• Asian Pacific Journal of Cancer Prevention
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• 제13권8호
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• pp.4031-4036
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• 2012

Background: The negative signaling provided by interactions of the co-inhibitory molecule, programmed death-1 (PD-1), and its ligands, B7-H1 (PD-L1) and B7-DC (PD-L2), is a critical mechanism contributing to tumor evasion; blockade of this pathway has been proven to enhance cytotoxic activity and mediate antitumor therapy. Here we evaluated the anti-tumor efficacy of AAV-mediated delivery of the extracellular domain of murine PD-1 (sPD-1) to a tumor site. Material and Methods: An rAAV vector was constructed in which the expression of sPD-1, a known negative regulator of TCR signals, is driven by human cytomegalovirus immediate early promoter (CMV-P), using a triple plasmid transfection system. Tumor-bearing mice were then treated with the AAV/sPD1 construct and expression of sPD-1 in tumor tissues was determined by semi quantitative RT-PCR, and tumor weights and cytotoxic activity of splenocytes were measured. Results: Analysis of tumor homogenates revealed sPD-1 mRNA to be significantly overexpressed in rAAV/sPD-1 treated mice as compared with control levels. Its use for local gene therapy at the inoculation site of H22 hepatoma cells could inhibit tumor growth, also enhancing lysis of tumor cells by lymphocytes stimulated specifically with an antigen. In addition, PD-1 was also found expressed on the surfaces of activated CD8+ T cells. Conclusion: This study confirmed that expression of the soluble extracellular domain of PD-1 molecule could reduce tumor microenvironment inhibitory effects on T cells and enhance cytotoxicity. This suggests that it might be a potential target for development of therapies to augment T-cell responses in patients with malignancies.

• Korean Parent-Child Health Journal
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• 제1권
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• pp.75-87
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• 1998

These days social and economical changes have influence on the structure of family and the role of family members. Working mothers and widowers with children are increasing because of economical difficulties. Support from relatives are decreasing because of the conspicuous trend toward nuclear families. According to these reasons androgynous fathers are required. Today's fathers in Korea socially and culturally have learned about traditional parenting, but they are changing their fathering styles to meet the demands of the times. However they don't have their own fathering models. Therefore nurses who hold an advantageous position to teach and support from clinic have to encourage them to care their infants. The purposes of this study were to define father's care giving in infancy, understand influencing factors on fathering, and the differences between fathering and mothering, then contribute to nursing implementation for supporting fathers. This study was designed to review references about father's care giving. The results were follows: Six aspects of parent participation were direct care. indirect care, play, decision-making concerning the child, amount of time of sole responsibility for the child and overall availability to the child. Direct care involved feeding, bathing, going to child if child awakens. dressing, putting child to bed, taking child to doctor, nurse, or dentist, transporting child to and from sitter, day care, or school, washing child's hair. Indirect care involved cleaning up after child, preparing child's food, fixing child's broken playthings, washing child's clothes, arranging baby-sitting, shopping for child's toys and clothes, transporting baby-sitter to and from your home. Young fathers were gradually participating in direct care like feeding, taking child to doctor. Father's care giving stimulated mothering and promoted parent-infant relationship. Influencing factors of fathering would be divided into father characteristics, surrounding factors, infant attributes. Father characteristics were age, role perception, relationship with parent. Surrounding factors were the opportunity of early contact, support system, spouse's expectation, marital adjustment, feeding type, past experience of care giving. Infant attributes were temperament, behavior, age, sex. The differences between fathering and mothering were reviewed. Fathers were poor at care giving. but their caring was similar to mother's. This subtle difference positively worked upon infant's growth and development. On the basis of these theoretical data, nurses can empower fathers to cooperate with mothers in caring infants.

• Biomolecules & Therapeutics
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• 제19권2호
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• pp.187-194
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• 2011

Atherosclerosis and post-angiography restenosis are associated with intimal thickening and concomitant vascular smooth muscle cell (VSMC) proliferation. Obovatol, a major biphenolic component isolated from the Magnolia obovata leaf, is known to have anti-inflammatory and anti-tumor activities. The goal of the present study was to enhance the inhibitory effects of obovatol to improve its potential as a preventive or therapeutic agent in atherosclerosis and restenosis. Platelet-derived growth factor (PDGF)-BB-induced proliferation of rat aortic smooth muscle cells (RASMCs) was examined in the presence or absence of a newly synthesized obovatol derivative, OD78. The observed anti-proliferative effect of OD78 was further investigated by cell counting and [$^3H$]-thymidine incorporation assays. Treatment with 1-4 ${\mu}M$ OD78 dose-dependently inhibited the proliferation and DNA synthesis of 25 ng/ml PDGF-BB-stimulated RASMCs. Accordingly, OD78 blocked PDGF-BB-induced progression from the $G_0/G_1$ to S phase of the cell cycle in synchronized cells. OD78 decreased the expression levels of CDK4, cyclin E, and cyclin D1 proteins, as well as the phosphorylation of retinoblastoma protein and proliferating cell nuclear antigen; however, it did not change the CDK2 expression level. In addition, OD78 inhibited downregulation of the cyclin-dependent kinase inhibitor (CKI) $p27^{kip1}$. However, OD78 did not affect the CKI $p21^{cip1}$ or phosphorylation of early PDGF signaling pathway. These results suggest that OD78 may inhibit PDGF-BB-induced RASMC proliferation by perturbing cell cycle progression, potentially through $p27^{kip1}$ pathway activation. Consequently, OD78 may be developed as a potential anti-proliferative agent for the treatment of atherosclerosis and angioplasty restenosis.

• The Journal of Advanced Prosthodontics
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• 제3권3호
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• pp.152-160
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• 2011

PURPOSE. This study was to investigate the effects of recombinant human platelet-derived growth factor (rhPDGF-BB) and heparin to titanium surfaces for enhancement of osteoblastic functions and inhibition of inflammation activity. MATERIALS AND METHODS. The anodized titanium discs, not coated with any material, were used as a control group. In heparinized-Ti group, dopamine was anchored to the surface of Ti substrates, and coated with heparin. In PDGF-Ti group, rhPDGF-BB was immobilized onto heparinized Ti surface. The surface morphologies were investigated by the scanning electron microscope in each group. The release kinetics of rhPDGF-BB were analyzed, and cytotoxicity tests for each group were conducted. The biocompatibilities were characterized by measuring cell proliferation, alkaline phosphatase activity, and calcium deposition using MG-63 cells. Statistical comparisons were carried out by one-way ANOVA tests. Differences were considered statistically significant at $^*$P<.05 and $^{**}$P<.001. RESULTS. The combination of rhPDGF-BB and heparin stimulated alkaline phosphatase activity and OCN mRNA expression in osteoblastic cells ($^*$P<.05 and $^{**}$P<.001). MG-63 cells grown on PDGF-Ti had significantly higher amounts of calcium deposition than those grown on anodized Ti ($^{**}$ P<.001). Heparinized Ti was more anti-inflammatory compared to anodized Ti, when exposed to lipopolysaccharide using the transcript levels of TNF-${\alpha}$ and IL-6 of proinflammatory cytokine ($^*$P<.05 and $^{**}$P<.001). CONCLUSION. The result of this study demonstrated that the incorporation of rhPDGF-BB and heparin onto Ti surface enhanced osteoblastic functions and inhibited inflammation.

• Asian-Australasian Journal of Animal Sciences
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• 제9권5호
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• pp.583-590
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• 1996

The purpose of this study was to evaluate some factors in the bovine embryonic development from one-cell to blastocyst using modified synthetic oviduct fluid medium (mSOFM), after maturation and in vitro fertilization of the oocytes. The embryonic development to the blastocyst stage was assessed at 7-10 days after in vitro fertilization, and the total cells in the blastocysts were counted by staining nuclei with fluorochrome. Some commercial calf sera (CS) and a superovulated cow serum had different effects on the embryonic development to the blastocyst stage (8.6-21.4%), dependent upon their product lots, although the development might not be affected at least by serum progesterone levels. ${\beta}$-Mercaptoethanol (${\beta}$-ME) supplemented into mSOFM was effective to the embryonic development (27.8%), as well as the co-culture system with cumulus cells (19.5%). In a serum- and feeder cell-free culture using mSOFM containing several growth factors and ${\beta}$-ME instead of CS plus co-cultured cumulus cells, bovine serum albumin (BSA, fraction V), but not polyvinyl alcohol (PVA), was highly effective in embryonic development to the blastocyst stage, almost comparable to CS in the serum-contained culture (CS, BSA and PVA; 27.8, 19.5 and 5.7%, respectively). However, fatty acid free BSA rather reduced the number of developed blastocysts, compared with fraction V BSA (7.3 vs 29.4%). In the serum- and feeder cell-free culture, supplement of glucose to the medium (final 2.0 mM) stimulated the cell proliferation of developing embryos 120 hr after in vitro fertilization. These results indicated that a serum-free medium supplemented with ${\beta}$-ME could successfully support the development of bovine one-cell embryos to the blastocyst stage. Moreover, supplement of glucose and fatty acids to the medium might support preferably the development and cell proliferation of embryos.

• BMB Reports
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• 제35권3호
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• pp.330-336
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• 2002

The lgtB genes that encode $\beta$-1,4-galactosyltransferases from Neisseria meningitidis ATCC 13102 and gonorrhoeae ATCC 31151 were isolated by a polymerase chain reaction using the pfu DNA polymerase. They were expressed under the control of lac and T7 promoters in Escherichia coli M15 and BL21 (DE3). Although the genes were efficiently expressed in E. coli M15 at $37^{\circ}C$ (33 kDa), most of the $\beta$-1,4-galactosyltransferases that were produced were insoluble and proteolysed into enzymatically inactive polypeptides that lacked C-terminal residues (29.5 kDa and 28 kDa) during the purification steps. When the temperature of the cell growth was lowered to $25^{\circ}C$, however, the solubility of the $\beta$-1,4-galactosyltransferases increased substantially. A stable N-terminal his-tagged recombinant enzyme preparation could be achieved with E. coli BL21 (DE3) that expressed lgtB. Therefore, the cloned $\beta$-1,4-galactosyltransferases were expressed under the control of the T7 promoter in E. coli BL21 (DE3), mostly to the soluble form at $25^{\circ}C$. The proteins were easily purified to homogeneity by column chromatography using Ni-NTA resin, and were found to be active. The galactosyltransferases exhibited pH optimum at 6.5-7.0, and had an essential requirement for the $Mn^{+2}$ ions for its action. The $Mg^{+2}$ and $Ca{+2}$ ions showed about half of the galactosyltransferase activities with the $Mn^{+2}$ ion. In the presence of the $Fe^{+2}$ ion, partial activation was observed with the $\beta$-1,4-galactosyltransferase from N. meningitidis(64% of the enzyme activity with the $Mn^{+2}$$Ni^{+2}$, $Zn^{+2}$, and $Cu^{+2}$ ions could not activate the $\beta$-1,4-galactosyltransferase activity. The inhibited enzyme activity with the $Ni^{+2}$ ion was partially recovered with the $Mn^{+2}$$Fe^{+2}$, $Zn^{+2}$, and $Cu^{+2}$ ions, the $Mn^{+2}$$\beta$-1,4-galactosyltransferase activity was 1.5-fold stimulated with the non-ionic detergent Triton X-100 (0.1-5%).

• Journal of Korean Society of Forest Science
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• 제89권1호
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• pp.116-125
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• 2000

This study was conducted to investigate the effects of CEPA(2-chloroethyl phosphonic acid) on bark physiology and anatomy of lactree(Rhus verniciflua Stokes). Sample trees of similar size and growth rate were selected from 7-year-old lactree plantation located in Hyengsung-kun, Kangwon-do. Lanolin pastes containing 0.1, 1, or 10% CEPA were put into the bark-removed hole made by corer(${\phi}1cm$) on the main stem at 1.2m above the ground on June 16, 1995. Five weeks after application of CEPA, bark thickness was markedly increased as a result of the increase in the amount of phloem and intercellular spaces, and correlated with the increased production of urushiol. By the application of 10% CEPA, bark thickness was increased approximately 2.5 times, and the urushiol content within bark was increased 2.8 times compared to that of untreated trees because CEPA stimulated the accumulation of urushiol within bark. Treatment of 10% CEPA also increased the size and the total number of secretory canals, and induced an increase in ray width. The phloem parenchyma cells of CEPA-treated trees were well-developed and closely packed with little intercellular space.

• Journal of the Korean Society of Food Science and Nutrition
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• 제34권10호
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• pp.1579-1585
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• 2005

The aim of this study was to propose utilization of tofu whey concentrates separated by ultrafiltration (UF) for yogurt production. The curd yogurt was prepared from whole milk added with skim milk Powder in which UF powder was substituted 0$\%$, 6.25$\%$, 12.5$\%$ or 25$\%$ for skim milk powder. The quality characteristics of yogurt in terms of pH, titratable acidity, viscosity, color and viable cell counts were evaluated. There were no significant differences in pH and titratable acidity between control (yogurt added with only skim milk powder) and yogurt added with UF powder after 24 hr of fermentation at 37$^{\circ}C$. Apparent viscosity of yogurt added with UF powder was lower (2,623 $\∼$ 3,189 cps) than that of the control yogurt (3,196 cps). Lightness and redness value of yogurt added with UF powder were not significantly different from control yogurt, while yellowness value increased as the amount of UF powder increased. Addition of UF powder stimulated the growth of lactic acid bacteria. Gas chromatographic analysis detected acetaldehyde, diacetyl, and organic acids, and more volatile compounds were detected in yogurt added with UF powder. Sensory analysis showed that yogurt added with UF powder were evaluated as acceptable as control yogurt. Therefore, UF powder could be substituted for skim milk powder without depressing yogurt qualify.

• Korean Journal of Environmental Agriculture
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• 제20권2호
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• pp.116-121
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• 2001

To observe the stimulating effects of low dose ${\gamma}$ - radiation on the dormancy breaking and physiological activity, potato (Solanum tuberosum L. cv. Dejima) were irradiated at the dose of $0.5{\sim}30$ Gy. Low dose ${\gamma}$ - radiation exhibited promoting effects on the sprouting rate within the range of optimum dose of 1 Gy and 2 Gy and on the sprout length within the range of optimum dose of 2 Gy and 4 Gy. Regardless of storage duration and irradiation doses, the number of sprouts was enhanced. The number of sprouts, which were reserved for 15 days after production (DAP), were significantly increased under 4, 8 and 16 Gy irradiation. The growth of 45 DAP sprouts was extremely stimulated under 4 Gy irradiation accompanying the increase of peroxidase activity in the plantlet. In this study, it was also suggested that the activities of antioxidative enzymes of potato plantlets were not related to the irradiation dose during the plantlet development with the exception of decrease in catalase activity.

• Journal of Animal Science and Technology
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• 제48권3호
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• pp.415-424
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• 2006

The purpose of this study was to investigate the fungal growth and enzyme production under different carbohydrate substrate conditions. The anaerobic fungus Neocallimastix sp. NLRI-3 isolated from the rumen of Korean native goat was incubated with different carbohydrate media containing 0.2% of glucose, starch, rice straw, filter paper, carboxymethyl cellulose(CMC), Sigmacell cellulose, xylan or xylose, respectively. The culture head gas production was the highest in the culture of filter paper medium, and the lowest in the culture of CMC medium at 96h incubation (P<0.05). The fungal zoospore production reached peak at 72h incubation, and its number was the highest in rice straw medium among the treatments (P<0.05). At 96h incubation, carboxymethyl cellulase(CMCase) activity was the highest in the culture of filter paper medium and the lowest in the culture of starch medium (P<0.05). While xylanase activity was the highest in the culture of rice straw medium and the lowest in the culture of xylose medium(P<0.05) at 72h incubation. There were no differences in culture supernatant protein expression among the treatments. However, the patterns of enzyme expression were different among the treatments with zymogram analysis. Six CMCases and 4 xylanase were detected from the results of zymogram analysis. Therefore the present study indicating that the fungal enzyme expression could be stimulated with insoluble substrates in the culture medium.