphanothece halophytica were investigated. Nitrate uptake was inhibited by various protonophores suggesting the coupling of nitrate uptake to the proton motive force. An artificially-generated pH gradient across the membrane (${\Delta}pH$) caused an increase of nitrate uptake. In contrast, the suppression of ${\Delta}pH$ resulted in a decrease of nitrate uptake. The increase of external pH also resulted in an enhancement of nitrate uptake. The generation of the electrical potential across the membrane ($\Delta\psi$) resulted in no elevation of the rate of nitrate uptake. On the other hand, the valinomycin-mediated dissipation of $\Delta\psi$ caused no depression of the rate of nitrate uptake. Thus, it is unlikely that $\Delta\psi$ participated in the energization of the uptake of nitrate. However, $Na^+$-gradient across the membrane was suggested to play a role in nitrate uptake since monensin which collapses $Na^+$-gradient strongly inhibited nitrate uptake. Exogenously added glucose and lactate stimulated nitrate uptake in the starved cells. N, N'-dicyclohexylcarbodiimide, an inhibitor of ATPase, could also inhibit nitrate uptake suggesting that ATP hydrolysis was required for nitrate uptake. All these results indicate that nitrate uptake in A. halophytica is ATP-dependent, driven by ${\Delta}pH$ and $Na^+$-gradient.

• Microbiology and Biotechnology Letters
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• v.23 no.3
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• pp.255-262
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• 1995

Intact cells of Acinetobacter sp. T5-7 completely degraded trichloroethylene (TCE) following growth with phenol. This strain could grow on at least eleven aromatic compounds, e.g., benzaldehyde, benzene, benzoate, benzylalochol, catechol, caffeic acid, 2.4-D, p-hydroxybenzoate, phenol, protocatechuate and salicylate, and did grow on alkane, such as octane. But except phenol, other aromatic compounds did not induced TCE degradation. Phenol biotransformation products, catechol was identified in the culture media. However, catechol-induced cells did not degrade TCE. So we assumed that phenol hydroxylase was responsible for the degradation of TCE. The isolate T5-7 showed growth in MM2 medium containing sodium lactate and catechol rather than phenol, but did not display phenol hydroxyalse activity, suggesting induction of enzyme synthesis by phenol. Phenol hydroxylase activity was independent of added NADH and flavin adenine dinucleotide but was dependent on NADPH addition. Degradation of phenol produced catechols which are then cleaved by meta-fission. We identified catechol-2.3-dioxygenase by active staining of polyacrylamide gel.

• Biomedical Science Letters
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• v.9 no.3
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• pp.111-121
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• 2003

Recombinant single chain Fv (scFv) antibodies offer many advantages over mouse monoclonal antibodies such as faster clearance from blood, improved tumor localization, reduced human anti-mouse antibody (HAMA) response, and the availability to manipulate the scFv through genetic approaches. The recombinant phage display was constructed using lym-l hybridoma cells as a source of genetic starting material. mRNA was isolated from the corresponding antibodies hybridoma cells. VH and VL cDNA were amplified with RT-PCR and linked with ScFv by linker DNA to form ScFv DNA, which then were inserted into phagemid pCANTAB5E. The phage of positive clones selected with tube containing raji lymphoma cell and infected by competent E. coli HB2151 to express soluble scFv. The scFv lym-l was secreted into the cytosol and culture supernatant and shown to be of expected size (approximately 32 kDa) by western blot. An active scFv lym-l could be produced in E. coli with soluble form and high yield from hybridoma cell line, using phage display system. Immunoreactivity indicated that scFv lym1 showed a potential biding affinity against the raji lymphoma cell as its parental antibody (intact lym-l Ab).

• BMB Reports
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• v.43 no.3
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• pp.205-211
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• 2010

Sprouty (Spry) proteins have previously been suggested as negative regulators of the MAPK pathway through interaction with Raf-1. However, the molecular basis of this inhibition has not been elucidated. In this study, we used cells expressing FLAGtagged Raf-1 with point mutations at known phosphorylation sites to reveal that activation of Raf-1 mutants does not correlate with their degree of interaction with Spry2. The association of Raf-1 with Spry2 in intact cells was further corroborated by immunofluorescence colocalization. Additionally, there was no significant change observed in the strength of interaction between Raf-1 mutants and Spry2 after paclitaxel treatment despite differences in the activation levels of these mutants. Thus, our study provides the evidence that Spry2 does not directly regulate Raf-1 kinase activity, but instead acts as a scaffolding protein that assists interactions between Raf-1 kinase and its direct regulators.

• Proceedings of the SCSK Conference
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• 2003.09b
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• pp.200-200
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• 2003

Intact mammalian hair and wool fibres are multi-compartmental composite materials consisting of a sulphur-rich outer protective cuticle layer surrounding elongated, highly keratinized, cortex cells. The cortex cells themselves are made up of crystalline, filamentous, low-sulphur a-helical keratin molecules embedded in a matrix of highly cross-linked, globular high-sulphur keratins. It is the structurally organised and highly disulphide cross-linked nature of these materials that provides them with their remarkable mechanical properties. However these mechanical properties are sensitive to environmental conditions such as water content, temperature and chemical treatment and the importance of their ultra-structural arrangements to overall mechanical properties in different environments is still not fully understood.(omitted)

• Korean Journal of Oriental Medicine
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• v.15 no.3
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• pp.99-105
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• 2009

In this research, the genotoxic effects of Pinelliae Rhizoma (PR) extracts, one of famous herbal agents in Korean medicine were evaluated using the mouse micronucleus test. PR extracts was administered once a day for 2 continuous days by oral gavage to male ICR mice at doses of 2000, 1000, and 500 mg/kg. Cyclophosphamide was used as a known genotoxic agent in a positive control. The appearance of a micronucleus is used as an index for genotoxic potential. No PR extracts treatment-related abnormal clinical signs, body weight changes and mortalities were detected. Significant (p<0.01) increases of the numbers of polychromatic erythrocytes contain micronucleus in prepared bone marrow cells were detected in CPA and PR extracts 2000 mg/kg treated groups as compared with intact control, respectively. The results of intraperitoneal dose mouse bone marrow cell micronucleus test of PR extracts were positive in the present study. It is considered that there were no problems from cytotoxicity of PR extracts tested in this study because the polychromatic erythrocyte ratio was detected as > 0.42 in all tested groups.