• Applied Microscopy
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• v.40 no.2
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• pp.73-80
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• 2010

Liver regeneration is a result of highly coordinated proliferation of hepatocytes and nonparenchymal liver cells. Partial hepatectomy (PH) is the most often used stimulus to study liver regeneration because, compared with other methods that use hepatic toxins, it is not associated with the tissue injury and inflammation, and the initiation of the regenerative stimulus is precisely defined. Granulocyte macrophage-colony stimulating factor (GM-CSF), which is a cytokine able to regulate the proliferation and differentiation of epithelial cells, was first identified as the most potent mitogen for bone marrow. Particularly, rrhGM-CSF, which is highly glycosylated and sustained longer than any other types of GM-CSF in the blood circulation, was specifically produced from rice cell culture. In this experiment, effects of rrhGM-CSF administration were evaluated in the regenerating liver after 78% PH of rats. Morphological changes induced by PH were characterized by destroyed hepatocyte plate around the central vein and enlarged nuclear cytoplasmic ratio and increased hepatocytes with two nuclei. And then, proliferation of liver cells (parenchymal and nonparenchymal) and rearrangement of plates and lobules seemed to be carried out during liver regeneration. These alterations in the experimental group preceded those of the control. Since proliferating cell nuclear antigen (PCNA) is known to be a nuclear protein maximally elevated in the S phase of proliferating cells, the protein was used as a marker of liver regeneration after PH in rats. PCNA levels by western blot analysis and immunohistology were compared between the two groups. PCNA protein expression of two groups at 12 hr and 24 hr after injury showed similar pattern. The protein expression showed the peak at 3 days in both groups, however, the protein level of the experimental group was higher than that of the control. On immunohistochemical observations, the reaction product of PCNA was localized at the nuclei of proliferating cells and the positive reaction in experimental group at 3 days was clearly stronger than that in control group. The results by Western blotting and immunohistology for PCNA showed similar pattern in terms of the protein levels. In conclusion, rrhGM-CSF administration during liver regeneration after 78% PH accelerated breakdown and restoration of the hepatic plate and expression of PCNA. These results suggest that rrhGM-CSF might play an important role during liver regeneration in rats.

• Journal of the korean academy of Pediatric Dentistry
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• v.26 no.4
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• pp.652-663
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• 1999

Craniosynostosis, the premature fusion of cranial sutures, presumably involves disturbance of the interactions between different tissues within the cranial sutures. Interestingly, point mutaions in the genes encoding for the fibroblast growth factor receptors(FGFRs), especially FGFR2, cause various types of human craniosynostosis syndromes. To elucidate the function of these genes in the early morphogenesis of mouse cranial sutures, we first analyzed by in situ hybridization the expression of FGFR2(BEK) and osteopontin, an early marker of osteogenic differentiation, in the sagittal suture of calvaria during embryonic(E15-E18) and postnatal stage(P1-P3). FGFR2(BEK) was intensely expressed in the osteogenic fronts, whose cells undergo differentiation into osteoprogenitor cells that ultimately lay down the bone matrix. Osteopontin was expressed throughout the parietal bones excluding the osteogenic fronts, the periphery of the parietal bones. To further examine the role of FGF-mediated FGFR signaling in cranial suture, we did in vitro experiments in E15.5 mouse calvarial explants. Interestingly, implantation of FGF2 soaked beads onto both the osteogenic fronts and mid-mesenchyme of sagittal suture after 36 hours organ culture resulted in the increase of the tissue thickness and cell number around FGF2 beads, moreover FGF4-soaked beads implanted onto the osteogenic fronts stimulated suture closure due to an accelerated bone growth, compared to FGF4 beads placed onto mid-mesenchyme of sagittal suture and BSA control beads. In addition FGF2 induced the ectopic expression of osteopontin and Msx1 genes. Taken together, these data indicate that FGF-mediated FGFR signaling has a important role in regulating the cranial bone growth and maintenance of cranial suture, and suggest that FGF-mediated FGFR signaling is involved in regulating the balance between the cell proliferation and differentiation through inducing the expression of osteopontin and Msx1 genes.

• Korean Journal of Food Science and Technology
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• v.31 no.1
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• pp.238-245
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• 1999

Antioxidative effects of the water or ethanol extracts from Rhus verniciflua Stokes (RVS) were measured by protection against hydroxyl radicals in mouse brain tissue culture. In the water extracts from RVS, cell viabilities were estimated 60.0, 66.0, 72.0, 84.0 and 90.0% at addition of 1, 2, 4, 7 and $10{\mu}L$, respectively, compared with GO (20 mU/mL) alone. The cell viability in the ethanol extracts was similarly with water extracts. In the antitumor effects, the results showed that percentages of the HeLa cell death were approximately 24% for 12 hrs, 57% for 48 hrs at addition of 10%/well ethanol extracts respectively. To know inhibition of tumor growth, in vivo, mice (BALB/c) were inoculated with 0.25 mL CT-26 $(1{\times}10^6\;cells/mL)$ subcutaneously. After the generation of tumor, the results of RVS extracts (ethanol, water) injection showed generally that the tumor size in BALB/c was reduced. For physicochemical characterization of the RVS extracts, purified substances of water or ethanol extracts were analized with SDS-PAGE and ICP spectrometer. In electrophoresis, gel showed 2 bands (210, 230 KDa). The results of ICP verified that RVS extracts contain $Cu^{2+}$ in both samples. Conclusively, this substance might be a laccase which has a biological effective function, as a natural bioactive substance.

• Applied Biological Chemistry
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• v.39 no.6
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• pp.494-500
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• 1996

Uptake of hen metal ions by water dropwort (Oenanthe stolonifera DC.) and its cadmium-binding protein were studied to probe for good method to remove heavy metal contaminants from environments. The plant was cultured in the culture medium (pH 7.0) containing the various concentrations of $Cd^{2+}$, $Cr^{3+}$ or $Pb^{2+}$, for 3 and 7 days. The residual heavy metals deposited in roots linearly increased as the metal ions concentration increased up to 17 ppm for $Cd^{2+}$, 20 ppm for $Cr^{3+}$ and 50 ppm for $Pb^{2+}$. Above these concentrations, the plant growth was inhibited and the uptake rates of the metal ions decreased. The heavy metals absorbed by the plant were mostly deposited in roots. In particular, the residual concentration of lead in roots was about four times higher than those of cadmium and chromium. When cultured in the medium containing 20 ppm of each metal ion, 80% of cadmium, 90% of cromium and 96% of lead were deposited in roots out of the total residual metal ions in the plant. These values correspond to 6.1 mg of cadmium, 5.2 mg of chromium and 23.6 mg of lead per one gram of roots tissue on a dry weight basis. A cadmium-binding protein was partially purified by extraction, gel filtration and DEAE-Cellulose chromatography from water dropworts that was grown in the medium containing 20 ppm $Cd^{2+}$. The purified protein was a single band on SDS- and non-denaturing- polyacrylamide gel electrophoresis. Its molecular mass was estimated to be ca. 5,000 dalton by gel filteration. Analysis of amino acid composition of the protein indicated that it had a typical amino acid composition of heavy metal-binding protein in that it contained 27% of acidic amino acids and 9.9% of cysteine. However, it is likely that the protein is a new plant metal-binding protein, since its amino acid composition is somewhat different from those of phytochelatins that have been known so far.

• Korean Journal of Hospice Care
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• v.6 no.1
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• pp.1-11
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• 2006

There are various understandings how to define death. In the context of medicine, death is defined as the irreversible change of the tissue according to the cessation of circulation and respiration. According to the psychologists, a person need to accept the finiteness as a human being and remain conscious that the death is not avoidable. And they say if a person doesn't regard death as unavoidable reality of life he or she will not confront the humanistic death and after all will die like animals. In philosophy, death is viewed as an unwelcome reality in the end of the journey of life. Sociologists usually understand that the society is the organization composed with living persons and human beings which construct and transmit the culture from generation to generation between the both ends of life and death. In society, the generation is changed, maintained, and developed through the phenomenon of death. Although death of human being is natural event in society, the death of a specific person brings a sense of loss, crisis, and anxiety to the communities like family, regional society, nation, and the world. In this context, death is not confined to personal dimension and it can be regarded as a social problem. It is valuable to summarize the religious perspectives on the meaning of death for the better hospice care. In shamanism, there are basic idea that although the flesh of human being disappears, soul never die. If human dies, the flesh of human being disappears but soul never disappear and come back to the origin of soul as it is called chaos. So in shamanism, it is said that shaman can solve the mortified feeling, restore the broken harmony, send the soul to comfortable space- the origin, and guarantee the blessing of descendents. Buddhists regard the death as an essential component through the cycles of life. Through this cycle, human being exits as an endlessly transmigrating being and the death is just a restoration to the original status. In Confucianism, the view on the death based on the philosophy of the "Yin and Yang" and "Five elements". In Buddhist tradition, many believers said the philosophy of "Death is the same as life". Unlike usual thoughts that a god governs "life and death" and "fortune and misfortune", Confucianists deny the governance of a god and emphasize the natural orders in which every phenomenon in the world moves according to the principle. Confucianists understand the death as a natural order with this principle. In Confucianists' belief, the essence of human being remains in their own descendent's lives after the death of ancestor, so in Confucianism there is no concept of immortality of the soul. In the history of Christianity, death has been defined generally as the separation of the immortal soul from the mortal body. In the earlier days of Old Testament, the death is regarded as a disappearance of just a flesh and human never disappear and always live in the relationship with God. Later days in Old Testament, we can find the growing concern for the life after the death because of the entrance of the theodicy. In the New Testament, the death is not regarded as the normal process of the human life and regarded as the abnormal status in which death come to human because of sin as a decisive factor and it should be conquered. In fact, the most of us afraid death because not of the fear of death itself but of the sense of the emptiness and regrets. so many people often make the monument hoping to live forever. But Christian usually regard this behavior as a sinful act because human being usually think themselves as a master of their life and attempt to become immortal in this kind of trial mortal. But if we live with God, we cannot confront such a condition because we aware limits as a mortal human being and entrust everything on Him and want to live according to His guidance. Therefore, in the Christian tradition, the death is regarded as accomplishment of life, fruits of life, invitation to the eternal life, and the last stage of human growth. For human being, the death is the great step of maturation as a human in the final stage of life.

• Tuberculosis and Respiratory Diseases
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• v.58 no.4
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• pp.375-384
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• 2005

Background and Aims : Pre-induction of heat shock protein 70 (HSP70) is known to effectively attenuate the lipopolysaccharide (LPS)-induced inflammatory response in lung tissue. However, it is unclear if HSP70 induction after LPS exposure attenuates the subsequent LPS-induced inflammatory response in alveolar epithelial cells. This study examined the effects of HSP70 induction after LPS exposure on the IL-6 production and the cell viability after a subsequent LPS challenge in murine alveolar epithelial cells, and investigated whether or not HSP70 itself may be involved in those effects. Methods : Murine alveolar epithelial cells were cultured and divided into two groups; the Non-Pre-LPS group without a LPS pre-treatment and the Pre-LPS group with a LPS pre-treatment. Each group was subdivided into the following four subgroups: subgroup C (control), subgroup Q (quercetin), subgroup HSP70 (HSP70 induction), and subgroup HSP70-Inh (HSP70 inhibition). HSP70 expression, which was induced by sodium arsenite and inhibited by quercetin, was analyzed by western blot analysis. The IL-6 levels in the culture supernatant were measured by ELISA, and the cell viability was measured using a simplified MTT assay. Results : The IL-6 levels were lower in subgroup HSP70 than in subgroup C (P<0.01), and were higher in subgroup HSP70-Inh than in subgroup HSP70 in both the Non-Pre-LPS and Pre-LPS groups (P<0.05, P<0.01). The cell viability tended to decrease in the Pre-LPS group compared with the Non-Pre-LPS group. While the cell viability was higher in subgroups Q, HSP70, and HSP70-Inh than in subgroup C in the Non-Pre-LPS group (P<0.05, P<0.05, P<0.01), there was no difference in cell viability among the subgroups in the Pre-LPS group. Conclusion : HSP70 induction after a LPS pre-treatment in murine alveolar epithelial cells inhibits the subsequent LPS-induced IL-6 production without affecting the cell viability, and HSP70 by itself may play an important role in this proccess.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.26 no.1
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• pp.110-114
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• 1981

These experiments were carried out to define the effects of NAA, 2, 4-D and Benzyladenine on the callus induction and the organ differentiation from the explants and to find out the vegetative propagation method of Korean ginseng. The results obtained are summarized as follows; 1. NAA was significantly effective in forming roots from the ginseng stem segment and the number of roots was increased by increasing NAA concentration in the medium. The roots were formed from both distal and proximal ends of the ginseng stem segments grown on the medium containing more than 2mg/L of NAA. 2. The amount of callus growth increased proportionatly with NAA concentration in the range of 4.0mg per liter in the medium. The callus was easly induced from stem segment than leaf segment and 2, 4-D was more effective in callus induction and growth than NAA. 3. The benzyladenine showed the significant inhibition effect in forming roots from ginseng explant. The callus was not induced with BA alone, but in BA and 2, 4-D or BA and NAA added medium, the callus was easily induced and its growth was also accelerated. The interaction effects between 2, 4-D and BA on the callus induction and growth were significantly higher than those between NAA and BA. 4. As the ginseng embryos were cultured on the M.S. medium supplemented with 2mg per liter NAA, number of shoots was significantly increased and the percentage of embryo which had shown more than 4 shoots later was 22.2%. On the medium containing 8mg per liter NAA, the ginseng embryo showed the normal growth of shoots and leaves, but increased roots and callus induction on the basal part of shoots. 5. When the shoots with 3 leaflets were cut in 1.5cm long and grown on the Blayde's medium containing NAA 1.0mg per liter, roots were formed at the proximal end of shoot, and a new ginseng seedling was successfully obtained.

• Korean Journal of Medicinal Crop Science
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• v.4 no.1
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• pp.78-85
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• 1996

The effects of growth regulators, sucrose and gelling agents were investigated to increase the efficiency of the callus growth and plant regenerarion in tissue culture of Angelica koreana Max. The fresh weight and dry weight of subcultured callus was highest in MS medium supplemented with 1 mg/l 2,4-D. Callus growth was excellent in 2% sucrose, but it was inhibited in propotion to sucrose content. Effect of gelling agents on callus growth was highest on 1.2% agar and 0.4% Gelrite medium, respectively. The browning of callus was protected on the media supplemented with 10 mg/l ABA and 5 or 10 mg/l $AgNO_3$. In the callus induction and growth from the peduncle of immature inflorescence, 2,4-D was more effective than NAA, and the frequency of callus induction was highest as 81.7% in 2 mg/l 2,4-D. Plant was not regenerated from the callus derived from young leaf. Somatic embryos were developed from the surface of callus drived from the peduncle of immature inflorescence in the medium containing 0.5 mg/l 2,4-D, 1 mg/l kinetin, 5 mg/l ABA and 5 mg/l $AgNO_3$. Plants were developed from the matured somatic embryos in the medium supplemented with 0.2 mg/l 2,4-D and 1 mg/l kinetin.

• Korean Journal of Veterinary Research
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• v.25 no.1
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• pp.7-17
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• 1985

Many investigators have been pursuing various attempts so far to produce hepatitis B surface antigen(HBsAg) vaccines using the techniques such as isolation from plasma of chronic HBsAg carrier, recombinant DNA technique or preparation of synthetic peptides specific for immunogenic determinants. Hepatitis B virus can not grow on any cell lines by the tissue culture technique at the present time. The plasma of chronic HBsAg carrier is expensive and its source is limited. The HBsAg from the recombinant DNA technique gave still very low yield. Another approach, therefore, has been initiated to develop a synthetic hepatitis B virus vaccine. The possible use of several distinct synthetic vaccines in prophylaxis can be facilitated by availability of full synthetic immunogens. Peptides synthesized for potential application as antiviral vaccines have been mostly tested in the form of conjugates with carrier proteins, although the free synthetic peptide can be immunogenic. To understand basic knowledges on the antigenicity and immunogenicity of a synthetic peptide specific for major immunogenic determinant of HBsAg, a nonapeptide, $H_2N^{139}Cys-Thr-Lys-Pro-Thr-Asp-Gly-^{146}Asn-Aba$ COOH, which corresponds to HBsAg amino acid residues 139 to 147, was synthesized by the Merrifield's solid-phase method with a slight modification. The antigenicity and immunogenicity of this specific synthetic peptide were examined comparing with purified plasma-derived natural HBsAg. The results obtained are as follows; 1. The peptide synthesized showed the identical amino acid composition to the theoretical value. The degree of purification and molecular weight were acertained by methods of high performance liquid chromatography and mass spectrometry. 2. Using m-maleimidobenzoyl-N-hydroxysuccinimide ester as a conjugating agent, the synthetic peptide was conjugated to rabbit albumin and ${\gamma}$-globulin, tetanus and diphtheria toxoids, and keyhole limpet hemocyanin. Their conjugation yields were 8.3, 9.5, 15.8, 13.5, and 11.2%, respectively. 3. The natural HBsAg was purified from plasma of chronic HBsAg carrier. By the electron microscopic observation of the purified natural HBsAg preparation, no Dane particles were observed and the preparation showed negative DNA polymerase activity. 4. Antigenicity of the synthetic peptide and the plasma-derived natural HBsAg was determined by competition radioimmunoassay using $^{125}I$-natural HBsAg. Their 50% inhibitions appeared as $90{\mu}g/ml$ and $0.12{\mu}g/ml$ for the synthetic peptide and the natural HBsAg, respectively. This indicates that the former was about 750-fold less antigenic than the latter. 5. Immunogenicity of the synthetic peptide was determined by administering the peptide-carrier conjugates into rabbits with and without Freund's complete adjuvant. Regardless the carrier proteins and adjuvant, positive immune responses to the synthetic peptide were observed. The higher antibody titers, however, were shown in the groups administered with Freund's complete adjuvant. 6. Immunizing dose 50% in mice of the various peptide-carrier conjugates was 5.47, 6.00, 65.16, 31.25 and $13.03{\mu}g/dose$ for rabbit albumin and ${\gamma}$-globulin, tetanus and diphtheria toxoids, and keyhole limpet hemocyanin, respectively, while the natural HBsAg showed $0.65{\mu}g/dose$. 7. It was postulated that homologous proteins prefer to heterologous ones as the carriers.

• Korean Journal of Weed Science
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• v.8 no.1
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• pp.53-58
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• 1988

This study was carried out to determine effect of butachlor [N.-(buthoxymethyl)-2-chloro-N-(2,6-diethylphenyl) acetamide] and bialaphos [2-amino-4(hydroxy)(methyl) phosphionyl] butyryl-alanylalanine sodium salt on the germination of tobacco seed, induction and growth of callus from tobacco. Further, fatty acids and ammonia content of tobacco calli were determined. Bialaphos had no effect on tobacco seed germination, but the growth of seedling was markedly affected by an application of 10 ppm bialaphos. However, regardless of varieties tested, tobacco seed germination was completely inhibited by $5{\times}10^{-5}M$ of butachlor. At an application of $5{\times}10^{-5}M$ butachlor, tobacco seeds were to some extent germinated and showed further growth. Hyangcho among varieties tested, showed the most tolerant response to butachlor. In induction of callus from various tobacco varieties and their growth, aromatic type of tobacco varieties exhibited the most tolerance against bialaphos. However, no distinct varietal differences were determined in the treatment of butachlor. The major fatty acids identified in tobacco calli were palmitic, oleic and linoleic acid. No marked difference in terms of fatty acids was observed among tobacco varieties used, but it was observed that there was the higher ratio of quantity in unsaturated fatty acids over saturated one, bialaphos treatment accumulated about 9 times higher ammonia content than that of the untreated control, giving an evidence that bialaphos might inhibit glutamine synthetase activity.