• 한국수정란이식학회지
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• 제17권2호
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• pp.101-108
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• 2002

Human Prourokinase (proUK) offers potential as a novel agent with improved fibrin specificity and, as such, may offer advantages as an attractive alternative to urokinase that is associated with clinical benefits in patients with acute peripheral arterial occlusion. For production of transgenic cow as human proUK bioreacotor, we conducted this study to establish efficient production system for bovine transgenic embryos by somatic cell nuclear transfer (NT) using human prourokinase gene transfected donor cell. An expression plasmid for human prourokinase was constructed by inserting a bovine beta-casein promoter, a green fluorescent protein (GFP) marker gene, and human prourokinase target gene into a pcDNA3 plasmid. Cumulus cells were used as donor cell and transfected with the expression plasmid using the Fugene 6 as a carrier. To increase the efficiency for the production of transgenic NT, development rates were compared between non-transfected and transfected cell in experiment 1, and in experiment 2, development rates were compared according to level of GFP expression in donor cells. In experiment 1, development rates of non-transgenic NT embryos were significantly higher than transgenic NT embryos (43.3 vs. 28.4％). In experiment 2, there were no significant differences in fusion rates (85.4 vs. 78.9％) and cleavage rates (78.7 vs. 84.4％) between low and high expressed cells. However, development rates to blastocyst were higher in low expressed cells (17.0 vs. 33.3％), and GFP expression rates in blastocyst were higher in high expressed cells (75.0 vs. 43.3％), significantly.

• Journal of Nutrition and Health
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• 제33권7호
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• pp.697-702
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• 2000

The objective of this study was to determine the extent to which the compensatory nutrition regimen modulates lactation performance and milk protein gene expression in the first and second lactation cycles. Female rats(28 days of age) were assigned to 1)control ad libitum ; 2) stari-step compensatory nutrition(SSCN) regimen an alternating 3-2-3-4-week schedule beginning with an energy restriction diet(40% restriction) for 3 weeks followed by the control diet(ad libitum) for 2 weeks and then alternating another 3-4 week feeding regimen. The SSCN rats were received an overall 20% energy restriction(average from all stair-step periods) compared with the conventionally fed control group. Rats were bred during the first week of the second realimentation. All pups were weaned on day 21 of lactation. About 1 week after weaning all dams were mated for the second pregnancy. Mammary tissues were obtained from pregnant and lactating rats during the first and second lactation cycles. During these lactation cycles the SSCN group had a 11% increase in average lactation performance over that of control. The SSCN group had significantly increased levels of milk protein gene($\alpha$- and $\beta$-casein) expression in mammary tissues during the first lactation cycle compared with those of the control group. During the second lactation period the levels of milk protein gene expression in lactating mammary tissues of the SSCN group were also higher than those of the control group. These results suggest that the effects of compensatory growth imposed at an early age extend to the second lactation cycle with regard to increased lactation performance and milk protein gene expression.

• Asian Pacific Journal of Cancer Prevention
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• 제17권3호
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• pp.1163-1168
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• 2016

Cholangiocarcinoma (CCA) is the bile duct cancer which constitutes one of the important public health problems in Thailand with high mortality rate, especially in the Opisthorchis viverrini (a parasite risk factor for CCA) endemic area of the northeastern region of the country. This study aimed to identify potential biomarkers from the plasma peptidome by CCA patients. Peptides were isolated using 10 kDa cut-off filter column and the flow-through was then used as a peptidome for LC-MS/MS analysis. A total of 209 peptides were obtained. Among these, 15 peptides were concerned with signaling pathways and 12 related to metabolic, regulatory, and biosynthesis of secondary metabolite pathways. Five exclusive peptides were identified as potential biomarkers, i.e. ETS domain-containing transcription factor ERF (P50548), KIAA0220 (Q92617), phosphatidylinositol 4,5-bisphosphate 3-kinase catalytic subunit beta isoform isoform 1 (P42338), LP2209 (Q6XYC0), and casein kinase II subunit alpha (P19784). Three of these biomarkers are signaling related molecules. A combination of these biomarkers for CCA diagnosis is proposed.

• 한국수정란이식학회:학술대회논문집
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• 한국수정란이식학회 2002년도 국제심포지엄
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• pp.75-75
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• 2002

The present study was conducted for the production of transgenic cloned cows by somatic cell nuclear transfer (SCNT) that secrete human prourokinase into milk. To establish an efficient production system for bovine transgenic SCNT embryos, the offset was examined of various conditions of donor cells including cell type, size, and passage number on the developmental competence of transgenic SCNT embryos. An expression plasmid far human prourokinase (pbeta-ProU) was constructed by inserting a bovine beta-casein promoter, a green fluorescent protein (GFP) marker gene, and a human prourokinase target gene into a pcDNA3 plasmid. Three types of bovine somatic cells including two adult cells (cumulus cells and ear fibroblasts) and fetal fibroblasts were prepared and transfected using a lipid-meidated method. In Experiment 1, developmental competence and rates of GFP expression in bovine transgenic SCNT embryos reconstructed with cumulus cells were significantly higher than those from fetal and ear fibroblasts. In Experiment 2, the effect of cellular senescence in early (2 to 4) and late (8 to 12) passages was investigated. No significant differences in the development of transgenic SCNT embryos were observed. In Experient 3, different sizes of GFP-expressing transfected cumulus cells [large (>30 ${\mu}{\textrm}{m}$) or small cell (<30 ${\mu}{\textrm}{m}$)] were used for SCNT. A significant improvement in embryo development and GFP expression was observed when small cumulus cells were used for SCNT. Taken together, these results demonstrate that (1) adult somatic cells could serve as donor cells in transgenic SCNT embryo production and cumulus cells with small size at early passage were the optimal cell type, and (2) transgenic SCNT embryos derived from adult somatic cells have embryonic development potential.

• Journal of Microbiology and Biotechnology
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• 제17권9호
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• pp.1469-1476
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• 2007

Jeot-gal is a traditional Korean fermented seafood and has long been used for seasoning. We isolated 188 strains from shrimp, anchovy, and yellow corvina Jeot-gal, and screened sixteen strains that showed strong fibrinolytic activities on a fibrin plate. Among those strains, the strain that had the largest halo zone was chosen and identified as Bacillus licheniformis by using 16S rDNA sequencing and an API CHB kit. The fibrinolytic activity of Bacillus licheniformis was characterized and designated as bpKJ-31. The active component of bpKJ-31 was identified as a 37 kDa protein, designated bacillopeptidase F, by internal peptide mapping and N-terminal sequencing. The optimum activity of bpKJ-31 was shown at pH 9 and $40^{\circ}C$, with a chromogenic substrate for plasmin. It had high degrading activity for the $B{\beta}$-chain and $A{\alpha}$-chain of fibrin(ogen), and also acted on thrombin, but not skim milk and casein. The amidolytic activity of bpKJ-31 was inhibited by 1 mM phenylmethanesulfonyl fluoride, but 1 mM EDTA did not affect the enzyme activity, indicating that bpKJ-31 is an alkaline serine protease, like a plasmin. The bpKJ-31 showed approximately 14.3% higher fibrinolytic activity than the plasmin. These features of bpKJ-31 make it attractive as a health-promoting biomaterial.

• Journal of Dairy Science and Biotechnology
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• 제30권1호
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• pp.17-22
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• 2012

Since the prevalence of allergies is increasing, food allergy is a major concern for consumers, as well as for the food industry. The foods that account for over 90% of all moderate to severe allergic reactions to food are milk, eggs, peanuts, soybeans, fish, shellfish, wheat, and tree nuts. Of these food allergens, milk is one of the major animal food allergens in infants and young children. Milk is the first food that an infant is exposed to; therefore, the sensitization rate of milk in sensitive individuals is understandably higher. The mechanisms involved in allergic reactions caused by this hypersensitivity are similar to those of other immune-mediated allergic reactions. The reactions occur in the gastrointestinal tract, skin, and respiratory tract, with headaches and psychological disorders occurring in some instances. The major allergenic proteins in milk are casein, ${\beta}$-lactoglobulin, and ${\alpha}$-lactalbumin, while some of the minor allergenic proteins are lactoferrin, bovine serum albumin, and immunoglobulin. Reliable allergen detection and quantification are essential for compliance with food allergen-labeling regulations, which protect the consumer and facilitate international trade.

• 대한치과교정학회지
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• 제44권3호
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• pp.113-118
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• 2014

Objective: The purpose of this in vitro study was to examine the effects of fluoridated, casein phosphopeptide-amorphous calcium phosphate complex (CPP-ACP)-containing, and functionalized ${\beta}$-tricalcium phosphate (fTCP)-containing toothpastes on remineralization of white spot lesions (WSLs) by using Quantitative light-induced fluorescence (QLF-D) Biluminator$^{TM}$ 2. Methods: Forty-eight premolars, extracted for orthodontic reasons from 12 patients, with artificially induced WSLs were randomly and equally assigned to four treatment groups: fluoride (1,000 ppm), CPP-ACP, fTCP (with sodium fluoride), and control (deionized water) groups. Specimens were treated twice daily for 2 weeks and stored in saliva solution (1:1 mixture of artificial and human stimulated saliva) otherwise. QLF-D Biluminator$^{TM}$ 2 was used to measure changes in fluorescence, indicating alterations in the mineral contents of the WSLs, immediately before and after the 2 weeks of treatment. Results: Fluorescence greatly increased in the fTCP and CPP-ACP groups compared with the fluoride and control groups, which did not show significant differences. Conclusions: fTCP- and CPP-ACP-containing toothpastes seem to be more effective in reducing WSLs than 1,000-ppm fluoride-containing toothpastes.

• Archives of Pharmacal Research
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• 제29권5호
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• pp.424-429
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• 2006

In the present study, a human mammary epithelial cell (HMEC) culture model was developed to evaluate the potential involvement of carrier-mediated transport systems in drug transfer into milk. Trypsin-resistant HMECs were seeded on $Matrigel^{circledR}-coated$ filters to develop monolayers of functionally differentiated HMEC. Expression of the specific function of HMEC monolayers was dependent of the number of trypsin treatments. Among the monolayers with different numbers of treatment (treated 1 to 3 times), the monolayer treated 3 times (3-t-HMEC monolayer) showed the highest maximal transepithelial resistance and expression of $\beta-casein$ mRNA as an index of differentiation. Transport of tetraethylammonium (TEA) across the 3-t-HMEC monolayer in the basolateral-to-apical direction was significantly higher than that in the apical-to-basolateral direction (p<0.05), whereas such directionality was not observed for p-aminohippurate, suggesting the existence of organic cation transporters, but not organic anion transporters. In fact, expression of mRNAs of human organic cation transporter (OCT) 1 and 3 were detected in the 3-t-HMEC monolayer. These results indicate that the 3-t-HMEC monolayer is potentially useful for the evaluation of carrier-mediated secretion of drugs including organic cations into human milk.

• 생명과학회지
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• 제13권4호
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• pp.535-540
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• 2003

최근 발효식품의 건강학적 활용면에 관한 연구는 활발히 진행되고 있다. 또한 발효식품 중에 가장 중요한 간장 발효미생물의 건강제품의 활용에 대해서 많은 연구가 진행되었다. 재래식 간장에 관한 연구결과로 항암작용, 노화지연, 호르몬 분비 촉진 등이 보고되고 있으나, 구강질환 원인균에 대한 방제효과가 있는가에 관한 연구는 거의 이루어지지 않고 있는 실정이다. 본 연구는 치아우식 등 구강질환 원인균인 Streptococcus sanguis, S. salivarius, S. mutans들의 성장을 억제하는 방제균을 선발하고 이들을 동정하기 위해 시행되었다. 우선 25종의 간장발효 균주를 분리하였고, 그 중 구강질환 방제력이 높은 2 균주(NG 06, NG 16)를 선별하였다. Bergey's manual of systematic bacteriology의 세균분류동정표에 의해 각종 동정에 필요한 배양학적, 형태학적, 생화학적 특성을 시험한 결과와 $Biolog^{(R)}$사의 세균동정시스템(MicroLogTM 3)을 이용하여 검정 실험한 결과 NG 06은 Bacillus racemilacticus로 NG 16은 Bacillus amyloliquefaciens으로 최종 확인, 동정되었다(Table 3). 앞으로 이 균들이 생산해내는 항균물질의 길항 기작에 관한 연구와 그 효능을 검증하고자 하며 그 항균물질 효과생산 최적조건에 대한 연구를 수행할 계획이다. 나아가 세치제 및 구강위생용품으로 활용면에 관한 연구를 확대하고자 한다.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2001년도 춘계학술발표대회
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• pp.60-60
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• 2001

Human thrombopoietin (hTPO) is a cytokine that plays a central role in megakaryopoiesis by influencing on the development and maturation of megakaryocyte and platelet production. To induce hTPO production in the mammary gland, expression vector was constructed by combining the promoter of bovine beta-casein gene, cDNA of hTPO and neomycine resistance gene for transfection into fibroblasts. Bovine fibroblast cells derived from female ear skin were transfercted with the expression vector using Lipofectamine (Life Technology, NY). Transected cells resistant to G4l8 treatment (600 $\mu\textrm{g}$/$m\ell$) were recovered and colony formation was initiated at 13 days. The colonies with about 1 cm diameter were picked and analysed by PCR. Single transfected cells were individually transferred to enucleated oocytes. After electrofusion, the reconstructed embryos were exposed to calcium ionophore (5uM) for 5 min followed by treatment with 6-DMAP (2.5 mM) for 4h. The nuclear transfer embryos were cultured in CRlaa medium at 38.5C, 5% $CO_2$ for 7 days. Twenty three of 29 (79.3%) colonies were proved to be hTPO transfectants by PCR. The colonies were further passaged and used to produce transgenic embryos using nuclear transfer. Cleavage and developmental rates of reconstructed embryos to the blastocyst stage were 65.1% and 39.4%, respectively Of 22 blastocysts that developed from reconstructed embryos with the transfected cell, 20 embryos (90.9%) were positive for hTPO by using PCR analysis. The results suggest that somatic cell nuclear transfer is efficient for production of transgenic embryos.