The objective of this study was to investigate the motility and kinematics of boar sperm that while stored at 4C. The samples of fresh boar semen were place into an extender, Androhep, and stored at $4^{\circ}C$. In three of these samples, cryoprotectants were added. The sperm's motilities and kinematics were evaluated by using microscope (${\times}400$) and the viability status was evaluated by using with eosin staining method. The 5 sample groups are; Goup A:Androhep (extender), stored at $17^{\circ}C$. Group B:Androhep (extender), stored at $4^{\circ}C$. Group C:Androhep (extender), + 3% glycerol (cryoprotectant), stored at $4^{\circ}C$. Group D:Androhep (extender), + 3% DMSO (cryoprotectant), stored at $4^{\circ}C$. Group E:Androhep (extender), + 3% ethylene glycol (cryoprotectant), stored at $4^{\circ}C$. In group A, the sperm's motility was reduced. On day one the sperm's motility was ($85.7{\pm}2.3$) and day 5 the motility was ($43.9{\pm}3.3$). In group B, C and D the sperm's motility were reduced to 0 on day 5. In group E the sperm's percentage of motility decreased. On day one the sperm's motility was ($42.0{\pm}0.5$) and day 5 the motility was ($2.3{\pm}0.3$). When comparing cryoprotectant in samples of boar sperm there is a slight improvement in the results when the use of Androhep Lite (extender), + 3% ethylene glycol (cryoprotectant), stored at $4^{\circ}C$ are used. Based on these results, ethylene glycol can protect sperm from heat shock at $4^{\circ}C$, but not satisfactory level. However, it showed the possibilities of liquid semen preservation at $4^{\circ}C$ by using cryoprotectant.
This study demonstrates the consolidation effect of ethylsilicate consolidants considering material characteristics and weathering degree of Samneunggyeseongakyukjonbul(rock-carved Yukjonbul Buddha in Samneung Valley) in Namsan, Gyeongju. The buddha statue is composed of alkali feldspar granite and contains numerous sets of joint with exfoliation and granular disintegration, therefore the statue is necessary to be treated for surface strength. The laboratory and in-situ tests of consolidation effect showed more increase of ultrasonic velocity that KSE 300, a relatively highly concentrated consolidant, performed more increase of ultrasonic velocity and decrease of porosity than others after treatments in weathered granite. And the consolidated rock with OH 100 was more resistant to salt weathering. For the buddha statues, KSE 300 is more applicable to enhance surface strength because it showed higher consolidation effect for long term than OH 100 and the statues has not been weathered by salts.
First. the purification and analysis of alliin in garlic from different origins by alliin-HPLC determination method were studied. Allinase in garlic was inactivated by heating in boiling water followed by extraction of alliin in garlic with 80% methanol. To remove free amino acids and alliin homologs in garlic, garlic extract was separated by cation exchange column which was packed with amberlite CG-120 resin using 40L d-water as eluent. Alliin in garlic extract was crystallized in a mixture of acetone (50$^{\circ}C$):H$_2$O:acetic acid=70:29:1 and then recrystallized in a mixture of acetone (50$^{\circ}C$):H$_2$O:acetic acid=75:24:1. Obtained alliin was identified by melting point. TLC, microscope observation and mass spectrometry. High performance liquid chromatography (HPLC) following pre-column derivatization of cystein derivatives with o-phthaldialdehyde/2-mercaptoethanol has succeessfully been applied to the analysis of various garlics. Each alliic of standard solution and garlic extract was derivatized to isoindole derivative by o-phthaldialdehyde /2-mercaptoethanol and then analyzed by HPLC. Six point calibration was done by using alliin peak area. Lineality was observed at 0 ∼ 1.0mg/ml of alliin concentration. Weighted regression line function was Y=6254X - 256077. By this function, alliin contents in various garlics were 0.34 ∼ 0.73% fresh weight. Second study was designed to evaluate the effects of garlic extracts of various concentrations on the growth of various pathogenes (Eubacterium limonsum, Bacteroides fragilis, Salmonella typhimurium, Salmonella typhi, Shigella sonnei, Kiebsiella pneumoniae, Enterobacter cloacae, Pserdomonas aeruginosa, Escherichia coli). For antimicrobial effects against microorganism, totally minimal inhibition concentrations (MIC) of alliin were from 5,000 to 20,000ppm. MIC of ethanol extract were 1,250 to 10,000ppm.
Kim, H.S.;Ryu, B.Y.;Oh, S.K.;Suh, C.S.;Kim, S.H.;Choi, Y.M.;Kim, J.G.;Moon, S.Y.;Lee, J.Y.
Clinical and Experimental Reproductive Medicine
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v.25
no.1
/
pp.59-64
/
1998
This study was designed to evaluate the influence of pronuclear age on the survival and post-thawing development after cryopreservation of mouse embryos. Freezing and thawing were performed in the different pronuclear stages of mouse embryos after IVF. Embryos were obtained from $F_1$ hybrid mice and classified into 4 groups according to the pronuclear stage (6hr, 9hr, 12hr and 15hr after insemination). Pronuclear ova were slowly cooled in a biological freezer using 1.5M 1,2-propanediol and 0.1M sucrose as cryoprotectant. Thawing was done at room temperature and 1,2-propanediol was removed by multi-step dilutions. Both frozen-thawed embryos and control fresh embryos were cultured in vitro in Ham's F-10 medium supplemented with 4mg/ml BSA. In control group, the development rate after 48hr was 99.3%, and the complete hatching rate after 144hr was 61.3%. In experimental groups, the survival rate after thawing was 95.4% in 6hr, 88.7% in 9hr, 75.2% in 12hr and 62.4% in 15hr after insemination, the development rate after 48hr was 61.1, 77.0, 67.0 and 79.6%, respectively, and the complete hatching rate after 144hr was 25.7, 43.7, 42.2 and 60.0%, respectively. The survival rate in 15hr was significantly lower (p<0.05) compared with other groups. In vitro development rates after 48hr were similar in all groups, but complement hatching rate was significantly lower (p<0.05) in 6hr group. In conclusion, cryopreservation of mouse pronuclear ova with 2 distinct pronuclei (9hr and 12hr groups) showed better results after thawing compared with early (6hr group) or late pronuclear ova just prior to cleavage (15hr group).
The present experiments on cryopreservation were designed to examine the effects of solution toxicity, equilibration time and cell stages on the post-thaw survival of bovine IVF embryos. The oocytes were matured in vitro(IVM) for 24 hrs. in TCM-199 supplemented with 35 $\mu$g /ml FSH, 10 $\mu$g /ml LH, 1 $\mu$g /ml estradiol-17$\beta$ and granulosa cells at 39$^{\circ}C$ under 5% $CO_2$ in air. They were fertilized in vitro(IVF) by epididymal spermatozoa treated with heparin for 24 hrs., and then the zygotes were co-cultured in vitro(IVC) with bovine oviductal epithelial cells for 7 to 9 days. The bovine IVF embryos were exposed to the EFS solution in one step at room temperature, kept in the EFS solution during different period for toxicity test, vitrified in liquid nitrogen, and thawed rapidly. 1. after the bovine blastocysts were exposed to EFS solution for 2 min. at room temperature and then they were washed in 0.5 M sucrose solution and TCM-199, they were cultured to examined cryoprotectant induced injury during exposure, Most of the embryos(95.0%) developed to reexpanded blastocoels. However, when the exposure time was extended to 5 and 10 min, these development rates dropped dramatically in 5 min. (69.5%) and 10 min. (47.4%), respectively, 2. When the bovine IVF embryos were vitrified in EFS solution after the equilibration for 1 and 2 min. exposure, The embryos to have reexpanded blastocoels following thawing, washing and culture processes were found to he 82.6 and 73.9%, respectively. However, when the exposure time was extended to 3 min, this survival rate dropped to 18.2%. The optimal time for equilibration of bovine IVF blastocysts in EFS solution seemed to he 1~2 min. 3. When the bovine IVF embryos were equilibrated for 1 min. the significantly (P<0. 05) higher post-thaw survival rates were obtained from the embryos of blastocyst stage(81.3%) than morulae stage(5. 1%). The optimal cell stage for viterification with EFS solution proven to he blastocyst stage in bovine IVF embryos. 4. The number of blastomeres of blastocyst stage was examined with nuclear staining with Hoechst 33342 during 7 to 9 days post-insemination. The cell counts of frozen bovine IVF embryos were found significantly(P$\geq$7.5 and those of the fresh embryos 76.6$\geq$7. 1, which were cultured in the sarne period and conditions as frozen embryos.
Cordyceps and its allies fungi has been described as a secret medicine that gives eternal youth and a long life. Some species of Cordyceps are sources of biochemicals, such ascordycepin, with interesting biological and pharmacological properties. Hence, it has been studied to uncover its pharmacological effect. We attempted to study the formation of fruiting bodies and to develop means of mass production Korean isolate of Paecilomyces tenuipes has been inoculated into silkworms, where it reproduced using culture container, mini-kit successfully. Culture container, mini-kit is composed of a cylinder-shaped body and lid. The container is made of translucent polyethylene terephthalate. The size of the container is $82{\times}75mm$, reduced by 10 times as compared with the conventional culture kit. The mini kit has many advantages - high culture amount, ability of maintaining optimal humidity, parasite-free cultivation and high-end appearance. With the kit, the optimal cultivation condition is under $22^{\circ}C$, culture period of 53 days. And synnemata of P. tenuipes could be kept fresh for 14 days at the temperature of under $10^{\circ}C$. Therefore, the Min-kit can be used in both ways as a culture container and a packing kit for end-user customers.
Go-Wun Yeo;Dong-Geun Lee;Ju-Hui Kim;Min-Joo Park;Jin Sun Kim;Yuck Yong Kim;Ki Hwan Yoo;Yong Jae Choi;Sang-Hyeon Lee
Journal of Life Science
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v.33
no.6
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pp.506-511
/
2023
The purposes of this study were to isolate and characterize lactic acid bacteria with antibacterial activity against Methicillin-resistant Staphylococcus aureus (MRSA) from fermented food and to confirm the antibacterial activities of fermented rice products using the isolated lactic acid bacterium. Three bacteria, namely, Weissella sp. ISF-1, ISF-2, and ISF-3, were selected from fermented squid based on the 16S rRNA gene sequence. All three strains grew well in an MRS medium containing 5% (w/v) NaCl and showed antibacterial activity against Bacillus cereus, Staphylococcus aureus, and MRSA. Their growth was excellent at 0% ~ 5% (w/v) NaCl and relatively good up to 7% (w/v) NaCl. The initial pH of 8 was optimal for their growth, and good growth was also observed at pH 6, 7, and 9. The lyophilisates of the fermented rice using Weissella sp. ISF-1 showed antibacterial activities against B. cereus, S. aureus, and MRSA. We inferred that isolated lactic acid bacteria could be useful in the development of probiotics and biopreservatives for foods and in the treatment of MRSA and may increase the value of rice products.
The mutant with high productivity, X. malvacearum SNUF 560-6, was acquired from the X. malvacearum SNUF 560 with low productivity by UV-light irradiation. It was preserved is lyophilized stock culture and it was transferred to PDA slant to maintain viability fortnightly. Fermentations were started by retransfering to MY agar slant from PDA stok culture. The experiments for optimal xanthan gum production were studied in a chemically defined medium. Of the carbon and nitrogen sources tested, 0.4% sucrose medium and 10mM glutamic acid medium yielded the highest xanthan gun production respectively. The addition of 10g/l succinic acid stimulated xanthan gum production. Also 65mM $PO_4\;^{-3}\;(12.6g/l\;KH_2PO_4)$ was effective on xanthan gum production. Finally, medium 1 and medium 2 which have high xanthan gum production potencies were achieved in this stud. The components of medium 1 and medium 2 were as follows: Medium 1 : sucrose 40g/l glutamate 10mM $PO_4\;^{-3}\;54mM\;(KH_2PO_4\;12.65g/l)$ Citrate 2g/l $MgSO_4{\cdot}7H_2O\;0.2g/l$$H_3BO_3\;0.005/l$ ZnO 0.006/l $FeCl_2{\cdot}6H_2O\;0.0024g/l$$CaCO_3\;0.02g/l$ Medium 2 : $Sucrose\;40g/l\;(NH_4)_2SO_4\;2g/l$$PO_4\;^{-3}\;65mM\;(KH_2PO_4\;12.65g/l)$ Succinate 10g/l $MgSO_4{\cdot}7H_2O\;0.02g/l$$H_3BO_3\;0.06g/l$ ZnO 0.006g/l $FeCl_2{\cdot}6H_2O\;0.0024g/l$$CaCO_3\;0.02g/l$.
Journal of the Korean Society of Food Science and Nutrition
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v.41
no.8
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pp.1041-1048
/
2012
Shiitake mushroom (SM; Lentinus edodes) are cultivated and consumed in many Asian countries including Vietnam, China, Japan, Korea, and Thailand. In Asia, SM are mainly dried and used as flavoring. The aim of this study was to compare the effects of SM created with different drying processes, such as oven-dried and sun-dried, on the antioxidative and antigenotoxic effects. Raw and dried SM were extracted with acetone, ethanol, methanol, and hot water. The antioxidant effects of SM were evaluated by determining total phenolic content, DPPH radical scavenging activity (RSA), an ORAC assay, and a cellular antioxidant capacity (CAC) assay. The inhibitory effect of SM on oxidative stress-induced DNA damage in human leukocytes was evaluated by a Comet assay. The total phenolic content of raw SM extracted with methanol and of that extracted with water were significantly higher than the dried SM. Among the water extracts, the $IC_{50}$ for DPPH RSA of raw and sun-dried SM were significantly higher than that of oven-dried SM. Sun-dried SM showed the most potent ORAC value at 50 g/mL. The CAC against $AAPH^-$ induced oxidative stress in HepG2 cells, and $H_2O_2$ induced DNA damage were effectively protected against by all SM extracts. These results suggest that unprocessed SM are the best antioxidants, and that the sun-dried method would be the best option to use in terms of antioxidant activity and the antigenotoxic effect.
Background : Defects in apoptotic signaling pathways play important role in tumor initiation, progression, metastasis and resistance to treatment. Several proteins which may promote tumorigenesis by inhibiting apoptosis were identified. The survivin protein is the member of inhibitor of apoptosis protein(IAPs) family which inhibits apoptosis. Unlike other IAPs, it is expressed in during the fetal period but not in adult differentiated tissues. Many reports have stated that survivin is selectively expressed in many cancer cell lines and cancer tissues. We performed immunohistochemical analysis for survivin expression in non-mall cell lung cancer to get evaluate its clinical implication. Methods : Twenty nine surgically resected lung cancers were examined. Immunohistochemical staining were performed by immuno-peroxidase technique using avidin-biotinylated horseradish pemxidase complex in the formalin-fixed, paraffin-embedded tissue $4{\mu}m$ section. Anti-survivin polyclonal antibody was used for primary antibody and anti-p53 monoclonal antibody was also used to analyze the correlation between survivin and p53 expression. The survivin expression scores were determined by as the sum of the stained area and intensity. Results : Immunohistochemical analysis showed cancer specific expression of survivin in 20 of 29 cases (69.0%). Western blot analysis also showed the selective survivin expression in tumor tissue. There was no correlation between survivin expression and clinicopathological parameters and prognosis. We analyzed the ∞π'elation between survivin expression and p53 expression, but found none. Conclusion: We confirmed the tumor specific expression of survival in non-small cell lung canær. But this expression was not correlated with clinical parameters as well as histology, tumor stage, recurrence, and survival rate. Also it was not statistically correlated with the expression of p53.
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