• Journal of Embryo Transfer
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• v.20 no.2
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• pp.89-95
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• 2005

In this study, we investigated the effects of number, grade, stage and day of embryos on pregnancy and abortion rates of Holstein heifer recipients after transfer of in-vitro produced Korean Native Cattle (Hanwoo) embryos. When one, two or three blastocysts were transferred to recipients, pregnancy (32.0, 44.0 and $32.0\%$) and abortion (12.5, 13.6 and $6.3\%$) rates did not differ among treatments. The pregnancy and abortion rates of different grades of blastocysts were $40.8\~44.2\%\;and\;9.4\~16.3\%$, respectively. However, they were not significantly different. According to the stage of blastocyst, the pregnancy rates were similar between $39.1\%\;and\;45.3\%$. However, the abortion rate in hatched blastocyst stage $(20.8\%)$ was significantly higher than that in late blastocyst stage $(9.0\%)$. The pregnancy rates in day $7\cdot7$ and day $7\cdot8$ blastocyst (47.1 and $53.9\%$, respectively) was significantly higher than that in the day $9\cdot9\;and\;8\cdot9$ blastocyst (29.8 and $27.7\%$, respectively). The abortion rate was the highest in the day $9\cdot9$ blastocyst $(30.8\%)$ among treatments, and it was significantly different compared to that in the day $7\cdot7$ blastocyst $(10.1\%)$. Therefore, this study suggested that the pregnancy rate was affected by only day of embryos, while the abortion rate was affected by stage and day of embryos.

• Food Science of Animal Resources
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• v.20 no.2
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• pp.101-106
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• 2000

Effects of Ph on the activity of lipase isolated from milk fat globules were investigated, using coconut oil and homogenized milk as substrate. With buttermilk as an enzyme source for coconut oil and homogenized milk substrates bell-shaped curve was observed at $37^{\circ}C$, having the highest activity at pH 9.5. However, lipase activity at $0^{\circ}C$ continuously increased up to pH 10.0. With the purified lipase for homogenized milk substrate, the bell -shaped curve and the highest activity were observed at $37^{\circ}C$ and pH 9.0, respectively. Lipase activity at $0^{\circ}C$ increased up to pH 10.0. The addition of bovine serum albumin to the coconut oil shifted the optimum pH to pH 9.5 and the activity remarkably declined at pH 10.0. The effect of pH on the stability of purified lipase was depending on the temperature. Wehn the lipase kept at $37^{\circ}C$ for 20 minutes, it's activity remarkably declined as pH increased: the activity at pH 10.0 was declined by 13% of that pH 8.5. However, when the lipase kept at $4^{\circ}C$ for 60minutes, the activity was stable within the range of pH 7.5 to 10.0.

• Food Science of Animal Resources
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• v.23 no.2
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• pp.137-144
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• 2003

This study was carried out to investigate safety evaluation of IGEs separated and refined from bovine milk and commercial recombinant human IGFs. In order to evaluate toxicity of these samples, acute toxicity test and short term toxicity test were investigated with IGF-I separated and refined from colostrum and commercial recombinant human IGF-I from R&D systems company. for acute toxicity test, we selected recombinant human IGF-I from R&D systems company and establish one control group and three dose-level groups(0, 10, 20 and 50 $\mu\textrm{g}$ per rat). We have intravenously injected tail of rats with selected sample once. After 20 days, pathological cellular tissue analyses were investigated with liver, kidney and spleen of 12 rats in all test groups. However, Morbid tissue and abnormal statistical results were not discovered in all cellular tissues. For short term toxicity test, we selected IGF-I separated and refined from colostrum and establish one control group and three dose-level groups(0, 5, 10 and 15 $\mu\textrm{g}$/day per rat). Rats were orally injected with selected sample once a day during two weeks. After short term toxicity test period, Pathological cellular tissue analyses were investigate with liver, kidney and spleen of 12 rats in all test groups. However, Morbid tissue and abnormal statistical results were not discovered in all cellular tissues. These results suggest that IGF-I treated groups show no significant toxicological findings with changes of body weight, food consumption, water consumption, and pathological findings compared with control groups.

• Food Science of Animal Resources
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• v.23 no.4
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• pp.350-355
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• 2003

Raw skim milk and colloidal calcium phosphate-free skim milk were treated with microbial transglutaminase (TGase), ultracentrifuged at varying rates and were observed to contain textural properties using a scanning electron microscope (SEM). Skim milk showed irregular signs of conformation at lower centrifugal rate, and associated regular (10,000 ${\times}$g) and thin with broad holes (20,000 ${\times}$g). The associated texture became thick and irregular (40,000 ${\times}$g), and fine particles were regularly associated (100,000 ${\times}$g). When skim milk was incubated for 1 hr with TGase, casein micelles aggregated and broadened as centrifugation rate increased. When skim milk was incubated for 8 hrs with TGase, casein micelles associated to large widened aggregates, and were associated regularly which then became irregular (100,000 ${\times}$g). When colloidal calcium phosphate-free skim milk incubated for 1 hr with TGase showed no sediment, the milk incubated for 8 hrs with TGase associated together, yielding broadened and regular layers as the centrifugation rate increased. It is assumed that such phenomena could be caused by protein crosslinking reaction with TGase and conformational change of casein molecules, as well as dependencies on reaction time, temperature and ultracentrifugation rate.

• Journal of Embryo Transfer
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• v.24 no.1
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• pp.21-27
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• 2009

This study was performed to elucidate the effects of addition of ${\beta}-lactoglobulin$ and bovine serum albumin (BSA) in vitro maturation (IVM) and in vitro culture (IVC) medium on porcine embryo production. The development rate to the 2 cell ($71.4{\sim}75.6%$) and blastocyst stages ($6.8{\sim}13.3%$) with different BSA concentrations in IVM medium were similar among treatment groups. Blastocyst hatching rate was significantly higher in the control group (0.0mg/ml) than in the group of 1.0mg/ml supplement (20.0% vs. 0.0%; p<0.05). The development rate to the 2 cell ($62.0{\sim}70.6%$) and blastocyst stages ($15.4{\sim}38.5%$) with different ${\beta}-lactoglobulin$ concentrations in IVM medium was similar among treatment groups. The development rate to the blastocyst was significantly higher in the group of 1.0mg/ml(15.3%) than in the group of 0.5mg/ml supplement (7.6%, p<0.05). The development rate to the 2 cell and blastocyst stages following the first addition of ${\beta}-lactoglobulin$ in IVM medium was significantly higher in the control group (77.0% and 18.9%) and was $0{\sim}44\;hr$(77.2% and 16.9%) greater than that observed in other treatment groups (p<0.05). The development rate to the 2 cell stage ($68.1{\sim}74.8%$) and blastocyst stages ($9.2{\sim}12.7%$) with different BSA concentrations in IVC medium was similar among treatment groups. However, blastocyst hatching rate was significantly higher in the group of 3.0mg/ml supplement (30.0%) than in the control group (0.0%; p<0.05). The development rate to the 2 cell stage ($72.9{\sim}78.0%$), blastocyst ($7.1{\sim}14.2%$) and hatching stages ($33.3{\sim}38.1%$) were not different. The development rate to the 2 cell stage ($63.6{\sim}72.5%$), blastocyst ($8.4{\sim}16.1%$) and hatching stages ($18.2{\sim}37.5%$) at the different culture periods were similar among treatment groups. This study suggested that if the addition level and periods of ${\beta}-lactoglobulin$ addition are adjusted, it is possible to replace BSA in the in vitro porcine embryo production.

• Journal of Life Science
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• v.20 no.6
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• pp.914-921
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• 2010

In this study, we carried out a series of experiments to know whether melatonin, an anti-oxidative and immunosuppressive agent, played an important role in endothelial cells. It was revealed that melatonin had little or no effect on endothelial proliferation, cell death or migration. Additionally, melatonin had no effect on adhesion of THP-1 leukocytes to bovine aortic endothelial cells (BAECs) and THP-1 homotypic cell aggregation. In contrast, it was shown that melatonin diminished the basal level of nitric oxide by PP2A-mediated dephosphorylation of endothelial nitric oxide synthase (eNOS), leading to enhanced detachment of BAEC from the extracellular matrix. Collectively, melatonin in high doses decreases the NO production via regulations of PP2A and eNOS activities, inducing detachment of endothelial cells, a possible initial step for thrombosis.

• Journal of Life Science
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• v.27 no.5
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• pp.553-560
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• 2017

Fermented garlic extract (FGE) is a well-known functional food containing cycloalliin. Here we tested if FGE has an effect on the vascular system. We found that FGE had an effect on monocytic adhesion to endothelial cells, which occurs at the initial step of processes of inflammation. From dose-dependent experiments, 0.1 to $1{\mu}g/ml$ of FGE appeared to inhibit lipopolysaccharide (LPS)-enhanced heterotypic cell adhesion between THP-1 and BAECs. This result indicates that FGE blocks vascular inflammation. Then we found that FGE activates eNOS and Akt in BAECs. The phosphorylation of eNOS was maximally elevated 10 min after FGE treatment. Parallely, the phosphorylation of Akt was also maximally increased 10 min after FGE treatment. Consistently, it was found that FGE enhanced the production of nitric oxide. We then examined whether NO mediates THP-1 cell adhesion to BAECs. Both Akt and eNOS inhibitors appeared to reverse an inhibitory effect of FGE. These findings indicate that FGE inhibits LPS-enhanced heterotypic cell adhesion via Akt and eNOS. In conclusion, FGE plays an important role in prevention of inflammatory diseases.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.26 no.1
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• pp.24-39
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• 2000

Various methods and graft materials have been used to fill in the defect adjacent to the implants and considered as clinically acceptable. But it is not clear whether the regenerated bone increases the implant-bone contact and supports the implant. The purpose of this study is to evaluate regenerated bone surrounding implants using bone morphogenetic protein(BMP) and demineralized freeze-dried bone(DFDB), and the interfaces between implants and regenerated bone. bBMP was extracted and partially purified from the bovine bone matrix using heparine chromatography. Demineralized freeze-dried bone was made from the dog. Inactive insoluble collagenous bone matrix(IBM) of dog was used as carrier of bBMP. Interfaces of titanium coated epoxy resin implants were processed for demineralized section for transmission electron microscopy(TEM) and those of screw type implants were for nondemineralized section for light and fluoromicroscopic examination. Implants were inserted in the inferior border of mandible of adult dogs and artificial bony defects($3{\times}3{\times}4mm$) were made at the mesial and distal side of implants. Defects were filled with BMP(BMP group) and DFDB(DFDB group). For the fluoromicroscopic examination, the fluorescent dyes(oxytetracycline, calcein green, alizarin red) were injected 2, 4, 6, 8, 12 weeks after implantation. The experimental animals were sacrificed at the 6th and the 12th week and their mandible were extirpated and processed for examination with light microscopy, fluoromicroscopy and TEM. The obtained results were as follows : 1. By the light microscopic findings, the defects were filled with woven bone at the 6th week and compact bone at the 12th week, and the osseointegrations were seen in both groups. There was no histological difference between them. 2. On the basis of the histomorphometric analysis, BMP group(6th week: 40.25%, 12th week: 56.04%) had higher bony contact ratio than DFDB group(38.37%, 42.63%). There was significant difference between two groups at the 12th week(p<0.05). 3. The amount of bone formation in BMP group was more prominent than in DFDB group. Significant difference was noted among two groups at the 6th and the 8th week(p<0.05). 4. By the transmission electron microscopic findings, $0.4-2{\mu}m$ soft tissue layer was found in adjacent to the interfaces and over the collagen fibrils of bone at the 6th week. However, about 100nm amorphous layer was noted at the interface or collagen fibrils directly extended to the titanium surface at the 12th week. There was no significant difference between two groups. 5. These results suggest that BMP and DFDB can be used as good graft materials in the regeneration of bone adjacent to implant, and BMP is more valuable as a bone inducer than DFDB.

• Imaging Science in Dentistry
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• v.38 no.3
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• pp.125-132
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• 2008

Purpose : To investigate the effects of irradiation on transforming growth factor ${\beta}_1$ (TGF-${\beta}_1$) mRNA expression and calcific nodule formation in MC3T3-E1 osteoblastic cell line. Materials and Methods : Cells were cultured in alpha-minimum essential medium ($\alpha$-MEM) supplemented with 10% fetal bovine serum and antibiotics. When the cells reached the level of 70-80% confluence, culture media were changed with $\alpha$-MEM supplemented with 10% FBS, 5 mM $\beta$-glycerol phosphate, and $50\;{\mu}g/mL$ ascorbic acid. Thereafter the cells were irradiated with a single dose of 2, 4, 6, 8 Gy at a dose rate of 1.5 Gy/min. The expression pattern of TGF-${\beta}_1$ mRNA, calcium content and calcific nodule formation were examined on day 3, 7, 14, 21, 28, respectively, after the irradiation. Results : The amount of TGF-${\beta}_1$ mRNA expression decreased significantly on day 7 after irradiation of 4, 6, 8 Gy. It also decreased on day 14 after irradiation of 6, 8 Gy. and decreased on day 21 after irradiation of 8 Gy. The amount of calcium deposition decreased significantly on day 7 after irradiation of 4, 8 Gy (P < 0.01) and showed a decreased tendency on day 14, 21 after irradiation of 4, 6, 8 Gy. The number of calcific nodules was decreased on day 7 after irradiation of 4, 8 Gy. Conclusion: Irradiation with a single dose of 4, 6, 8 Gy influences negatively the bone formation at the molecular level by affecting the TGF-${\beta}_1$ mRNA expression that was associated with proliferation and the production of extracellular matrix in MC3T3-E1 osteoblastic cell line.

• Journal of Periodontal and Implant Science
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• v.28 no.1
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• pp.17-35
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• 1998

Chitosan, with a chemical structure similar to hyaluronic acid, has been implicated as a wound healing agent. The purpose of this research was to evaluate the effects of chitosan on the characteristics of periodontal ligament cells, calvaria cells and gingival fibroblasts and to define the effects of chitosan on bone formation in vitro. In control group, the cells were cultured alone with Dulbecco's Modified Eagle's Medium contained with 10% Fetal bovine serum, 100unit/ml penicillin, $100{\mu}g/ml$ streptomycin, $0.5{\mu}g/ml$ amphotericin-B. In experimental group, chitosan($40{\mu}g/ml$) is added into the above culture condition. And then each group was characterized by examining the cell proliferation at 1,3,5,7,9,12,15 day, the amount of total protein synthesis, alkaline phosphatase activity at 3, 7 day and the ability to produce mineralized nodules of rat calvaria cell at 11 day. The results were as follows : 1. At early time both periodontal ligament cells and calvaria cells in chitosan-treated group proliferated more rapidly than in non-treated control group, but chitosan-treated group of periodontal ligament cells at 9 days and calvaria cells at 12days showed lower growth rate than control group. Gingival fibroblast in chitosan-treated group had lower growth rate than in control group but the difference was not statistically significant (P< 0.01).2. Both periodontal ligament cells and calvaria cells in chitosan-treated group showed much protein synthesis than in control group at 3 days, but showed fewer than in control group at 7 days. Amount of total protein synthesis of gingival fibroblast didn't have statistically significant difference among the two groups(P< 0.01). 3. At 3 and 7 days, alkaline phosphatase activity of periodontal ligament cells and calvaria cells was increased in chitosan-treated group, but at 7 days there was not statistically significant difference among the two groups of calvaria cells (P< 0.01). Alkaline phosphatase activity of gingival fibroblast didn't have statistically significant difference among the two groups(P<0.01). 4. Mineralized nodules in chitosan-treated group of rat calvaria cells were more than in control group. In summery, chitosan had an effect on the proliferation, protein systhesis, alkaline phosphatase activity of periodontal ligament cells and calvaria cells, and facilitated the formation of bone. It is thought that these effects can be used clinically in periodontal regeneration therapy.