• Asian-Australasian Journal of Animal Sciences
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• 제17권5호
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• pp.617-620
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• 2004

The present study compares development of rabbit embryos generated using different oocyte activation protocols and reconstructed with embryonic or cumulus cells as nuclear donor. In vivo matured oocytes were collected from New Zealand White rabbits at 16 h after ovulation treatment and were activated at18 h of post-ovulation treatment. The following schemes of oocytes activation were tested: 1) single electric pulse (EP, 3.2 kV/cm, 3${\times}$20 $\mu$s, 0.3 M mannitol)+5 min culture in the presence of 5 mM Ionomycin, 2) single electric pulse (EP, 3.2 kV/cm, (${\times}$20 $\mu$s, 0.3 M mannitol)+1 h culture in the presence of 2 mM 6-DMAP, and 3) three electric pulses 30 min apart. Cleavage rate, percentage of expanded and hatched blastocysts as well as total cell number of blastomeres of parthenogenetic embryos were significantly higher using either EP+6-DMAP or 3${\times}$EP schemes, comparing with EP+Ionomycin. Development rate up to hatched blastocyst stage of cloned rabbit embryos using the EP+6-DMAP for activation of nuclei were 19% for embryonic cell nuclei and 36% for cumulus cell nuclei. The best activation protocol optimalized in this study was the combined treatment "P+6-DMAP" which may be potentially used for nuclear transfer protocol.

• 한국시뮬레이션학회:학술대회논문집
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• 한국시뮬레이션학회 1999년도 춘계학술대회 논문집
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• pp.6-10
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• 1999

We present a simulation result to the analysis of the effects of cell residence time distributions upon the expected channel occupancy time based on an analytic mobility model. Numerical examples show that exponential distribution provides upper and lower bound to the expected channel occupancy times of new calls and handoff calls. This fact reveals that the assumption of exponential distribution as the cell residence time distribution as the cell residence time distribution may over- or under-estimate cellular mobile systems.

• 한국가축번식학회지
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• 제11권3호
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• pp.161-169
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• 1987

This experiment was carried out to investigate the optimal cooling rate and the plunging temperature into liquid nitrogen of the 8-cell hamster embryos. The female hamsters were induced to superovulate by intraperitoneal injection of 30 i.u. PMSG. Embryos were flushed from oviduct and uterine horn. Embryos were frozen and incubated with a modified Dulbecco's phosphate buffered saline, and equilibrated with 1.5M-dimethyl sulfoxide by a 3-step procedure. The cooling rate of samples was 1$^{\circ}C$/min from room temperature to -5$^{\circ}C$ and the samples were seeded at -5$^{\circ}C$. The plunging temperatures into liquid nitrogen were -20, -25, -30, -35, -40, -45, -50 and -55$^{\circ}C$ at 0.3$^{\circ}C$/min, 0.5$^{\circ}C$/min and 1$^{\circ}C$/min cooling rates, respectively. This mean numbers of ovulation points and recovered embryos were 59.4 and 48.4 appearing 81.6% recovery rate. The percentage of 8-cell embryos in recovered embryos was 68.2. The survival rates of embryos plunged at -45$^{\circ}C$ were 73.5% at 0.3$^{\circ}C$/min, 44.8% at 0.5$^{\circ}C$/min and 30.3% at 1$^{\circ}C$/min cooling rates, respectively. This mean numbers of ovulation points and recovered embryos were 59.4 and 48.4 appearing 81.6% recovery rate. The percentage of 8-cell embryos in recovered embryos was 68.2. The survival rates of embryos plunged at -45$^{\circ}C$ were 73.5% at 0.3$^{\circ}C$/min, 44.8% at 0.5$^{\circ}C$/min and 30.3% at 1$^{\circ}C$/min cooling rates, respectively. The survival rates at 0.3$^{\circ}C$/min were significantly high. Under the condition of 0.3$^{\circ}C$/min cooling rate, the survival rates of embryos according to the plunging temperature were 70.0% and 73.5% at -40 and -45$^{\circ}C$, and those were higher than other plunging temperatures. Under the condition of 0.5$^{\circ}C$/min and 1$^{\circ}C$/min cooling rates, the survival rates according to the plunging temperatures were lower than the cooling rate of 0.3$^{\circ}C$/min, showing the similar tendency at all the plunging temperatures. In conclusion, 8-cell hamster embryos showed the best survival rates at 0.3$^{\circ}C$/min cooling rate and -45$^{\circ}C$ plunging temperature.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2002년도 생물공학의 동향 (X)
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• pp.293-296
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• 2002

Hyperosmotic pressure increased specific antibody productivity ($q_{Ab}$) of recombinant CHO cells (SH2-0.32) while it depressed cell growth. Thus, the use of hyperosmolar medium did not increase the maximum antibody concentration substantially. To overcome this drawback, the feasibility of biphasic culture strategy was investigated. In the biphasic culture, cells were first cultivated in the standard medium with physiological osmolality(294 mOsm/kg) for cell growth. When cells reached the late exponential phase of growth, the spent standard medium was replaced with the fresh hyperosmolar medium (522 mOsm/kg) for antibody production. The ($q_{Ab}$) in growth phase with the standard medium was 2.1 ${\mu}g/10^6cell/day$ while the ($q_{Ab}$) in antibody production phase with the hyperosmolar medium (522 mOsm/kg) was 11.1 ${\mu}g/10^6cell/day$. Northern blot analysis showed a positive relationship between the relative contenet of Ig mRNA and ($q_{Ab}$), indicating that transcriptional regulation was involved in the response of rCHO cells to hyperosmotic pressure. Due to the enhanced ($q_{Ab}$) and increased cell concentration in biphasic culture, the maximum antibody concentration obtained in biphasic culture with 522 mOsm/kg medium exchange was 161% higher than that obtained in batch culture with the standard medium. Taken together, simple biphasic culture strategy based on hyperosmotic culture for improved foreign protein production from rCHO cells is effective in improving antibody production of rCHO cells.

• Journal of Microbiology and Biotechnology
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• 제17권6호
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• pp.1036-1040
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• 2007

Sodium butyrate (NaBu) has been used to enhance protein expression levels in mammalian cell culture. To determine the clonal variability of recombinant Chinese hamster ovary (rCHO) cells in response to NaBu addition regarding specific antibody productivity $(q_{Ab})$, three rCHO clones were subjected to different concentrations of NaBu. For all three clones, NaBu addition inhibited cell growth and decreased cell viability in a dose-dependent manner. On the other hand, the enhancing effect of NaBu on $q_{Ab}$ varied significantly among the clones. NaBu addition enhanced the antibody production of only one clone. RT-PCR analysis revealed that the changes in $q_{Ab}$ correlated linearly with those of the mRNA transcription level. Thus, it was concluded that the different enhancing effects of NaBu on protein expression in rCHO cell clones resulted from their different mRNA transcription levels.

• Clinical and Experimental Reproductive Medicine
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• 제25권2호
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• pp.207-216
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• 1998

Certain types of somatic cells, as well as follicular cumulus cells associating with mammalian oocytes, are known to produce beneficial effects on in vitro fertilization and pre implantation development of mammalian eggs when they are present in oocyte culture medium. To investigate the nature of the effects of somatic cells, the resistance of mouse oocytes against chymotrypsin treatment was examined after culture within various cell conditioned media. When mouse oocytes matured for 17-18 hr in the presence of cumulus cells were treated with 1 % chymotrypsin, half of them remained still alive even after 240 min $(t_{50}>240.0)$. In contrast half of mouse oocytes cultured without cumulus cells underwent degeneration within 65.0 min $(t_{50}=65.0{\pm}13.2min)$ of the same treatment. To see if the effects were duc to the secretory products of cumulus cells, mouse cumulus cells were cultured for 20 hr in medium containing 0.4% BSA and the supernatant of culture medium (conditioned medium) was taken. After maturation in the cumulus cell conditioned medium, mouse oocytes exhibited $t_{50}=190.0{\pm}10.8$ min upon chymotrypsin treatment whereas half of oocytes cultured without conditioned medium degenerated within 25.5 min. Human granulosa cell conditioned medium gave similar effects such that oocytes matured in conditioned medium exhibited $t_{50}=183.3{\pm}19.1$ min while $t_50$ of control group oocytes was $60.0{\pm}6.8$ min, Oocytes matured in vero cell conditioned medium exhibited $t_{50}=196.7{\pm}8.8$ min. On the other hand, amniotic cell conditioned medium resulted in the chymotrypsin resistance of $t_{50}=80.0{\pm}8.4$ min which was not statistically different from the control value of $t_{50}=48.0{\pm}13.2$ min. Based upon these results, it is suggested that certain somatic cell types including cumulus cells might change the biochemical properties of mouse oocyte membrane during meiotic maturation as revealed by the enhanced resistance against chymotrypsin treatment. Such effects of somatic cells appear to be mediated via the secretory products rather than direct communication between somatic cells and oocytes.

• 한국축산식품학회지
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• 제42권1호
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• pp.142-152
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• 2022

The present study was aimed at examining the antibacterial effects of nonthermal decontamination processes, which are equivalent to thermal treatment, to ensure microbiological safety of raw ground chicken. Escherichia coli or Salmonella were inoculated into 25 g of raw ground chicken samples. The raw ground chicken samples were non-treated or treated with high hydrostatic pressure (HHP) at 500 MPa (1-7 min), light-emitting diode (LED) irradiation at 405 nm wavelength (30-120 min), and heat at 70℃, 90℃ (1-60 min), and 121℃ (1-15 min). E. coli and Salmonella cell counts were enumerated after treatments. Moreover, the color parameters of treated raw ground chicken were analyzed. HHP treatment reduced E. coli and Salmonella cell counts by more than 5 Log CFU/g and more than 6 Log CFU/g after 7 min and 1 min, respectively; these effects were equivalent to those of thermal treatment. However, LED irradiation reduced Salmonella cell counts by only 0.9 Log CFU/g after 90 min of treatment, and it did not reduce E. coli cell counts for 90 min. Compared with those of the non-treated samples, the ΔE (total color difference) values of the samples treated with HHP were high, whereas the ΔE values of the samples treated with LED irradiation were low (1.93-2.98). These results indicate that despite color change by HHP treatment, HHP treatment at 500 MPa could be used as a non-thermal decontamination process equivalent to thermal treatment.

• 공업화학
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• 제21권3호
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• pp.301-305
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• 2010

본 연구에서는 도금전처리 공정에서 사용되는 다양한 조성의 맞춤형 알칼리계 탈지제를 개발하고, 탈지제의 수명을 예측함으로 효율을 높임과 동시에 공정품의 불량을 줄이고자 하였다. 알칼리계 탈지제는 침지형 탈지제로 NaOH, $Na_2CO_3$, $Na_2SiO_3$, SLS 등을 일정비율로 첨가하여 제조하였으며, 제조된 탈지제의 성능평가를 위해 알칼리계 탈지액의 온도를 $40{\sim}50^{\circ}C$로 설정하여 30~40 min 동안 오염된 철 시편을 넣은 후 오염물을 제거하였다. 오염물이 제거된 시편은 물방울 맺힘실험과 Hull-cell 도금분석을 통해 탈지제의 성능 및 수명을 확인하였다. 최적 탈지제의 조성은 NaOH (30 g/L) + SLS (6.0 g/L) + $Na_2SiO_3$ (2.0 g/L) + $Na_2CO_3$ (40 g/L)을 추천할 수 있으며, 물방울맺힘 실험과 Hull-cell 도금 분석 결과 최적 탈지조건은 $50^{\circ}C$, 35 min인 것으로 나타났다.

• 대한본초학회지
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• 제28권6호
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• pp.155-160
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• 2013

Objectives : Gastric cancer is a leading cause of cancer-related deaths, worldwide. Houttuynia cordata Thunberg (H. cordata) has been used as a medicinal plants and it has an anti-cancer activity in human colorectal cancer and leukemic cancer. However, the potential anti-cancer activity and mechanisms of H. cordata for human gastric cancer cells have not been tested so far. Thus, this study examined the biological effects of H. cordata on the human gastric cancer cell line SNU-1 and AGS. Methods : Inhibition of cell proliferation and cell cycle by H. cordata was carried out by MTT assay and Muse cell cycle analysis and the expressions of protein associated with apoptosis and cell cycle regulation were investigated with Western blot analysis. Results : In MTT assay, the proliferation of SNU-1 and AGS cells was significantly inhibited by H. cordata in a time and dose dependent manner, Inhibition of cell proliferation by H. cordata was in part associated with apoptotic cell death, as shown by changes in the expression ratio of Bax to Bcl-2 by H. cordata. Also, H. cordata regulated the expression of cell cycle regulatory proteins such as pRb, cyclin D1, cyclin E, CDK4, CDK2, p21 and p15. Conclusion : The antiproliferative effect of H. cordata on SNU-1 and AGS gastric cancer cells revealed in this study suggests that H. cordata has intriguing potential as a chemopreventive or chemotherapeutic agent.

• Radiation Oncology Journal
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• 제21권4호
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• pp.306-314
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• 2003

목적: K562 세포의 방사선에 의한 세포 사망은 mitotic catastrophe 현상이 위주로 나타나지만 herbimycin A (HMA)에 의하여 apoptosis 반응이 촉진되는 반면 genisteln에 의하여 두 가지 형태의 세포사망이 모두 억제된다. 본 연구에서는 HMA와 genistein에 의한 K562세포의 방사선 유도 세포주기 조절 변화와 세포 사망 양상의 연관성을 조사하였다. 대상 및 방법: 지수증식기의 KS62 세포에 6 MV 선형가속기(Clinac 1,m C, Varian)를 이용하여 200~300 cGy/min의 선량률로 10 Gy를 균일하게 조사하였다. HMA와 genistein은 각각 250 nM와 25$\mu$M농도로 방사선 조사 후 즉시 투여하였다. 실험에서는 세포주기, 오절인자의 발현 및 활성, 노화 및 분화정도 등에 있어서의 시간에 따른 변화를 조사하였다. 결과: 방사선 단독조사에서 KS62세포는 G2기의 정체를 보였으나 정상적인 053을 가지는 세포와는 달리 지속적인 세포주기의 정체를 보이지 않았다. G2정체가 유지되는 동안 cyclin Bl의 점진적인 증가를 관찰할 수 있었으며, 이는 염색체의 복제가 완료되지 않은 상태에서 M기로 진행하여 미성숙한 염색체 응축과 mitotic catastrophe 현상이 나타나는 것과 일치한다. 방사선 조사와 함께 HMA를 투여한 경우에는 G2정체가 빠르게 해소되었으며 동시에 Gl기에서 세포가 정체되는 양상을 보였다. 세포주기 조절인자 cdc2 kinase 활성 증가와 cyclln I와 A 발현 및 CDK2 활성의 감소 등의 현상으로 설명되며, 이는 apoptosis의 증가와 연관성을 갖는다. 반면 genistein의 경우에는 cyclin Bl과 떨cfsc 발현 및 cdc2활성이 모두 감소하는 등 G2정체를 계속 유지하였다. 이와 함께 방사선에 의한 노화와 megakaryocyte로의 분화도 지속되는 것을 관찰할 수 있었다. 결론: HMA와 genistein에 의한 KS62세포의 방사선 유도 세포사망의 변화는 세포주기 조절과 밀접하게 연관되어 있음을 확인하였다. 이는 다양한 방사선 유도 세포사망의 기전을 이해하는 데 독창적인 모델을 제공하며, 방사선을 이용한 암 치료법의 개발에 새로운 표적을 제공할 수 있을 것이다.