• 한국수정란이식학회지
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• 제22권4호
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• pp.265-269
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• 2007

This study was conducted to investigate the efficacy of vitrification procedure for the cryopreservation of porcine oocytes and the utilization of vitrified oocytes as recipient cytoplasts for somatic cell nuclear transfer (NT), and observed that porcine oocytes are evaluated by pronuclear formation, and parthenogenetic development. Single fetal donor cells were deposited into the perivitelline space of vitrified enucleation oocytes, followed by electrical fusion and activation. NT embryos were cultured in NCSU-23 medium supplemented with 5% FBS, at $38.5^{\circ}C$ in 5% $CO_2$ and air. 1. When the developmental rates of the oocytes after being culture for $0{\sim}10$ hours vitrified with EDS and ETS were 42.0%, 38.0%, respectively. This results were lower than the control group(62.2%). 2. When the developmental rates of the oocytes after being culture for $0{\sim}10$ hours vitrified-thawed with sucrose and glucose, 5% PVP, NCSU-23 supplemented with 10% FBS were 33.3%, 25.9%, respectively. This results were lower than the control group(55.6%). 3. The fusion and development to the blastocyst stage between the NT embryos constructed with the vitrified and non-vitrified oocytes were significant differences. Developmental rate of oocytes and NT embryos constructed with the vitrified or non-vitrified oocytes were $13.0{\pm}2.4%\;and\;23.2{\pm}2.4%$, respectively.

• 한국응용곤충학회지
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• 제37권2호
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• pp.187-191
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• 1998

새로운 Bacillus thuringiensis NT0423균주를 이용한 배양체계를 확립하고 대량생산을 위한 새로운 배양배지를 개발하였다 대두박과 밀기울의 다양한 성분비로 구성된 5종의 SW 배지는 기존의 인공합성배지인 GYS나 LB B. huringiensis 의 성장과 포자형성면에서 훨씬 우수한 결과를 보였고, 그중 대두박과 밀기율이 3:2의 비율로 구성된 SW32배지에서 전체 세포수가 3.9$\times$${10}^{8}$CFU/ml에 달했고, 대부분의 세포가 빠르게 포자를 형성하여 (3.7$\times$${10}^{8}$CFU/ml)가장 효율적이었다. 배양에 따른 배지의 pH는 최저 6.2에서 최고 7.3까지 변화하여 성장에 크게 영향을 미치지 않았고, 전체 배양액에서 대두박과 밀기울이 차지하는 비율이 4%일 경우, ml당 4.2$\times$${10}^{8}$CFU/ml의 포자가 형성되어 B. thuringiensis 의 성장에 가장 유리하였다. 교반플라스크와 5ι의 소규모 발효기를 이용한 B. thuringiensis 의 배양조건 확립에 의해 각각 최대 1$\times$${10}^{9}$CFU/ml과 5$\times$${10}^{10}$CFU/ml에 해당하는 많은 양의 균체를 회수할 수 있다.

• Journal of Microbiology and Biotechnology
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• 제17권12호
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• pp.2033-2041
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• 2007

For many years, it has been demonstrated that neurotrophins regulate the adult nervous system, implicating their potential as therapeutic agents for the treatment of neurodegenerative diseases. We generated adenoviral vectors encoding brain-derived neutotrophin factor (BDNF) and neurotrophin-3 (NT3) and tested either separately or together for the ability to induce differentiation of neuronal precursor cells with two different origins. Separate transduction of adenovirus delivering BDNF (BDNF-Ad) or NT3 (NT3-Ad) induced the neuronal differentiation in hippocampal and cortical precursor cells. NT3-Ad infected cells extended short neurites, whereas BDNF-Ad infected cells had longer neurites. In the early differentiation of hippocampal precursor cells, simultaneous infection of BDNF-Ad and NT3-Ad promoted further differentiation and neurite elongation compared with the separate infection of each virus. In contrast, simultaneous infection did not show the synergistic effect in the cortical precursor cells, suggesting that the neurotrophins play distinct roles in different regions of the brain. However, the numbers of neurites and spines per differentiated cells were markedly increased in cortical as well as hippocampal precursor cells, indicating the promotion of efficient neurite elongation and formation of dendritic spine, when BDNF-Ad and NT3-Ad were co-infected. These results suggest more studies in the effect of a combinatorial use of neurotrophins on different sites of brain need to be carried out to develop gene therapy protocols for neurodegenerative diseases.

• Clinical and Experimental Pediatrics
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• 제59권12호
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• pp.477-482
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• 2016

Purpose: We conducted a study to determine which factors may be useful as predictive markers in identifying Kawasaki disease (KD) patients with a high risk of resistance to intravenous immunoglobulin (IVIG) and developing coronary artery lesions (CAL). Methods: We enrolled 287 patients in acute phase of KD at a single center. The demographic, clinical and laboratory data were collected retrospectively. Results: There were 34 patients in the IVIG resistant group. The IVIG resistant group had significantly higher serum N-terminal-pro-brain natriuretic protein (NT-proBNP) levels (P<0.01) and polymorphonuclear neutrophil (PMN) percentage (P<0.01) in comparison to the IVIG responders. The results yielded sensitivity (78.8%, 60.6%), specificity (58.2%, 90%) and cutoff value (628.6 pg/mL, 80.3%) of NT-proBNP and PMN respectively, in predicting IVIG resistance. Despite IVIG administration, 13 of the 287 patients developed CAL. The patients in the CAL group had higher NT-proBNP levels (P<0.01) and higher PMN percentage (P<0.01). In these patients, the results yielded sensitivity (73.3%, 56.7%), specificity (67.9%, 88.9%) and cutoff value (853.4 pg/mL, 80.3%) of NT-proBNP and PMN respectively, for predicting CAL. The area under the curve (AUC) for predicting resistance to IVIG was NT-proBNP 0.712, PMN 0.802. The AUC for predicting CAL was NT-proBNP 0.739, and PMN 0.773. Conclusion: Serum NT-proBNP levels and PMN percentage were significantly elevated in patients with KD with IVIG resistance and CAL. Thus, they may be useful predicting markers for IVIG resistance and development of CAL in KD patients.

• Reproductive and Developmental Biology
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• 제32권3호
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• pp.183-191
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• 2008

This study was conducted to investigate the development and gene expression in miniature pig nuclear transfer (mNT) embryos produced under different osmolarity culture conditions. Control group of mNT embryos was cultured in PZM-3 for 6 days. Treatment group of mNT embryos was cultured in modified PZM-3 with NaCl (mPZM-3, 320 mOsmol) for 2 days, and then cultured in PZM-3 (270 mOsmol) for 4 days. Blastocyst formation rate of the treatment group was significantly higher than the control and the apoptosis rate was significantly lower in treatment group. Bax-$\alpha$ and caspase-3 mRNA expression were significantly higher in the control than the treatment group. Also, the majority of imprinting genes were expressed aberrantly in in vitro produced mNT blastocysts compared to in vivo derived blastocyst H19 and Xist mRNA expression were significantly lower in the control than the treatment group or in vivo. IGF2 mRNA expression was significantly higher in the control than the treatment group or in vivo. IGF2r mRNA expression was significantly lower in the control. Methylation profiles of individual DNA strands in H19 upstream T-DMR sequences showed a similar methylation status between treatment group and in vivo. These results indicate that the modification of osmolarity in culture medium at early culture stage could provide more beneficial culture environments for mNT embryos.

• Reproductive and Developmental Biology
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• 제30권2호
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• pp.125-133
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• 2006

This study was conducted to detect the apoptosis incidence in blastocysts and to compare the abundance of Bax, Bcl2L1, VEGF and FGFR2 in in vitro fertilized (IVF), parthenogenetic (PAT) and nuclear transfer (NT) embryos. Oocytes matured for 40 hr were enucleated and reconstructed with confluenced fetal fibroblasts (FFs) derived from a ${\sim}45$ day fetus. Reconstructed eggs were then fused with 2 DC pulses (2.0 kV/cm, $30{\mu}sec$) and cultured with $7.5{\mu}g/ml$ cytochalasin B for 3 hr. Parthenotes (PAT) were produced with the same electric strength and culture for NT eggs. The embryos were cultured in NCSU-23 medium at $39^{\circ}C,\;5%\;CO_2,\;5%\l;O_2$ in air. In 3 runs, set of 10 embryos at the 4-cell to blastocyst stages were used to extract total RNA for analyzing the gene expression patterns of pro-apoptotic (Bax), anti-apoptotic (Bcl2L1), vasculogenesis (VEGF), implantation (FGFR2III) using real-time quantitative PCR. Cleavage and blastocyst rates were significantly higher (P<0.05) in IVF and PAT ($79.3{\pm}8.5\;and\;25.5{\pm}6.1,\;and\;85.0{\pm}6.4\;and\;38.6{\pm}5.5$, respectively)than NT counterparts ($65.1{\pm}5.2\;and\;15.6{\pm}3.0$, respectively). Significantly higher (P<0.05) total cells were observed in IVF controls and PAT ($34.7{\pm}5.8\;and\;38.1{\pm}4.1$) than NT embryos ($24.8{\pm}3.2$). Apoptosis index was significantly lower (P<0.05) in IVF than NT embryos. The Relative abundances (RA) of Bax and VEGF were significantly higher (P<0.05) at blastocyst stage in NT than IVF control. The RA of Bcl2L1 and FGFR2III were significantly higher (P<0.05) at blastocyst stage in IVF than NT. The present study observed the abnormal gene expressions in NT embryos at various developmental stages, suggesting certain clues to find out the cause of the low efficiency of NT to term.

• International Journal of Oral Biology
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• 제39권2호
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• pp.107-114
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• 2014

Taste is an important sense in survival and growth of animals. The growth and maintenance of taste buds, the receptor organs of taste sense, are under the regulation of various neurotrophic factors. But the distribution aspect of neurotrophic factors and their receptors in distinct taste cell types are not clearly known. The present research was designed to characterize mRNA expression pattern of neurotrophic factors and their receptors in distinct type of taste cells. In male 45-60 day-old Sprague-Dawley rats, epithelial tissues with and without circumvallate and folliate papillaes were dissected and homogenized, and mRNA expressions for neurotrophic factors and their receptors were determined by RT-PCR. The mRNA expressions of brain-derived neurotrophic factor (BDNF), neurotrophin-3 (NT3), receptor tyrosine kinase B (TrkB), exclusion of nerve growth factor (NGF), neurotrophin-4/5 (NT4/5), receptor tyrosine kinase A (TrkA), receptor tyrosine kinase C (TrkC), and p75NGFR were observed in some population of taste cell. In support of this result and to characterize which types of taste cells express NT3, BDNF, or TrkB, we examined mRNA expressions of NT3, BDNF, or TrkB in the $PLC{\beta}2$ (a marker of Type II cell)-and/or SNAP25 (a marker of Type III cell)-positive taste cells by a single taste cell RT-PCR and found that the ratio of positively stained cell numbers were 17.4, 6.5, 84.1, 70.3, and 1.4 % for $PLC{\beta}2$, SNAP25, NT3, BDNF, and TrkB, respectively. In addition, all of $PLC{\beta}2$-and SNAP25-positive taste cells expressed NT3 mRNA, except for one taste bud cell. The ratios of NT3 mRNA expressions were 100% and 91.7% in the SNAP25-and $PLC{\beta}2$-positive taste cells, respectively. However, two TrkB-positive taste cells co-expressed neither $PLC{\beta}2$ nor SNAP 25. The results suggest that the most of type II or type III cells express BDNF and NT3 mRNA, but the expression is shown to be less in type I taste cells.

• 한국식품위생안전성학회지
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• 제31권2호
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• pp.74-84
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• 2016

초등교사와 영양교사는 어린이 식생활 교육자로서 이들의 식품안전 및 위해에 대한 인식은 식생활교육에 큰 영향을 미칠 수 있다. 따라서 본 연구에서는 주요 식품 위해 요인으로 오인되고 있는 식품첨가물에 대해 수도권 소재 초등학교에 근무하는 초등교사(279명)와 영양교사(72명)를 대상으로 식품첨가물에 대한 인식 및 정보요구도에 대한 설문조사를 실시하여 분석함으로써, 식품첨가물에 대한 올바른 정보전달 촉진을 위한 기초자료를 확보하고자 하였다. 맛을 우선하는 교사(39.1%)에 비해 영양교사는 식품을 구입할 때 안전성(68.1%)을 가장 먼저 고려하여 유의한 차이(p < 0.001)를 보였고, 제조사, 가격 유통기한 외에 교사는 원산지(32.5%), 영양교사는 주성분(50.0%)을 우선 확인한다고 하였다(p < 0.01). 식품표시 이해도는, 주성분, 식품첨가물, 영양성분, 인증마크 등 모든 부분에서 영양교사가 높았으며, 교사(3.53), 영양교사(4.17) 모두 식품첨가물 관련 표시 이해도가 가장 낮게 나타났다. 교사, 영양교사 모두 식품의 안전을 위협하는 요인으로 환경오염물질, 식중독 세균과 바이러스, 식품첨가물 순서로 응답하였고, 가공식품 구매시 '무첨가', '무색소' 표시가 있는 식품을 구매한다는 응답 비율이 높았다. 또한 영양교사는 식품첨가물이 안전성, 유효성 평가를 거치고(100%), 사용기준 및 규격이 정해져 있다(81.9%)는 것을 안다고 응답했음에도 불구하고, 교사(75.2%)보다 많은 87.5%가 식품첨가물 섭취는 건강에 위험하다고 응답하는 괴리를 보여 이들에 대한 올바른 정보전달의 필요성을 확인할 수 있었다. 교사 및 영양교사 모두 식품첨가물에 대해 가장 알고싶어 하는 정보는 '안전성과 위험성'이었고, 정보매체로는 교사는 TV(3.80)를, 영양교사는 강의(3.65)를 가장 선호하였다. 식품첨가물에 대한 정보 제공기관이나 제공자에 대한 신뢰도에서 정부기관이나 식약처 공무원에 대한 신뢰도는 병원, 대학연구기관, 의사, 연구원 등에 대한 신뢰도 보다 낮았다. 대중매체의 정보는 신뢰하지 않는다는 비율이 높았으나, 대중매체를 접한 후 교사(39.4%)는 식품첨가물 표시를 보게 된다고 하였고, 영양교사(50%)는 식품첨가물이 함유된 식품을 줄인다고 응답하여 그 영향력을 확인할 수 있었다. 또한 대중매체 내용이 선정적 감성적 접근을 하여 문제가 있다고 답변하며, 정부가 잘못된 정보를 곧바로 바로잡을 수 있도록 전문가 집단을 활용하여 균형적으로 정보를 제공하여야 한다고 지적하였다. 본 연구 결과, 초등교사와 영양교사는 식품첨가물에 대해 각기 다른 인식과 정보요구도를 보이고 있어 어린이들에게 올바른 식생활교육을 할 수 있도록 식품첨가물에 대한 정보전달 노력이 요구된다.

• 한국수정란이식학회지
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• 제10권1호
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• pp.73-82
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• 1995

large scale production of cloned embryos requires the technology of multiple generation nuclear transplantation(NT) using NT embryos as the subsequent donor nuclei. The purposes of this study were producing the second generation cloned rabbit embryos, and also to determine the electrofusion rate and in vitro developmental potential comparatively in the cloned embryos of the first and second NT generation. The embryos of 16-cell stage were collected from the mated does by flushing oviducts with Dulbecco's phosphate buffered saline(D-PBS) containing 10% fetal calf serum(FCS) at 47 hours after hCG injection In the first generation NT, the nuclear donor embryos were synchronized in the phase of Gi /S transition of 32-cell stage. The first generation NT embryos which were developed to 8-cell were synchronized in Gi /S transition phase of the following 16-cell stage and used as donor nuclei for second generation Synchronization of the cell cycle of blastomeres was induced, first, using an inhibitor of microtuble polymerization, colcemid for 10 hours to arrest blastomeres in M phase, and secondly, using a DNA synthesis inhibitor, aphidicolin for 1.5 to 2 hours to arrest them in Gi /S transition boundary. The recipient cytoplasms were obtained by removing the nucleus and the first polar body from the oocytes collected at 14 hours after hCG injection. The separated donor blastomeres were injected into the enucleated recipient oocytes by micromanipulation and were electrofused by electrical stimulation of three pulses for 60 $\mu$sec at 1.25 kV /cm in 0.28 M rnannitol solution The fused oocytes were co-cultured with a monolayer of rabbit oviductal epithelial cells in M-199 solution containing 10% FCS for 120 hours at 39$^{\circ}C$ in a 5% $CO_2$ incubator. Following in vitro culture of the first and second generation cloned embryos to blastocyst stage, they were stained with Hoechst 33342 dye for counting the number of blastomeres by fluorescence microscopy. The results obtained were summarized as follows: 1. The electrofusion rate was found to be similar as 79.4 and 91.5% in the first and second generation NT rabbit embryos, respectively. 2. The in vitro developmental potential to blastocyst stage of the second generation NT embryos (23.3%) was found significantly(p<0.05) lower, compared with that of the first generation NT embryos (56.8%). 3. The mean blastomeres counts of embryos developed to blastosyst stage following in vitro culture for 120 hours and also their daily cell cycles during the culture period were decreased significantly (p<0.05) to 104.3 cells and 1.33 cylces in the second NT generation, compoared with 210.4 cells and 1.54 cycles in the first NT generation, respectively.

• 한국재료학회지
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• 제14권11호
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• pp.764-768
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• 2004

Perovskite Pb(Mn_{1/3}Sbu_{2/3})O_2-Pb(Zr,Ti)O_3\;(PMS-PZT) was prepared and ZnO doping effects on its piezoelectric properties were investigated. Pyrochlore phase was not identified in the PMS-PZT ceramics with $0\sim5\;mol\%$ ZnO sintered at $1100^{\circ}C$ for 2 hrs, and maximum sintered density of $7.92 g/cm^3$ was obtained. Piezoelectric charge constant and voltage constant increased to $359{\times}10^{-12}\;C/N\;and\;22.5{\times}10^{-13}\;Vm/N$, respectively, with increasing ZnO content. Mechanical quality factor reduced considerably with increasing ZnO content. When the ZnO content was 3 $mol\%$, electromechanical coupling factor and relative dielectric constant showed maximum values of $56\%$ and 1727, respectively. This should be evaluated by complicated variations of sintered density, tetragonality of lattice, grain size, and A-site vacancy generated by ZnO addition and $Zn^{2+}$ substitution.