• Journal of Embryo Transfer
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• v.28 no.3
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• pp.265-271
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• 2013

Cryopreservation and in vitro fertilization (IVF) protocols are important in genetic studies and applications to transgenic animals. Various studies about boar sperm cryopreservation have been studied for a long time. Those were about the use of extenders, the choice of sugars, the cooling and warming rates. The factors that influence the boar sperm are the dramatic changes in temperatures, osmotic and toxic stresses, and reactive oxygen species (ROS) generation. Among these factors, ROS generation is the main damage to DNA which is a principal genetic material and the most important for the practical applications. So we wondered whether ROS generation could be reduced. In previous study, monothioglycerol (MTG) was essential for the culture of embryo stem cells. Therefore we added MTG in the freezing extender based on lactose-egg yolk (LEY) with trehalose. For the assessment of the frozen-thawed sperm, we focused onmotility, membrane integrity and DNA damage. First, we used a computer-aided sperm analysis system for overall conditions of sperm such as motility and viability. Then we performed the sperm chromatin structure assay for DNA integrity and hypo-osmotic swelling test for membrane integrity. And our result showed the existence of MTG in the freezing extender caused less damage to DNA and higher motility in frozen-thawed boar sperm. Also we checked a relative antioxidant activity of MTG in modified Modena B extender. We concluded that this reagent can activate sperm mitochondria at MTG $0.2{\mu}M$, contribute to sperm motility and DNA integrity but there was no significant difference on membrane integrity. Also antioxidant activity of MTG in modified Modena B extender was proved.

• Journal of Microbiology and Biotechnology
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• v.7 no.2
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• pp.114-120
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• 1997

Microorganisms producing xylanase were screened for the enzymatic production of xylo-oligo saccharides from xylan. One of the bacteria isolated from compost produced an endoxylanase extracellularly. The bacterium was identified as Bacillus sp. according to its taxonomic characteristics examined. Xylanase production reached upto 5 U/ml after 22 h of culture in LB medium at $30^{\circ}C$. The xylanase was purified by ammonium sulfate precipitation and gel filtration. The molecular weight of the xylanase was estimated to be 20,400 by SDS-PAGE. Optimal temperature and pH for the xylanase activity was $60^{\circ}C$ and 6.5, respectively. The enzyme was stable at temperatures upto $40^{\circ}C$ and pH values from 4 to 10. The xylanase was completely inhibited by the addition of 2 mM mercury ion. Apparent $K_m$ and $V_max$ values for oat spelt xylan were 9.2 mg/ml and 1954 U/mg protein, respectively. For birchwood xylan, the values were 6.3 mg/ml and 1009 U/mg protein. The predominant products of the xylan hydrolysis were xylobiose, xylotriose and xylotetraose, indicating that the enzyme is an endoxylanase. Upto $85{\%}$ of the initially added enzyme (2 U/ml) was bound to 50 mg/ml of the insoluble fraction of oat spelt xylan after incubation at $30^{\circ}C$ for 30 min.

• Fisheries and Aquatic Sciences
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• v.17 no.3
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• pp.345-353
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• 2014

Copper has been widely used to control algae and pathogens in fish culture ponds. However, its toxic effects on fish depend not only on its concentration in the water but also on the water quality. A laboratory experiment was conducted to assess copper toxicity in the black rockfish Sebastes schlegeli using a panel of antioxidant enzymes, including glutathione (GSH), glutathione S-transferase (GST), glutathione peroxidase (GPx), glutathione reductase (GR) and superoxide dismutase (SOD), at different levels of copper at three water temperatures (WT, 18, 23, $28^{\circ}C$) for 4 days. After exposure to two copper concentrations (100 and $200{\mu}g/L$), GSH levels and GST activities increased significantly, depending on WT (P < 0.05) in the liver, gill, and kidney of the black rockfish. GPx and SOD activities decreased significantly with both increasing WT and copper treatment in the organs of black rockfish (P < 0.05). These changes can be seen as initial responses to temperature stress and as a sustained response to copper exposure. This also indicates that GSH and related enzymes activities were sensitive indexes to stress by toxicants such as copper. The present findings suggest that simultaneous stress due to temperature change and copper exposure can accelerate changes in enzymes activities in the black rockfish. This provides another example of synergism between environmental temperature and pollutants, which may have important implications for the survival of fish in polluted environments during seasonal warming and/or global climate change.

• Journal of Microbiology and Biotechnology
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• v.20 no.3
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• pp.615-621
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• 2010

The viability of low-temperature sulfate reduction with hydrogen as electron donor was studied with a bench-scale gas-lift bioreactor (GLB) operated at $9^{\circ}C$. Prior to the GLB experiment, the temperature range of sulfate reduction of the inoculum was assayed. The results of the temperature gradient assay indicated that the inoculum was a psychrotolerant mesophilic enrichment culture that had an optimal temperature for sulfate reduction of $31^{\circ}C$, and minimum and maximum temperatures of $7^{\circ}C$ and $41^{\circ}C$, respectively. In the GLB experiment at $9^{\circ}C$, a sulfate reduction rate of 500-600 mg $l^{-1}d^{-1}$, corresponding to a specific activity of 173 mg ${SO_4}^{2-}g\;VSS^{-1}d^{-1}$, was obtained. The electron flow from the consumed $H_2$-gas to sulfate reduction varied between 27% and 52%, whereas the electron flow to acetate production decreased steadily from 15% to 5%. No methane was produced. Acetate was produced from $CO_2$ and $H_2$ by homoacetogenic bacteria. Acetate supported the growth of some heterotrophic sulfate-reducing bacteria. The sulfate reduction rate in the GLB was limited by the slow biomass growth rate at $9^{\circ}C$ and low biomass retention in the reactor. Nevertheless, this study demonstrated the potential sulfate reduction rate of psychrotolerant sulfate-reducing mesophiles at suboptimal temperature.

• Journal of Plant Biotechnology
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• v.30 no.1
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• pp.41-45
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• 2003

A study was under taken to investigate the appropriate explant sources of flower organ and suitable cultural conditions such as light, temperature, and sucrose in plant regeneration of Iris ensata culture. Explants of perianth, ovary, pedicel, and peduncle of Iris ensata were cultured at different daylength (0, 8, 16, 24 hour), different temperatures (10, 15, 25, 3$0^{\circ}C$), and sucrose concentrations (1, 3, 6, 9%) on MS medium. Formation of adventitious roots from explants of Iris ensata was effective in the dark, while that of adventitous shoots was effective in the light. The optimum daylength for young plant regeneration was 16 hours. The optimum temperature for shoot formation of Iris ensata explants was $25^{\circ}C$ but the formation at 10 and 15$^{\circ}C$ was ineffective. Especi-ally, perianth and ovary was effective in shoot formation from flower organ expants. T-he optimum concentration of sucrose for shoots and roots formation of Iris ensata explants was 3 and 6%, respectively.

• Korean Journal of Veterinary Research
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• v.33 no.2
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• pp.277-283
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• 1993

In the present study, observations were made on the life-history of Lymnaea viridis under laboratory conditions, involving incubation period of the eggs and their hatching rate, shell length of the newly hatched snails, sexual maturity, size of the snails when the snail produced the first egg-mass, the number of eggs in each egg-mass, egg-laying, ovipostion, growth rate of the snails, and longevity of the snail. At temperatures between $19.8^{\circ}C$ to $22.5^{\circ}C$, incubation period of the eggs occupied 10~12 days, and after beginning of hatching, all young snails emerged completely from the egg-mass within 5 days. The hatching rate was 88%. The average shell length of the newly hatched snails was about 0.064cm. The rate of growth was extraordinarily rapid under good laboratory conditions. When two snails were reared in one culture vessel($20{\times}15{\times}5cm$) with blue-green algae at about $22^{\circ}C$, snail growth was optimal, taking 37 days to reach 1.2cm in shell length. Sexual maturity reached in about 19 days. The size of the snails at sexual maturity was $0.78{\pm}0.05cm$ in length and $0.47{\pm}0.04cm$ in width. The first egg-masses produced were $0.59{\pm}0.22cm$ in length and $0.34{\pm}0.08cm$ in width, and contained 7~38 eggs. The eggs are usually laid in water. The egg-laying was affected by food and temperature. Snails fed with blue-green algae at about $22^{\circ}C$ produced larger egg-masses than the snails fed with fish food at about $26^{\circ}C$. Under conditions of continuous activity and growth, the maximum expectation of life appears to be 109~350(mean 230) days. And the shell length of snails at death were 1.39~1.64cm.

• Journal of Animal Science and Technology
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• v.51 no.6
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• pp.527-536
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• 2009

A potent protein degrading bacterium was isolated from soil samples of different environments. Polyphasic taxonomic studies and phylogenetic 16S rRNA sequence analyses led to identify the isolate IB No. 11 as a strain of Bacillus subtilis. The isolated strain was recognized to produce protease constitutively, and the maximum production (1.64 units/ml) was attained in a shake flask culture when the isolate was grown at $40^{\circ}C$, for 32 h in basal medium supplemented with starch (0.25%) and gelatin (1.25%) as sole carbon and nitrogen source, respectively. The optimum pH and temperature for the protease activity were determined to be pH 7.0 and $50^{\circ}C$, respectively. $Ca^{2+}$ and $Mn^{2+}$ enhanced remarkably the protease activity but neither showed positive effect on the protease's thermal stability. In addition, it was observed that the protease was fairly stable in the pH range of 6.5-8.0 and at temperatures below $50^{\circ}C$, and it could be a good candidate for an animal feed additive. The inhibition profile of the protease by various inhibitors indicated that the enzyme is a member of serine-proteases. A combination of UV irradiation and NTG mutagenesis allowed to develop a protease hyper-producing mutant strain coded as IB No. 11-4. This mutant strain produced approximately 3.23-fold higher protease activity (6.74 units/mg) than the parent strain IB No. 11 when grown at $40^{\circ}C$ for 32h in the production medium. The protease production profile of the selected mutants was also confirmed by the zymography analysis.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2010.10a
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• pp.15-15
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• 2010

Ginseng contained many different kinds of saponin which was the most valuable for people, but its yield cannot satisfy the demand using traditional extract methods. Enzyme transformation is a conformable and highly performed method which was fit for today. A ${\beta}$-glucosidase producing bacterium ($DCY51^T$) was isolated from Korean fermented-vegetable food kimchi. The 16S rRNA gene sequence analysis revealed that the strain $DCY51^T$ belongs to the genus Lactobacillus. The highest sequence similarity was found with Lactobacillus paracollinoides LMG $22473^T$ and Lactobacillus collinoides LMG $9194^T$ with levels of 16S rDNA similarity of 97.4% and 97.3%, respectively. Based on the above results the strain $DCY51^T$ placed in the genus Lactobacillus and proposed a new species, Lactobacillus kimchicus sp. nov. $DCY51^T$ (= KCTC $12976^T$ = JCM $15530^T$). It was culture solution reacted with Red Ginseng extract and $Rb_1$, respectively. The medium of bacteria was the liquid of MRS, the temperatures of growing and reacting between bacteria liquid and saponin were samely $37^{\circ}C$, there spective reacting time were 12 hours and 48 hours. Thus we got different saponins, and TLC and HPLC analysis showed that: enzyme respectively reacted with $Rb_1$ and Red Ginseng extract got the transformed saponin, respectively. The polarity position in TLC was a little higher than Rd; and the polarity position was the same as that of Compound K's, the saponin obtained from HPLC and other experimental results was not Compound K. The constitution of its saponin was hoped to be further confirmed.

• Korean Journal of Medicinal Crop Science
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• v.7 no.1
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• pp.7-10
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• 1999

The effects of storage temperature and periods as pretreatments on direct somatic embryogenesis or shoot development in liquid culture of leaves, stems and petioles of Rehmannia glutinosa and Scrophularia buergeriana were investigated. Proper storage Temperature for Rehmannia glutinosa was $8^{\circ}C$ and the leaf stock could be stored up to 12 weeks. In Scrophularia buergerjana. shoots were developed from the 20 week-stored petiole and stem at $8^{\circ}C$. In general, $8^{\circ}C$ was considered the most effective storage temperature in terms of storage cost.

• Journal of Ginseng Research
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• v.41 no.4
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• pp.463-468
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• 2017

Background: Both Panax ginseng Meyer and Panax quinquefolius are obligate shade-loving plants whose natural habitats are broadleaved forests of Eastern Asia and North America. Panax species are easily damaged by photoinhibition when they are exposed to high temperatures or insufficient shade. In this study, a cytohistological study of the leaf structures of two of the most well-known Panax species was performed to better understand the physiological processes that limit photosynthesis. Methods: Leaves of ginseng plants grown in soil and hydroponic culture were sectioned for analysis. Leaf structures of both Panax species were observed using a light microscope, scanning electron microscope, and transmission electron microscope. Results: The mesostructure of both P. ginseng and P. quinquefolius frequently had one layer of non-cylindrical palisade cells and three or four layers of spongy parenchymal cells. P. quinquefolius contained a similar number of stomata in the abaxial leaf surface but more tightly appressed enlarged grana stacks than P. ginseng contained. The adaxial surface of the epidermis in P. quinquefolius showed cuticle ridges with a pattern similar to that of P. ginseng. Conclusion: The anatomical leaf structure of both P. ginseng and P. quinquefolius shows that they are typical shade-loving sciophytes. Slight differences in chloroplast structure suggests that the two different species can be authenticated using transmission electron microscopy images, and light-resistant cultivar breeding can be performed via controlling photosynthesis efficiency.