• The Korean Journal of Pharmacology
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• v.30 no.2
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• pp.191-203
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• 1994

$K^+$ channel openers (KCOs) are known to have a wide range of effects by opening the $K^+$ channel in plasma membranes of various smooth muscles, cardiac muscle and pancreatic ${\beta}-cell$. In the present study, we investigated the effects of 5 types of KCOs, cromakalim, RP49356, pinacidil, nicorandil and diazoxide on the contractility of isolated rat uterus. All KCOs tested inhibited the uterine contraction induced by 0.2 nM oxytocin in a dose-dependent manner. Individual KCO and its $pD_2$ values were cromakalim 6.5, RP49356 6.3, pinacidil 5.92, nicorandil 4.43 and diazoxide 4.18. The relaxant effects of KCO were inhibited by glibenclamide (0.3, 1 and $10\;{\mu}M$) with $pA_2$ values of cromakalim 6.91, RP49356 6.59, pinacidil 6.55, nicorandil 5.97 and diazoxide 6.37. In addition, the relaxant effect of cromakalim or pinacidil was antagonised by TEA, a non-selective $K^+$ channel blocker, but not by apamin. Contractions induced by low concentration of KCI (< 40 mM) were inhibited by cromakalim $(100{\mu}M)$ and nicorandil $(300{\mu}M)$, but those evoked by higher concentration (> 40 mM) of KCI were little affected. In ovariectomized rat uterus, cromakalim dose-dependently inhibited oxytocin-induced contraction and glibenclamide $(10{\mu}M)$ inhibited the relaxant effect of cromakalim with $pD_2$ and $K_B$ values of 7.48 and $1.26{\times}10^{-7}M$, respectively. In estrogen-primed rat uterus, these values were 6.51 and $1.57{\times}10^{-7}M$, respectively, indicating that the cromakalim is less effective on the estrogen-treated uterine smooth muscle. Our results suggest that the KCO-sensitive $K^+$ channels participate in the motility of uterine smooth muscle and such channels are, at least in part, under the control of estrogen. In addition, our data Indicate that the type of $K^+$ channels activated by KCO is ATP-sensitive $K^+$ channels which is blocked by glibenclamide.

• IMMUNE NETWORK
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• v.5 no.2
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• pp.89-98
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• 2005

Background: DNA vaccination represents an anticipated approach for the control of numerous infectious diseases. Used alone, however, DNA vaccine is weak immunogen inferior to viral vectors. In recent, heterologous prime-boost vaccination leads DNA vaccines to practical reality. Methods: We assessed prime-boost immunization strategies with a DNA vaccine (minigene, $gB_{498-505}$ DNA) and recombinant vaccinia virus $(vvgB_{498-505})$ expressing epitope $gB_{498-505}$ (SSIEF ARL) of CD8+ T cells specific for glycoprotein B (gB) of herpes simplex virus (HSV). Animals were immunized primarily with $gB_{498-505}$ epitope-expressing DNA vaccine/recombinant vaccinia virus and boosted with alternative vaccine type expressing entire Ag. Results: In prime-boost protocols using vvgBw (recombinant vaccinia virus expressing entire Ag) and $vvgB_{498-505}$, CD8+ T cell-mediated immunity was induced maximally at both acute and memory stages if primed with vvgBw and boosted with $vvgB_{498-505}$ as evaluated by CTL activity, intracellular IFN-staining, and MHC class I tetramer staining. Similarly $gB_{498-505}$ DNA prime-gBw DNA (DNA vaccine expressing entire Ag) boost immunization elicited the strongest CD8+ T cell responses in protocols based on DNA vaccine. However, the level of CD8+ T cell-mediated immunity induced with prime-boost vaccination using DNA vaccine expressing epitope or entire Ag was inferior to those based on vvgBw and $vvgB_{498-505}$. Of particular interest CD8+ T cell-mediated immunity was optimally induced when $vvgB_{498-505}$ was used to prime and gB DNA was used as alternative boost. Especially CD7+ T cell responses induced by such protocol was longer lasted than other protocols. Conclusion: These facts direct to search for the effective strategy to induce optimal CD8+ T cell-mediated immunity against cancer and viral infection.

• Journal of Chest Surgery
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• v.9 no.2
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• pp.220-238
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• 1976

Six cases of congenital heart disease were operated on by means of cardiopulmonary bypass between December, 1975 and April, 1976. Two cases of ventricular septal defects (VSD), two cases of VSD, associated with ruptured aneurysm of sinus Valsalva, two cases of atrial septal defects (ASD) and one case of pulmonic stenosis with patent ductus arteriosus were operated. Sarns roller pumps and Bentley Temptrol oxygenators were used for extracorporeal circulation. Pump oxygenator was primed with Ringer's lactate solution, 5% dextrose in water, mannitol, and ACD blood. Flow rate ranged from 2.0 to $2.4L/M^2/min$. Bicarbonate was added to the oxygenator with estimated amount as 15 mEq/L/hr. Venous catheters were introduced into superior and inferior vena cava, and oxygenated blood was returned to the body through aortic cannula inserted into ascending aorta. Moderate hypothermia ($30^{\circ}C$) was induced by core cooling. Aorta was cross clamped for 15 minutes and released for 3 minutes, and repeated clamping when necessary. Atrial and ventricular septal efects were closed by direct sutures. Aneurysms of sinus Valsalva ruptured into the right ventricle were repaired through right ventriculotomy by d:rect closure with Dacron patch reinforcement. Cardiopulmonary bypass time varied from 66 to 209 minutes, and aorta cross clamping time ranged from 13 to 56 minutes. Postoperative bleeding was minimal except one case who needed for evacuation of substernal hematoma. Intra- and postoperative urinary output was satisfactory. Acid-base balance, partial pressure of $O_2$, electrolytes, and hematological changes during intra- and post-perfusion period remained at the acceptable ranges. No mortality was experienced.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.28 no.2
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• pp.209-218
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• 1995

Bacteriophage T7 gene 2.5 protein, a single-stranded DNA binding protein, has been implicated in T7 DNA replication, recombination, and repair. Purified gene 2.5 protein has been shown to interact with the phage encoded gene 5 protein (DNA polymerase) and gene 4 proteins (helicase and primase) and stimulates their activities. Genetic analysis of T7 phage defective in gene 2.5 shows that the gene 2.5 protein is essential for T7 DNA replication and growth. T7 phage that contain null mutants of gene 2.5 were constructed by homologous recombination. These mutant phage $(T7\Delta2.5)$ cannot grow in Escherichia coli. After infection of E. coli with $T7\Delta2.5$, host DNA synthesis is shut off, and $T7\Delta2.5$ DNA synthesis is reduced to less than $1\%$ of wild-type phage DNA synthesis (Kim and Richardson, 1993, Proc. Natl. Aca. Sci. USA, 90, 10173-10177). A truncated gene 2.5 protein $(GP2.5-\Delta21C)$ deleted the 21 carboxyl terminal amino acids was constructed by in vitro mutagenesis. $GP2.5-\Delta21C$ cannot substitute for wild-type gene 2.5 protein in vivo; the phage are not viable and exhibit less than $1\%$ of the DNA synthesis observed in wild-type phage-infected cells. $GP2.5-\Delta21C$ has been purified to apparent homogeneity from cells overexpressing its cloned gene. Purified $GP2.5-\Delta21C$ does not physically into「act with T1 gene 4 protein as measured by affinity chromatography and immunoblot analysis. The mutant protein cannot stimulate T7 gene 4 protein activity on RNA-primed DNA synthesis and primer synthesis. These results suggest that C-terminal domain of gene 2.5 protein is essential for protein-protein interactions.

• Journal of Embryo Transfer
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• v.10 no.1
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• pp.45-51
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• 1995

In order to determine the optimum condition and timing for in vitro maturation of oocytes to metaphase of meiosis II (M II), the immatured follicular oocytes were recovered by puncturing the large(1.0~1.5 mm in diameter) and small(<1.0 mm in diameter) follicles in the ovaries of rabbits treated intramuscularly with a single dose of 100 TU PMSG 68 hours previously. The follicular oocytes were classified into three grades by the attachment of cumulus cells. The Grade I and II follicular oocytes from large follicles were cultured in BO-DM medium with 10% FCS, 35 $\mu$g /nl of FSH, 10 $\mu$g /ml of LH and 1 $\mu$g /ml of estradiol-17$\beta$ at 39t in a 5% $CO_2$ incubator for 11 to 23 hours. In 3 hours interval during the culture period, the oocytes were harvested and their cumulus cells were removed with hyaluronidase. The denuded oocytes were stained with Hoechst 33342 dye and their meiotic status and extrusion of the first polar body (PB) were examined under a fluorescence microscope. Also the fragmentation of the first PB and the distance between the first PB and nucleus were examined. The results obtained were as follows: 1. The mean recovery rate of follicular oocytes from the large and small follicles was 59. 9 and 31.3%, respectively. The mean number of oocytes recovered per rabbit and the Grade I percentage were 14.6 and 94.4% in large follicles, but 2.1 and 61.1% in small follicles, respectively. All the parameters examined were different significantly (p<0.05) between both the folliclular size. 2. Most of the follicular oocytes(86.8%) were matured in vitro to M II phase in 14 hours in Grade I oocytes, but the significantly(p<0.05) less oocytes(45.5%) were matured in Grade II oocytes. 3. The first PB was extruded in most of the oocytes(94.7%) in 14 hours of culture with the fragmentation rate of 29.6%, but the fragmentation rate of the first PB increased significantly (p<0.05) as the culture period for maturation was longer to 20 hours(63.5%). 4. The distance between the first PB and nucleus was increased linearly (p<0.05) as the maturation time passed from 14(7.1$\mu$rn) to 23 hours(58.4$\mu$m). 5. From the above results it was concluded that the optimum time for in vitro maturation culture might be 14 hours in the follicular oocytes from rabbit primed with PMSG for 68 hours, expecially when these follicular oocytes were used for recipient cytoplasms in embryo cloning.

• Journal of Korean Neurosurgical Society
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• v.38 no.2
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• pp.126-131
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• 2005

Objective : The choice of tumor antigen for dendritic cell[DC]-loading has still been an unresolved problem in the DC-based vaccine strategies against malignant gliomas that has not been found well-characterized tumor specific antigens. In this study, we compare tumor-specific T cell response induced by glioma apoptotic body[GAB]-pulsed DCs to response induced by glioma cell lysate-pulsed ones quantitatively. Methods : DCs generated in the presence of granulocyte macrophage-colony stimulating factor and interleukin[IL]-4 from peripheral blood mononuclear cells[PBMCs] of HLA-A2 positive healthy donors were cultured. Each GABs and glioma cell lysate generated from HLA-A2 positive T98G glioblastoma cells were co-incubated with DCs. $CD8^+$ T lymphocytes isolated from PBMCs of same donors were cultured in media containing IL-2 and either stimulated by GAB- or lysate-pulsed DCs three times at a weekly interval. The interferon[IFN]-${\gamma}$ concentrations of each cell culture supernate were measured by enzyme immunoassay technique. Cytolytic activity of the generated cytotoxic $CD8^+$ T cells either stimulated with GAB- or lysate-pulsed DCs was determined by a standard 4-h $^{51}Cr$-release assay. Results : IFN-${\gamma}$ production and cytolytic activity of effector T cells stimulated by GAB-pulsed DCs were significantly higher than those of T cells stimulated by lysate-pulsed ones. Conclusion : These results indicate the choice of antigen is a critical determinant in the induction of antitumor immunity against malignant glioma. Antigen preparations from GABs represent a promising alternative to glioma cell lysate in DC-based glioma vaccine strategies.

• Korean Journal of Ichthyology
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• v.12 no.3
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• pp.173-179
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• 2000

Effects of Estradiol-$17\beta(E_2)$ and 2, 4-dichlorophenoxy acetic acid (2, 4-D) on vitellogenin(VTG) production were investigated in primary hepatocyte culture of olive flounder, Paralichthys olivaceus. Highest survival rate of hepatocyte were observed at $27^{\circ}C$, which markedly declined equal to 50% of those of $15^{\circ}C$. Vitellogenin production peaked at the concentration of $10^{-6}M\;E_2$. No effect was observed on VTG production at various concentrations of 2, 4-D. However, a low concentration of 2, 4-D (ie, $10^{-8}M$) only appeared increased VTG production. $E_2$ or $10^{-8}M$ 2, 4-D-primed VTG production was markedly inhibited by the addition of $10^{-6}M$ tamoxifen to the culture medium(P<0.01). Inhibition was not affected by combinational treatment with $10^{-6}M$ $E_2$ and $10^{-6}M$ 2, 4-D. These results from the current investigation suggest that 2, 4-D mimics $E_2$, but the mechanism of reaction in inducing the $E_2$ receptor are different in VTG production in oliver flounder hepatocytes.

• Journal of Bio-Environment Control
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• v.12 no.4
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• pp.221-227
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• 2003

The objective of this research was to determine the effect of seed priming on membrane integrity during poriming and germination. Among the five chemicals, $KNO_3$at 150 mL gave the shortest $T_{50}$ (days required to reach 50% of the final germination percentage). Compared to unprimed, the seeds primed with 150 mL $KNO_3$ at 20$^{\circ}C$ for 4 days had reduced $T_{50}$ values when germinated at 15$^{\circ}C$. These results indicated that seed priming is an effective way for rapid and synchronized germination, especially at low temperature. Changes in conductivity of priming solutions during the 4-days period of priming were highly dependent upon the priming agents. Conductivity of the $KNO_3$ and $K_3PO_4$ solution slowly declined during the first 3 hours ad then increased Amount of amino acids, sugars and proteins exuded from seeds into $KNO_3$ solution were less than those into distilled water and $K_3PO_4$. All the results suggested that the $KNO_3$ priming play a positive role in regulating the permeability of cell membranes.

• Journal of Bio-Environment Control
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• v.12 no.4
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• pp.228-234
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• 2003

Various hydration treatments have been devised to improve the rate and univormity of seed germination as well as seed viability. Hydropriming and priming, the imbibitaion of seeds in distilled water and osmotic solutions, respectively, are useful techniques for early establishment of uniform seedlings. In this study, the optimum conditions were determined for hysropriming of cucumber seeds. Effects of hydropriming were also compared to those of osmotic priming. Seeds, hydroprimed with distilled water and primed with 50 mM $NaNO_3$at 20 for 24 hour, showd shorter number of days required to reach 50% of the final germination percentage ($T_{50}$), and mean number of days to germinate (MDG) that those untreated seeds. Hydroprimed with 10 mL distilled water showed better results in improving the rate of germination thatn other treatments tested. Hydropriming of the aged seeds improved germination percentage at 15$^{\circ}C$ and 25$^{\circ}C$. The $T_{50}$ and MDG values were reduced significantly by hydropriming of the aged seeds regardless of germination temperature. In addition to being simple and inexpensive, the hydropriming resulted in early germination at low temperature. he hydropriming was also effective in increase of germination percentage and normal seedlings.

• Biomedical Science Letters
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• v.5 no.1
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• pp.85-93
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• 1999

Nitric oxide (NO) plays an important role in immunologic defense, and influences upon the functioning of secretory tissues and cells. It also exhibits cytotoxic/cytostatic activity as one of major operating effectors of the cellular immunity system. We investigated the effects of serum on the cell damages and NO production in the mouse liver cell line BNL CL.2 to establish the role of NO. We observed that, when BNL CL.2 cells were cultured in serum-free medium, they were induced to cell damage by the stimulation of IFN-$\gamma$ alone or IFN-$\gamma$ plus LPS. Serum-starved cells showed large amount of nitrite accumulation and NO synthase (NOS) expression in response to IFN-$\gamma$ alone in dose- and time- dependent manners, but serum-supplied cells did not The production of NO was blocked by protein tyrosine kinase (PTK) inhibitors, genistein and herbimycin. These results suggest that the deprivation of serum in the BNL CL.2 cell culture medium might primed with the cells to produce NO when the cells are triggered by IFN-$\gamma$ and the involvement of PTK signal transduction pathway in the expression of NOS gene in murine hepatocytes.