• Journal of Embryo Transfer
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• v.25 no.3
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• pp.189-193
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• 2010

We investigated the cleavage rate and blastocyst yield for each culture condition to enhance tolerance of cryo-preservation of bovine IVF embryo with relatively lower cryo-tolerance compared to in vivo embryo. The cleavage rate and blastocysts yield for CR1aa, IVMD, IVD, CR1aa+10% FBS were 73.2, 69.3, 72.8, 68.5% and 44.1, 30.8, 33.3, 48.0%, respectively. The values did not differ among each treatments without serum. For embryo vitrification, In vivo and In vitro blastocysts were exposed to VS1(10% glycerin, 0.1 M glucose, 0.1 M sucrose, PEG 1%) for 5 min, and VS2 (10% glycerin, 10% EG, 0.2 M glucose, 0.2 M sucrose, PEG 2%) for 5 min and then VS3 (10% glycerin, 30% EG, 0.3 M glucose, 0.3 M sucrose, PEG 3%) for 1 min. The exposed embryos were then loaded into the 0.25 ml plastic straws and then plunged into liquid nitrogen. The straws were held for period of 1 to 2 weeks before thawing. In embryo viability, no differences in blastocyst re-expansion rates were found between in vivo and in vitro embryos. whereas expansion-BL rates was significantly higher for in vivo-derived embryos (72.7%) when compared to in vitro-derived embryos (51.4%), respectively (P<0.05). In conclusion, our results indicate that combined use of CRIaa culture medium with vitrification might enhance tolerance of cryopreservation for bovine IVF embryo production.

• Toxicological Research
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• v.24 no.1
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• pp.69-78
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• 2008

Mutant mouse which show dwarfism has been developed by N-ethyl-N-nitrosourea (ENU) mutagenesis using BALB/c mice. The mutant mouse was inherited as autosomal recessive trait and named Small Body Size (SBS) mouse. The phenotype of SBS mouse was not apparent at birth, but it was possible to distinguish mutant phenotype from normal mice 1 week after birth. In this study, we examined body weight changes and bone mineral density (BMD), and we also carried out genetic linkage analysis to map the causative gene(s) of SBS mouse. Body weight changes were observed from birth to 14 weeks of age in both affected (n = 30) and normal mice (n = 24). BMD was examined in each five SBS and normal mice between 3 and 6 weeks of age, respectively. For the linkage analysis, we produced backcross progeny [(SBS${\times}$C57BL/6J) $F_1{\times}$ SBS] $N_2$ mice (n = 142), and seventy-four microsatellite markers were used for primary linkage analysis. Body weight of affected mice was consistently lower than that of the normal mice, and was 43.7% less than that of normal mice at 3 weeks of age (P < 0.001). As compared with normal mice at 3 and 6 weeks of age, BMD of the SBS mice was significantly low. The results showed 15.5% and 14.1 % lower in total body BMD, 15.3% and 8.7% lower in forearm BMD, and 29.7% and 20.1% lower in femur BMD, respectively. The causative gene was mapped on chromosome 10. The map order and the distance between markers were D10Mit248 - 2.1 cM - D10Mit51 - 4.2 cM - sbs - 0.7 cM - D10Mit283 - 1.4cM - D10Mit106 - 11.2cM - D10Mit170.

• Proceedings of the KSAR Conference
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• 2003.06a
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• pp.90-90
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• 2003

Circling mice were recorded to display profound deafness and a head-tossing and bidirectional circling behavior, showing an autosomal recessive mode of inheritance. In addition, the histological examination of inner ears revealed that the region around organ of Corti, spiral ganglion neurons and outer hair cells showed definite abnormality. On the other hand, a genetic linkage map was constructed in an intraspecific backcross between cir and C57BL/6J mice. The cir gene was mapped to a region between D9Mitl16/D9Mit15 and D9Mit38 on the mouse chromosome 9. Estimated distances between cir and D9Mitl16, and between cir and D9Mit38 are 0.70 $\pm$ 0.40 and 0.23 $\pm$ 0.23 cM, respectively. The markers in order was defined as follows: centromere-D9Mit182- D9Mit51/ D9Mit79/ D9Mit310- D9Mit212/ D9Mit184- D9Mit116/ D9Mit15- cir- D9Mit38- D9Mit20- D9Mit243- D9Mit16- D9Mit55/ D9Mit125- D9Mit281 Based on genetic mapping, we constructed for a YAC contig across cir region. They covered the entire region or cir and cir gene was located on between the lactotransferrin (ltf) and the macrotubule-associated protein (map4). It is known that sr gene is localized in 64cM of mouse chromosome 9. The two mouse were found to be allelic by complementation test. Recently the spinner mouse has been mapped to our cir region, and tmie gene were elucidated. And further study will be needed in circling mouse to prove tmie gene mutaiton.

• Journal of Microbiology and Biotechnology
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• v.17 no.4
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• pp.638-643
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• 2007

The full encoding sequence for human type II hexokinase (HXK II) was cloned into the E. coli expression vector pET 21b and expressed as a C-terminally hexahistidine-tagged protein in the BL2l (DE3) strain. The IPTG-induced HXK II approximately accounted for 17% of the total E. coli proteins, and 81% of HXK $II_{6{\times}His}$ existed in inclusion bodies. To improve the production of soluble recombinant HXK II protein, in the functionally active form, we used low temperature, and the osmotic stress expression method. When expressed at $18^{\circ}C$, about 83% of HXK $II_{6{\times}His}$ existed in the soluble fraction, which amounted to a 4.1-fold yield over that expressed at $37^{\circ}C$. The soluble form of HXK $II_{6{\times}His}$ was also highly produced in the presence of 1M sorbitol under the standard condition $(37^{\circ}C)$, which indicated that temperature downshift and low water potentials were required to improve the yield of active recombinant HXK II protein. The expressed protein was purified by metal chelate affinity chromatography performed in an IDA Excellose column charged with $Ni^{2+}$ ions, resulting in about 40mg recombinant HXK II protein obtained with purity over 89% from 51 of E. coli culture. The identity of HXK $II_{6{\times}His}$ was confirmed by Western blotting analysis. Taken together, using the stress-governed expression described in this study, human active HXK II can be purified in sufficient amounts for biochemical and biomedical studies.

• Journal of Animal Science and Technology
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• v.56 no.5
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• pp.18.1-18.5
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• 2014

Background: A six-week study involving two hundred and fifty (250) Harco Black layer birds at point of lay was conducted to investigate the effects of potential natural colorant on performance and Egg quality traits. The birds were assigned to five (5) dietary treatments, each containing supplements either of control, Baobab Leaf (BL), Waterleaf (WL), Red Pepper (RP), Canthaxanthin (CTX) at 40 g/kg feed and 50 mg/kg feed of natural and commercial colorants, respectively. Results: Performance records shows that there was no significant (p > 0.05) difference in feed intake across the supplements of Red pepper, Water leaf, Canthaxanthin and control diet, however, birds fed Baobab leaf treatment had a significantly lower (p < 0.05) feed intake value (94.07 g) when compared with other treatments. Body weight gain and Hen Day Production were not significant influenced (p > 0.05) by the dietary treatments, although laying hens fed Baobab leaf supplement had lowest mean HDP of 48.80%, while birds fed Red pepper and Water leaf supplement had an average value of 52.79%. There was no significant effect (p > 0.05) of colorants on egg external traits, compared with the control; birds fed Canthaxanthin treatment had higher mean egg weight (51.79 g), egg length (4.55 g), egg breadth (3.29 g); Red pepper treatment had highest mean shell thickness (0.29 g), however these differences were not significant (p > 0.05). Yolk height, Albumen height, Yolk index, and Haugh unit were not significantly affected (p > 0.05) across treatments. Yolk width was lowest (p < 0.05) in Baobab leaf treatment (2.54 cm); Red pepper, Water leaf and Canthaxanthin (2.89 cm, 2.62 cm and 2.89 cm respectively) were not significantly (p > 0.05) different from the control (2.73 cm). Yolk colour score was significantly highest (p < 0.05) in Red pepper treatment (7.50); Water leaf, Baobab leaf and Canthaxanthin ranged between 2.25-3.31 on the DSM yolk colour fan, Control treatment had the lowest yolk colour score (p < 0.05) of 1.31. Conclusion: The study showed Red pepper as a worthy alternative to commercial yolk colorant. Water leaf and baobab are not good substitutes for canthaxanthin as a yolk colourant.

• The Journal of Korean Medicine
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• v.30 no.1
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• pp.51-63
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• 2009

Objectives: Peroxynitrite ($ONOO^-$), superoxide anion radical (${\cdot}{O_2}^-$ and nitric oxide (NO) are cytotoxic because they can oxidize several cellular components such as proteins, lipids and DNA. They have been implicated in the aging processes, and age-related diseases such as Alzheimer's disease, rheumatoid arthritis, cancer, diabetes, obesity and atherosclerosis. The aim of this study was to investigate the $ONOO^-$, NO, ${\cdot}{O_2}^-$ scavenging and NF-${\kappa}B$ related anti-inflammatory activities of Sotosaja-hwan in ob/ob mice. Methods: Mice were grouped and treated for 5 weeks as follows. Both the normal lean (C57/BL6J black mice) and control obese (ob/ob mice) groups have received standard chow. The experimental groups were fed with a diet of chow supplemented with 30 and 90 mg Sotosaja-hwan per 1 kg of body weight for 14 days. For this study, the fluorescent probes, namely 2',7'-dichlorodihydrofluorescein diacetate (DCFDA), 4,5-diaminofluorescein (DAF-2) and dihydrorhodamine 123 (DHR 123) were used. Western blotting was performed using anti-phospho-$I{\kappa}B$-${\alpha}$, anti-IKK-${\alpha}$, anti-NF-${\kappa}B$ (p50, p65), anti-COX-2, anti-iNOS, anti-YCAM-1 and anti-MMP-9 antibodies, respectively. Results: Sotosaja-hwan inhibited the generation of $ONOO^-$, NO and ${\cdot}{O_2}^-$ in the lipopolysaccharide (LPS)-treated mouse kidney postmitochondrial fraction in vitro. The generation of $ONOO^-$, NO, ${\cdot}{O_2}^-$ and PGE2 were inhibited in the Sotosaja-hwan-administered ob/ob mice groups. The GSH/GSSG ratio was decreased in the ob/ob mice, whereas the ratio was improved in the Sotosaja-hwan-administered groups. Sotosaja-hwan inhibited the protein expression levels of phospho-$I{\kappa}B$-${\alpha}$, IKK-${\alpha}$, NF-${\kappa}B$ (p50, p65), COX-2, iNOS, YCAM-1 and MMP-9 genes. Conclusions: These results suggest that Sotosaja-hwan is an effective $ONOO^-$, ${\cdot}{O_2}^-$ and NO scavenger and has NF-kB related anti-inflammatory activity in ob/ob mice. Therefore, Sotosaja-hwan might be a potential therapeutic drug against the inflammation process and inflammation-related diseases.

• Microbiology and Biotechnology Letters
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• v.30 no.1
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• pp.51-56
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• 2002

An antitumor substance was purified from the culture filtrate of phytopathogenic fungus Alternaria brassicicola SW-3 isolated from soil of a chinese cabbage patch, and its characteristics were investigated. Antitumor activity of A. brassicicola SW-3 was measured by MTT assay. The cytotoxic activity against human cancer cell line was detected in the culture filtrate of A. brassicicola SW-3, but no activity found in mycelium. Antitumor substance was isolated from the culture broth by ethyl acetate extraction and purified by silica gel column chromatography. Structure of the purified compound was analyzed by the instrumental analysis such as $^1$H-NMR, $^{13}$ C-NMR and IR spectroscopy. The purified fungal metabolite of an A. brassicicola SW-3, consists of 11 carbon chain with two hydroxyl groups and two epoxides which is identical to depudecin. The $IC_{50}$/ values of the active compound identified as depudecin were $69\mu$g/mL and $57\mu$g/mL against mouse melanoma B16BL6 cell line, and human hepatoma SK-HEP1 cell line, respectively.

• Proceedings of the Ginseng society Conference
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• 2002.10a
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• pp.129-144
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• 2002

We investigated anti-hyperglycemic and anti-obese effects of Panax ginseng berry extract and its major constituent, ginsenoside Re, in obese diabetic C57BL/6J ob/ob mice and their lean littermates. Animals received daily intraperitoneal injections of Panax ginseng berry extract for 12 days. On Day 5, 150 mg/kg extract-treated ob/ob mice had significantly lower fasting blood glucose levels compared to vehicle-treated mice $(156{\pm}9.0\;mg/dl\;vs.\;243{\pm}15.8mg/dl,$ P<0.01). On Day 12, the extract-treated ob/ob mice became normoglycemic $(137{\pm}6.7\;mg/dl)$ and had significantly improved glucose tolerance. The overall glucose excursion during the two-hour intraperitoneal glucose tolerance test (IPGTT), calculated as area under the curve (AUC), decreased by $46\%$ (P<0.01) compared to vehicle-treated ob/ob mice. Glucose levels of lean mice were not significantly affected by the extract. The improvement in blood glucose levels in 150 mg/kg extracttreated ob/ob mice was associated with significant reduction in serum insulin levels of fed and fasting mice. Consistent with an improvement in insulin sensitivity, hyperinsulinemic euglycemic clamp study revealed a more than 2-fold increase in the rate of insulin-stimulated glucose disposal in treated ob/ob mice $(112{\pm}19.1\;vs.\;52{\pm}11.8{\mu}mol/kg/min$ for the vehicle group, P<0.01). In addition, 150 mg/kg extract-treated ob/ob mice, but not the lean mice, lost significant weight (from $51.7{\pm}1.9g\;on\;Day\;0\;to\;45.7{\pm}1.2$ on Day 12, P<0.01 compared to vehicle-treated ob/ob mice), associated with a significant reduction in food intake (P<0.05) and a very significant increase in energy expenditure (P<0.01) and body temperature (P<0.01). A 12-day treatment with 150 mg/kg Panax ginseng berry extract also significantly reduced plasma cholesterol levels in ob/ob mice. Additional studies demonstrated that ginsenoside Re, a major constituent of the ginseng berry, but not from the root, plays a significant role in anti-hyperglycemic action. This anti-diabetic effect of ginsenoside Re was not associated with body weight changes, suggesting that other constituents in the extract have distinct pharmacological mechanisms on energy metabolism. The identification of a significant anti-hyperglycemic activity in ginsenoside Re may provide an opportunity to develop a novel class of anti-diabetic agent.

• Journal of the East Asian Society of Dietary Life
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• v.13 no.2
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• pp.98-110
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• 2003

This study investigated the nutritional status and the knowledge of the 204 elderly in Young-nam area. Their weight and height were measured and their dietary intake and nutritional knowledge evaluated. The sutjects were 74.7$\pm$5.9 years old in average. The BMIs were in the moderate range. Weights and BMI decreased significantly with age. The dietary assessment data showed that each energy intake of the males and the females was 90.9% and 97.0% compared with the Korean RDA, respectively. The dietary intake of vitamin A, calcium and iron was lower than that of the Korean RDA. The fiber intake of the subjects was 7.4g~7.8g. The MAR of vitamin $_{B2}$ was the lowest, 0.66 and that of phosphorus was the highest, 0.99. According to the nutritional knowledge level, the intake of protein, vitamin A, vitamin $B_1$and iron in the excellent group was significantly higher than that in the poor group. The correlation analysis revealed that the score of nutritional knowledge was positively associated with BMI, MAR and protein intake, while negatively with age.e.

• Radiation Oncology Journal
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• v.24 no.1
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• pp.51-57
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• 2006

Puroose: We examined whether intratumoral (i.t.) administration of dendritic cells (DCs) into a treated tumor could induce local and systemic antitumor effects in a mouse tumor model. Methods and Materials: C57BL/6 mice were inoculated s.c. in the right and left thighs with MCA-102 fibrosarcoma cells on day 0 and on day 7, respectively. On day 7, the tumors (usually 6 mm in diameter) on the right thigh were heated by immersing the tumor-bearing leg in a circulating water bath at $43^{\circ}C$ for 30 min; thereafter, the immature DCs were i.t administered to the right thigh tumors. This immunization procedure was repeated on days 7, 14 and 21. The tumors in both the right and left thighs were measured every 7 days and the average sizes were determined by applying the following formula, tumor $size=0.5{\times}(length+width)$. Cytotoxicity assay was done to determine tumor-specific cytotoxic T-lymphocyte activity. Results: Hyperthermia induced apoptosis and heat shock proteins (HSPs) in tumor occurred maximally after 6 hr. For the local treated tumor, hyperthermia (HT) alone inhibited tumor growth compared with the untreated tumors (p<0.05), and furthermore, the i.t. administered DCs combined with hyperthermia (HT + DCs) additively inhibited tumor growth compared with HT alone (p<0.05). On the distant untreated tumor, HT alone significantly inhibited tumor growth (p<0.05), and also HT + DCs potently inhibited tumor growth (p<0.001); however, compared with HT alone, the difference was not statistically significant. In addition, HT + DCs induced strong cytotoxicity of the splenocytes against tumor cells compared to DCs or HT alone. Conclusion: HT + DCs induced apoptosis and increased the expression of HSPs, and so this induced a potent local and systemic antitumor response in tumor-bearing mice. This regimen may be beneficial for the treatment of human cancers.