• Journal of Life Science
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• v.22 no.5
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• pp.575-581
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• 2012

This study was conducted to develop an economical optimum medium composition for the mass production of $Lactobacillus$ $plantarum$ and $Lactobacillus$ $reuteri$, livestock probiotics. Medium ingredient factors were selected on the basis of MRS broth composition, and the 15 ingredient variables were as follows: sucrose, glucose, molasses, yeast extract, corn steep liquor, soy peptone, dipotassium phosphate, manganese chloride, magnesium chloride, tween 80, sodium chloride, sodium acetate, ammonium citrate, sodium sulphate, and ferrous sulphate. The Plackett Burman design, consisting of 20 runs, was employed for the analysis of ingredient effects on cell growth of $L.$ $plantarum$ and $L.$ $reuteri$. As a result, sucrose, glucose, molasses, yeast extract, corn steep liquor, soy peptone, sodium acetate, and ammonium citrate positively influenced the growth of $L.$ $plantarum$. Additionally, yeast extract, soy peptone, $K_2PHO_4$, and tween 80 positively influenced the growth of $L.$ $reuteri$. Positive effects were found from sucrose, yeast extract, and soy peptone in the integrated analysis of the effects of both $L.$ $plantarum$ and $L.$ $reuteri$. Finally, effective medium components for both strains were found as follows: sucrose (20.0 g/l), glucose (5.0 g/l), soy peptone (11.0 g/l), yeast extract (5.0 g/l), $K_2PHO_4$ (0.2 g/l), $CH_3COONa$ (2 g/l), and $MgCl_2$ (0.02 g/l).

• Clinical and Experimental Reproductive Medicine
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• v.26 no.2
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• pp.137-148
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• 1999

Purpose of the present study was to find the optimal ovulation induction medicine for the maturation and development of immature oocytes and culture media for 2-cell embryos in the mouse model. ICR female mouse aged 6 to 8 weeks, were stimulated with 5 IU PMSG injection. At 47 to 50 hour post-PMSG injection, ovaries were dissected out and oocytes-cumulus complexes were punctured. The oocyte-cumulus complexes were cultured in media containing various ovulation induction medicine, CC, HMG and Metrodin for 18 hours. Female ICR mice were stimulated with 5 IU PMSG and 48 hours later were injected 5 IU of hCG, then female and male mice were mated. At 48 hour post-hCG injection, oviducts were dissected out and 2-cell embryos were flushed. The 2-cell embryos were cultured in various media, Ham's F-10 media of milli-Q water $(3^{\circ})$, Ham's F-10 media of HPLC (high performance liquid chromatography, Baxter) water, Medicult media, HTF (human tubal fluid) media for 96 hours. The results were as follows. 1. When the oocytes-cumulus complexes were cultured in $10^{-9}{\mu}g/ml{\sim}10^{-8}{\mu}g/ml$ of CC, those were suppressed in meiotic maturation $(28.2{\sim}33.7%)$. Whereas the oocytes-cumulus complexes were cultured in $10^{-7}{\mu}g/ml{\sim}10^{-4}{\mu}g/ml$, these were not effected in meiotic maturation $(54.5{\sim}72.7%)$. 2. When the oocytes-cumulus complexes were cultured in $10^{-4}{\mu}g/ml{\sim}10^{-1}{\mu}g/ml$ of Metrodin, those were suppressed in meiotic maturation $(35.7{\sim}41.5%)$. Meanwhile the oocytes-cumulus complexes were cultured in $10^{-7}{\mu}g/ml{\sim}10^{-5}{\mu}g/ml$, those were not effected in meiotic maturation $(54.2{\sim}70.3%)$. 3. When the oocytes-cumulus complexes were cultured in $10^{-5}{\mu}g/ml{\sim}10^{-4}{\mu}g/ml$ of HMG, those were suppressed in meiotic maturation $(48.2{\sim}50.4%)$. As being cultured in $10^{-7}{\mu}g/ml{\sim}10^{-6}{\mu}g/ml$, increased in meiotic maturation $(75.8{\sim}80.7%)$. 4. When the 2-cell embryos were cultured in Ham's F-10 media of milli-Q water $(3^{\circ})$, Ham's F-10 media of HPLC (high performance liquid chromatograpy, Baxter) water, Medicult media, HTF (human tubal fluid) media, developmental rates to blastocyst and hatching for 96 hour were 50.0%, 45.2%, 71.5% and 95.6%, respectively.

• The Journal of the Petrological Society of Korea
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• v.2 no.2
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• pp.74-94
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• 1993

Metabasites and metapelites in the Mungyong area were intruded by Cretaceous granites with radius of 4-8 km. As the distance from granite body increases, the mineral assemblage of metabasite changes from amphibole + plagioclase through amphibole + plagioclase + epidote to amphibole + plagioclase + epidote + chlorite. The compositional variations of amphibole and plagioclase according to the change of metamorphic grade and bulk rock compositions are very complex. Towards the Mungyong Cretaceous granite body, the mineral assemblage of metapelite changes from chlorite+ muscovite(ch1orite zone) through biotite + chlorite + muscovite(biotite zone) to andalusite+biotite + muscovite${\pm}$chlorite or cordierite+ biotite+ muscovite${\pm}$chlorite(cordierite zone). The estimated metamorphic conditions of cordierite zone are 480~$580^{\circ}C$ 1.5-3.3 kb. The theoretical study on the thermal metamorphism caused by the Cretaceous granite with radius longer than 4 km in the Mungyong area suggests the followings: The degree of metamorphism is mainly determined not by the size of granite body but by the temperature of granite intrusion; The country rocks within 2 km from Cretaceous granite have undergone metamorphism with temperature higher than $500^{\circ}C$, which is consistent with the petrological study in the Mungyong area. Mungyong Cretaceous granite caused a low P/T thermal metamorphism to the country rocks; the amphibolite facies metamorphism to the country rocks within 1-2 km from the granite body and the epidote-amphibolite and greenschist facies metamorphism to the country rocks within 2-5 km.

• Journal of The Korean Society of Clinical Toxicology
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• v.4 no.1
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• pp.7-16
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• 2006

Purpose: As basic information of antioxidant treatments for the patient with paraquat intoxication, in human peripheral lymphocytes, the cytotoxicity of paraquat was measured, and to evaluate the antioxidant effect of vitamin C and deferoxamine against this cytotoxicity, malondialdehyde (MDA), superoxide dismutase (SOD) activity and total antioxidant status (TAS) were measured. Methods: From 10 healthy adults, after obtaining a consent, 20ml peripheral blood was collected. Experimental groups were divided to (1) control group, the group treated with an identical amount of saline, (2) P group: the group treated with paraquat only, (3) PV group: the group treated with paraquat followed by vitamin C 30 minutes later, (4) PD group: the group treated with paraquat followed by deferoxamine 30 minutes later, (5) PVD group: the group treated with paraquat followed by vitamin C 30 minutes later and subsequently deferoxamine one hour later, and (6) PDV group: the group treated with paraquat followed by deferoxamine 30 minutes later and subsequently vitamin C 1 hour later, and thus to total 6 groups. In each group, 10 samples of peripheral blood was assigned and $100{\mu}M\;paraquat,\;100{\mu}M$ vitamin C, and $100{\mu}M$ deferoxamine were used as reagent. Lymphocytes were isolated, cultured, and cytotoxicity was measured by the Microculture Tetrazolium method (MTT assay), MDA and SOD activity, and TAS concentration were measured. Results: In regard to the cytotoxicity measured in each group, their cytotoxicity was decreased in the group treated with antioxidants, in comparison with the group treated with paraquat only. In the cases that the order of the treatment of these two antioxidants was altered, viability in the PDV group $(1.077{\pm}0.121)$ was increased more that the PVD group $(0.888{\pm}0.152)$ statistically significantly (p=0.018). Concerning the amount of MDA, in comparison with the P group $(6.78{\pm}0.93{\mu}mol/L)$, after the treatment of each antioxidant, the concentration of MDA was decreased statistically significantly (p<0.05). In the group treated with two antioxidants together, in comparison with the group treated only with one antioxidant, the amount of MDA was increased statistically significantly $(PV:\;3.96{\pm}0.98{\mu}mol/L,\;PD:\;4.92{\pm}1.50{\mu}mol/L,\;PVD:\;3.22{\pm}0.83{\mu}mol/L,\;and\;PDV:\;3.42{\pm}0.95{\mu}mol/L,\;p=0.007)$. The concentration of SOD measured in the blood in each group after the administration of paraquat, in comparison with the control group, a pattern of the elevation of SOD activity and subsequent decrease was detected, however, it was not statistically significant. In the comparison of the groups treated with antioxidants, in comparison with the P group $(1419.9{\pm}265.9{\mu}mol/L)$, SOD activity was decreased statistically significantly in only the PDV group $(1176.4{\pm}238.9{\mu}mol/L)$ (p=0.017). In regard to TAS measured in each group, in comparison with the P group $(0.87{\pm}0.05{\mu}mol/L)$, in all groups treated with the antioxidants, the PV group was $1.00{\pm}0.03{\mu}mol/L$ (p=0.005), the PD group was $9.01{\pm}0.24{\mu}mol/L$ was $4.64{\pm}3.98{\mu}mol/L$ (P=0.005), and the PDV group was $9.41{\pm}0.27{\mu}mol/La$ (p=0.005), and thus total antioxidant activity was increased statistically significantly In a multiple comparison test, the PDV group showed the highest total antioxidant activity (p<0.0001). Conclusion: The result of the assessment of the antioxidant effect of vitamin C and deferoxamine on paraquat-induced cytotoxicity showed that in regard to cytotoxicity, SOD activity and TAS measurement, the best result was observed in the PDV group. Therefore, it was found that vitamin C and deferoxamine were effective antioxidants for the paraquat-induced cytotoxicity, and it suggests that the administration of deferoxamine followed by vitamin C may improve their antioxidant effect more.

• Journal of Chest Surgery
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• v.34 no.9
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• pp.698-703
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• 2001

Background: Endovascular stent-graft insertion in aortic diseases is now generally accepted as an attractive alternative treatment modality. We reviewed our clinical experiences of endovascular stent-graft insertion in thoracic aorta. Material and Method: Since 1995, we performed 8 cases of endovasclar stent-graft insertion. Preoperative diagnoses were aortic aneurysms in 4, traumatic aortic ruptures in 3, and ruptured aortic pseudoaneurysm in 1. All procedures were performed in angiography room with the guidance of fluoroscopy. The stent-graft device is a custom-made 0.35mm thickness Z-shaped stainless steel wires, intertwined with each other using polypropylene suture ligation. It is covered with expanded Dacron vascular graft. Result: All procedures were performed successfully. Follow-up studies revealed 2 minimal perigraft leakages. There was no significant leakage or graft migration. 2 patients expired due to multiple organ failure and fungal sepsis. Other survivors(6) are doing well. Conclusion: Endovascular stent-graft insertion is relatively saft and effective treatment modality in the managment of various types of aortic diseases. In may be an effective alternative in aortic diseases of great surgical risk.

• Journal of Periodontal and Implant Science
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• v.26 no.1
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• pp.1-26
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• 1996

This study was performed to estimate the effects of cultured bone cell inoculated on porous type hydroxyaptite for the regeneration of the artificial alveolar bone defect. In this experiment 3 beagle dogs were used, and each of them were divided into right and left mandible. Every surgical intervention were performed under the general anesthesia by using with intravenous injection of Pentobarbital sodium(30mg/Kg). To reduce the gingival bleeding during surgery, operative site was injected with Lidocaine hydrochloride(l:80,000 Epinephrine) as local anesthesia. After surgery experimental animal were feeded with soft dietl Mighty dog, Frisies Co., U.S.A.) for 1 weeks to avoid irritaion to soft tissue by food. 2 months before surgery both side of mandibular 1st premolar were extracted and bone chips from mandibular body were obtained from all animals. Bone cells were cultured from bone chips obtained from mandible with Dulbecco's Modified Essential Medium contained with 10% Fetal Bovine Serum under the conventional conditions. Porous type hydroxyapatite were immerse into the high concentrated cell suspension solution, and put 4 hours for attachin the cells on the surface of hydroxyapatite. Graft material were inserted on the artificial bone defect after 3 days of culture. Before insertion of cellinoculated graft material, scanning electronic microscopic observation were performed to confirm the attachment and spreading of cell on the hydroxyapatite surface. 3 artificial bone defects were made with bone trephine drill on the both side of mandible of the experimental animal. First defect was designed without insertion of graft material as negative control, second was filled with porous replamineform hydroxyapatite inoculated with cultured bone marrow cells as expermiental site, and third was filled with graft materials only as positive control. The size of every artificial bone defect was 3mm in diameter and 3mm in depth. After the every surgical intervention of animals, oral hygiene program were performed with 1.0% chlorhexidine digluconate. All of the animals were sacrificed at 2, 4, 6 weeks after surgery. For obtaining histological section, tissus were fixed in 10% Buffered formalin and decalcified with Planko - Rycho Solution for 72hr. Tissue embeding was performed in paraffin and cut parallel to the surface of mandibular body. Section in 8um thickness of tissue was done and stained with Hematoxylin - Eosin. All the specimens were observed under the light microscopy. The following results were obtained : 1. In the case of control site which has no graft material, less inflammatory cell infiltration and rapid new bone forming tendency were revealed compared with experimental groups. But bone surface were observed depression pattern on defect area because of soft tissue invasion into the artificial bone defect during the experimental period. 2. In the porous hydroxyapatite only group, inflammatory cell infiltration was prominet and dense connective tissue were encapsulated around grafted materials. osteoblastic activity in the early stage after surgery was low to compared with grafted with bone cells. 3. In the case of porous hydroxyapatite inoculated with bone cell, less inflammatory cell infiltration and rapid new bone formation activity was revealed than hydroxyapatite only group. Active new bone formation were observed in the early stage of control group. 4. The origin of new bone forming was revealed not from the center of defected area but from the surface of preexisting bony wall on every specimen. 5. In this experiment, osteoclastic cell was not found around grafted materials, and fibrovascular invasion into regions with no noticeable foreign body reaction. Conclusively, the cultured bone cell inoculated onto the porous hydroxyapatite may have an important role of regeneration of artificial bone defects of alveolar bone.

• Journal of the korean academy of Pediatric Dentistry
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• v.30 no.2
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• pp.217-228
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• 2003

Co-ordinate growth of the brain and skull is achieved through a series of tissue interactions between the developing brain, the growing bones of the skull and the sutures that unite the bones. Craniosynostosis, the premature fusion of cranial sutures, presumably involves disturbance of these interactions. Bmp2, one of bone morphogenetic proteins (Bmps), is involved in the regulation of the shapes of individual bones and the relative proportions of the skeleton. Mutations in the homeobox gene Msx2, known as a downstream gene of Bmp, cause Boston-type human craniosynostosis. The phenotype of Dlx5 homozygote mutant mouse presents craniofacial abnormalities including a delayed ossification of calvarial bone. These facts suggest important roles of Bmp2, Msx2 and Dlx5 genes in the cranial bone growth and suture morphogenesis. To elucidate the function of these molecules in the early morphogenesis of mouse cranial sutures, we first analyzed by in situ hybridization the expression of Bmp2(E15-18), Msx2 and Dlx5 genes in the developing sagittal suture of calvaria during the embryonic stage. Bmp2 mRNA was intensely expressed in the osteogenic fronts and also at the low level in the periosteum of parietal bones during embryonic stage, Msx2 mRNA was intensely expressed in the sutural mesenchyme and mildly expressed in the dura mater during the embryonic stage. Dlx5 mRNA was intensely expressed osteogenic fronts and parietal bones. To further examine the role of Bmp signaling in cranial suture, we did in vitro experiments in E15.5 mouse calvarial explants. Interestingly, implantation of Bmp2-soaked beads onto the osteogenic fronts after 48 hours organ culture resulted in the increase of the tissue thickness and cell number around Bmp2 beads, compared to BSA control beads. In addition Bmp2 induced etopic expressions of Msx2 and Dlx5 genes. On the other hand, overexpression of FGF2 did not induce the expression of Msx2 and Dlx5. Taken together, these data indicate that Bmp2 signaling molecule has a important role in regulating the cranial bone growth and early morphogenesis of cranial suture. We also suggest that Bmp signaling is involved in all the stages of osteogenesis of cranial bones and the maintenance of cranial suture by regulating Msx2 and Dlx5 genes, and that Msx2 and Dlx5 genes are specific transcription factors of Bmp signaling pathway.

• Journal of the Korean Society of Food Science and Nutrition
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• v.31 no.6
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• pp.1084-1091
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• 2002

Mortierella alpina, a common soil fungus, is the most efficient organism for production of production acid presently known. Since arachidonic acid are important in human brain and retina development, it was undertaken the growing effect containing diet as a food ingredient. Arachidonic acid rich oil derived from Mortierella alpina, was subjected to a program of studies to establish for use in diet supplement. This study was compared the growth and learning effect of fungal oil rich in arachidonic acid by incorporated into diets ad libitum. Sprague-Dawley rats received experimental diets 5 groups (standard AIN 93 based control with beef tallow, extract oil 8%, and 4%, and Mortierella alpina in diet 10% and 20%) over all experiment duration (pre-mating, mating, gestation, lactation, and after weaning 4 weeks). Pups born during this period consumed same diets after wean for 4 weeks. There was no statistical significance of diet effects in reproductive performance and fertility from birth to weaning. But the groups of Mortierella alpine diet were lower of weight gain and diet intake after weaning. The serum lipids were significantly different with diet groups, higher TG in LO (oil 4%) group of dams, and higher total cholesterol in LF (M. alpina 10%) of pups, although serum albumin content was not significantly different in diet group. The spent-time and memory effect within 4 weeks of T-Morris water maze pass test in dam and 7-week- age pups did not differ in diet groups. On the count of backing error in weaning period of pups was lower in HO(extracted oil 8%) group. In the group of 10% and 20% Mortierella alpina diet, DNA content was lower in brain with lower body weight, but liver DNA relative to body weight was higher than control. Further correlation analyses would be needed DNA and arachidonic acid intakes, with Mortierella alpina diet digestion rate.

• Korean Journal of Environmental Biology
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• v.18 no.3
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• pp.331-336
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• 2000

In order to examine the effect of SO$_2$, which is the major component of acid rain, on the peroxidase activity, rice (Oryza sativa) seedlings were grown on the media containing various concentrations of Na$_2$SO$_3$. Na$_2$SO$_3$ concentrations needed for the 50% inhibition of rice seed germination were determined to be 300$\mu\textrm{g}$/ml at pH 7, 8$\mu\textrm{g}$/ml at pH 5 and 2$\mu\textrm{g}$/ml at pH 3. Notably, about 8 fold and 4 fold increase of the specific activity of the enzyme were observed with the seedlings treated with 8$\mu\textrm{g}$/ml Na$_2$SO$_3$ at pH 5 and 2$\mu\textrm{g}$/ml Na$_2$SO$_3$ at pH 3, respectively. The effects of Cd and Pb on the peroxidase activities and chlorophyll contents were also examined. About 3.9 fold higher peroxidase activities were found at 0.03mM Cd, and the chlorophyll contents were reduced to 63% of the control seedlings. At 0.04mM Pb, 2.5 fold higher enzyme activities were found and the chlorophyll contents were reduced to 72%. Therefore, the increases of rice peroxidase activities might be involved in the defense mechanism of the cell against various environmental stresses such as Na$_2$SO$_3$, Cd and Pb. The effects of Cu and Fe, which are the inducers of oxidative stresses by the generations of reactive oxygen species, on the peroxidase activities were also investigated. About 57% and 65% activity losses were found at 0.5mM CuSO$_4$ and 0.5mM FeSO$_4$, respectively, and radical scavenger ethanol almost completely protected both inactivations. However, dimethyl sulfoxide, mannitol, thiourea and histidine showed different radical scavenging effects one another against Cu and Fe inactivation.

• Journal of Life Science
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• v.26 no.12
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• pp.1376-1382
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• 2016

Xylitol is widely used in the food and medical industry. It is produced by the reduction of xylose (lignocellulosic biomass) in the Saccharomyces cerevisiae strain, which is considered genetically safe. In this study, the expression system of the GRE3 (YHR104W) gene that encodes xylose reductase was constructed to efficiently produce xylitol in the S. cerevisiae strain, and the secretory production of xylose reductase was investigated. To select a suitable promoter for the expression of the GRE3 gene, pGMF-GRE3 and pAMF-GRE3 plasmid with GAL10 promoter and ADH1 promoter, respectively, were constructed. The mating factor ${\alpha}$ ($MF{\alpha}$) signal sequence was also connected to each promoter for secretory production. Each plasmid was transformed into S. cerevisiae $SEY2102{\Delta}trp1$, and $SEY2102{\Delta}trp1$/pGMF- GRE3 and $SEY2102{\Delta}trp1$/pAMF-GRE3 transformants were selected. In the $SEY2102{\Delta}trp1$/pGMF-GRE3 strain, the total activity of xylose reductase reached 0.34 unit/mg-protein when NADPH was used as a cofactor; this activity was 1.5 fold higher than that in $SEY2102{\Delta}trp1$/pAMF-GRE3 with ADH1 as the promoter. The secretion efficiency was 91% in both strains, indicating that most of the recombinant xylose reductase was efficiently secreted in the extracellular fraction. In a baffled flask culture of the $SEY2102{\Delta}trp1$/pGMF-GRE3 strain, 12.1 g/l of xylitol was produced from 20 g/l of xylose, and ~83% of the consumed xylose was reduced to xylitol.