• Journal of Embryo Transfer
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• v.30 no.3
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• pp.219-224
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• 2015

The purpose of this study is to develop transgenic cell line expressing targeted human granulocyte colony stimulating factor (hGCSF) and green fluorescence protein (GFP) genes as well as production of Somatic Cell Nuclear Transfer (SCNT) embryos derived from co-expressed transgenic donor cells. Constructed pPiggy-mWAP-hGCSF-EF1-GFP vector was chemically transfected into bovine fetus cells and then, only GFP expressed cells were selected as donor cells for SCNT. Cleavage and blastocyst rates of parthenogenetic, SCNT embryos using non-TG cell and hGCSF-GFP dual expressed SCNT embryos were examined (cleavage rate: $78.0{\pm}2.8$ vs. $73.1{\pm}3.2$ vs. $70.4{\pm}4.3%$, developmental rate: $27.2{\pm}3.2$ vs. $21.9{\pm}3.1$ vs. $17.0{\pm}2.9%$). Result indicated that cleavage and blastocyst rates of TG embryos were significantly lower (P<0.05) than those of parthenogenetic and non-TG embryos, respectively. In this study, we successfully produced hGCSF-GFP dual expressed SCNT embryos and cryopreserved to produce transgenic cattle for bioreactor system purpose. Further process of our research will transfer of transgenic embryos to recipients and production of hGCSF secreting cattle.

• Journal of Microbiology and Biotechnology
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• v.27 no.9
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• pp.1566-1575
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• 2017

MicroRNAs (miRNAs) are abundant in bovine milk and milk derived from other livestock, and they have functional roles in infants and in the secretion process of mammary glands. However, few studies have evaluated miRNAs in dairy processes, such as during cheese making and ripening. Thus, we investigated the characteristics of milk-derived miRNAs during the manufacturing and ripening of Camembert cheese as well as the microbiota present using the quantitative reverse transcription polymer chain reaction (RT-qPCR) and 16S rRNA pyrosequencing, respectively. Pyrosequencing showed that the cheese microbiota changed dramatically during cheese processing, including during the pasteurization, starter culture, and ripening stages. Our results indicated that the RNA contents per $200mg/200{\mu}l$ of the sample increased significantly during cheese-making and ripening. The inner cheese fractions had higher RNA contents than the surfaces after 12 and 22 days of ripening in a time-dependent manner (21.9 and 13.2 times higher in the inner and surface fractions than raw milk, respectively). We performed a comparative analysis of the miRNAs in each fraction by RT-qPCR. Large amounts of miRNAs (miR-93, miR-106a, miR-130, miR-155, miR-181a, and miR-223) correlated with immune responses and mammary glands were present in aged cheese, with the exception of miR-223, which was not present on the surface. Considerable amounts of miRNAs were also detected in whey, which is usually disposed of during the cheese-making process. Unexpectedly, there were no significant correlations between immune-related miRNAs and the microbial populations during cheese processing. Taken together, these results show that various functional miRNAs are present in cheese during its manufacture and that they are dramatically increased in amount in ripened Camembert cheese, with differences according to depth.

• Korean Journal of Animal Reproduction
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• v.15 no.1
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• pp.1-6
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• 1991

This study was carried out to investigate the effects of Ca, BSA, heparin, semen storage and individual bull on motility and acrosome reaction of bovine fresh sperm and sperm stored in lactose-egg yolk solution(LES) at 5$^{\circ}C$ for 4hours, and the results obtained were as follows: 1. When sperm was incubated in SCS containing Ca, BSA, Ca + BSA, heparin, heparin + Ca, heparin + BSA, and heparin + Ca + BSA for 15 minutes, there was significant difference in sperm motility among the treatments, especially BSA showed significantly higher sperm motility than the others. Also there was significant difference in sperm acrosome reaction among the treatments, especially BSA and Ca + BSA showed significantly higher sperm acrosome reaction than the others. 2. Bull KNC 1 showed significantly higher sperm motility than KNC 1, HOL 1 and 2 in both fresh and stored semen, however KNC 1 showed significantly lower sperm acrosome reaction than KNC 1, HOL 1 and 2. Therefore, there was significant difference in sperm motility and acrosome reaction among individual bulls. 3. When KNC 1 and KNC 2 sperm were incubated in SCS and SCS + Ca, SCS + BSA, SCS + Ca + BSA, SCS + heparin, SCS + heparin + Ca, SCS + heparin + BSA, and SCS + heparin + Ca + BSA, there was significant difference in sperm motility among individual bulls, especially BSA in KNC 1 and BSA, Ca and Ca + BSA in KNC 2 showedsignificantly higher motility than the others. However, there was significant difference in sperm acrosome reaction among individual bulls, Ca in KNC 1 and Ca + BSA in KNC 2 showed higher acrosome reaction than the others.

• Journal of Animal Science and Technology
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• v.46 no.5
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• pp.729-734
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• 2004

This study was performed to analyze the sequence variations of mtDNA D-loop and their effects on milk in Hanwoo(Korean cattle). The resulting sequences were compared with previously published sequences for other cattle breeds (GenBank JOI394). The Polymerase Chain Reaction was performed to amplify a total of 964 bp between nucleotide 15758 and 383 within D-loop region of mtDNA using specific primers. Twenty polymorphic sites by nucleotide substitution were found in mtDNA D-loop region of Hanwoo. The frequencies of positions at 8, 169, 16042, 16051, 16057, 16093, 16119, 16122, 16209, 16255 and 16302 nt with high levels of sequence polymorphism were 0.150, 0.950, 0.085, 0.138, 0.106, 0.085, 0.138, 0.212, 0.085, 0.148 and 0.180, respectively. The substitution effect at 16119(p<0.1) and 16185(p< 0.05) nt was found significant on milk production. Polymorphism of mtDNA sequence in D-Ioop region could be useful for the analysis of cytoplasmic genetic variation and associations with the other economically important traits and maternal lineage analysis in Hanwoo.

• Journal of Animal Science and Technology
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• v.52 no.6
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• pp.475-480
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• 2010

The bovine fatty acid binding protein 4 (FABP4) plays an important role to uptake intracellular fatty acid. It has been previously reported as a positional candidate gene for marbling score in livestock. The re-sequencing of FABP4 gene detected a polymorphic AT repeated sequence in intron II of FABP4 gene. Allelic distribution for this microsatellite marker was examined in other cattle breeds. A total of 8 alleles were detected with diverse repeat units (14 to 21 AT repeat) in Hanwoo and 7 breeds. Of the 8 alleles, the predominant alleles were $[AT]_{16}$, $[AT]_{18}$ and $[AT]_{19}$ in the Hanwoo and 7 cattle breeds. The linear mixed model for genotypic effect (3237AT) on carcass traits showed a significant effect on marbling score (MAR P=0.025) and live weight (LWT; P=0.04) in the 583 Hanwoo cattle population. Live weight (LW) was highest in the homozygous $(AT)_{17}$ genotype ($557.5{\pm}6.94$) and lowest in the heterozygous $(AT)_{16/17}$ genotype ($521.7{\pm}7.70$). On the other hand, the homozygous $(AT)_{17}$ genotype ($3.0{\pm}0.15$) has the highest effect on marbling score and the lowest effect was in homozygous (AT)$_{18}$ genotype ($2.2{\pm}0.15$). The marbling score difference between both groups was 0.8 which is around two times higher than SNP genotype effect on marbling score in Limousin $\times$ Wagyu crosses.

• The Korean Journal of Physiology and Pharmacology
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• v.16 no.6
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• pp.413-422
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• 2012

The purpose of this study is to investigated the mechanism of pharmacological action of local anesthetic and provide the basic information about the development of new effective local anesthetics. Fluorescent probe techniques were used to evaluate the effect of lidocaine HCl on the physical properties (transbilayer asymmetric lateral and rotational mobility, annular lipid fluidity and protein distribution) of synaptosomal plasma membrane vesicles (SPMV) isolated from bovine cerebral cortex, and liposomes of total lipids (SPMVTL) and phospholipids (SPMVPL) extracted from the SPMV. An experimental procedure was used based on selective quenching of 1,3-di(1-pyrenyl)propane (Py-3-Py) and 1,6-diphenyl-1,3,5-hexatriene (DPH) by trinitrophenyl groups, and radiationless energy transfer from the tryptophans of membrane proteins to Py-3-Py. Lidocaine HCl increased the bulk lateral and rotational mobility of neuronal and model membrane lipid bilayes, and had a greater fluidizing effect on the inner monolayer than the outer monolayer. Lidocaine HCl increased annular lipid fluidity in SPMV lipid bilayers. It also caused membrane proteins to cluster. The most important finding of this study is that there is far greater increase in annular lipid fluidity than that in lateral and rotational mobilities by lidocaine HCl. Lidocaine HCl alters the stereo or dynamics of the proteins in the lipid bilayers by combining with lipids, especially with the annular lipids. In conclusion, the present data suggest that lidocaine, in addition to its direct interaction with proteins, concurrently interacts with membrane lipids, fluidizing the membrane, and thus inducing conformational changes of proteins known to be intimately associated with membrane lipid.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.21 no.1
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• pp.739-744
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• 2020

This study evaluates the erosive potential and effects of mixed alcohols by analyzing the pH, titratable acidity, and fluorescence loss degree (△F). Following alcohol groups were investigated: Soju, Calamansi+soju, Yakult+soju, Cola+soju, and Energy drink+soju. The ratio of soju:beverage in the alcohol mixtures was 7:3. Ed. Notes: The sentence lacks clarity. Please review if the edit correctly portrays the meaning. If not, please revise appropriately. Measurement of the pH and titratable acidity (the amount of 1M NaoH solution required to raise to pH 5.5 (TA5.5) and 7.0 (TA7.0)) of alcohols was achieved by stirring with pH meter. The erosive effect of the alcohol mixtures on bovine tooth (△F) after 1, 2, 4, and 6 hours exposure were analyzed by quantitative light-induced fluorescence (QLF-D). All the mixed alcohols in this study showed an acidic pH, lower than 4.5. The average pH of mixed alcohols was 3.17 ± 0.50 whereas the pH of Soju was 8.6 ± 0.01. The TA5.5 and TA7.0 values of the mixed alcohols were 0.5~18 and 0.5~23.5, respectively. △F of the three tested mixed alcohol groups (except yakult+soju group) were observed to increase in a time-dependent manner. The calamansi mixed alcohol had the highest acidity potential and erosive effect among the tested groups. Taken together, the results indicate that the mixed alcohols have a strong erosive effect and potential on dental enamel.

• Restorative Dentistry and Endodontics
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• v.26 no.6
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• pp.451-463
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• 2001

목적 - Inflammatory cytokine으로 알려진 interleukin 1$\beta$, tumor necrosis factor $\alpha$는 치수 및 치근단질환에서 주요한 역할을 하며, 골흡수를 자극하고 골형성을 방해하는 것으로 알려져 왔다. 이들 cytokine은 주로 단핵세포/대식세포가 형성하는 것으로 알려져 왔으나 최근 연구에 의하면, PMN도 또한 이 런 cytokine들을 형성할 수 있다는 것이 보고되었다. 오랫동안 염증반응이나 면역반응에서 PMN의 역할이 주로 포식작용 을 통해 병원균을 제거하는 것이라고만 생각되어져 왔던 것을 생각하면, 새로운 발견이라 할 수 있다. 또, PMN은 IL-1ra도 생성하는 것으로 보고되었는데, IL-1ra란 IL-1의 생물학적 작용을 방해하는 인자이므로, IL-1과 밀접한 관련을 가지는 질환의 발전에 있어서 IL-1과 IL-1ra의 balance가 매우 중요한 역할을 할 것으로 생각된다 즉, IL-1ra는 IL-1$\beta$의 proinflammatory effect를 제한할 수 있는 negative feedback mechanism이라고 할 수 있다. 이 연구의 목적은 치수 및 치근단 조직의 감염에 있어서 주요 원인균인 Porphyromonas endodontalis의 LPS가 PMN의 IL-1$\beta$, TNF-$\alpha$, IL-1ra생성에 미치는 영향을 단백질과 mRNA 수준에서 관찰하는 것이다. 잘 알려진 non-oral bacterium인 E. coli의 LPS를 positive control로 사용하였으며, IL-1ra가 IL-1$\beta$의 생물학적 작용을 방해하는 작용을 관찰하기 위해, IL-1의 biological assay도 시행하였다. 방법 - P. endodontalis ATCC 35406을 혐기성 조건에서 배양하고, hot phenol-water extraction의 방법으로 LPS를 추출(crude LPS)한 후, 제조회사로부터 구입한 E. coli의 crude LPS와 함께 정제하였다. 건강한 자원자들을 대상으로 말초혈액을 채취한 후 dextran sedimentaion을 거쳐 Lymphoprep을 이용하여 PMN층을 분리하였다. 얻어진 세포들은 RPMI 1640 (supplemented with fetal bovine serum antibiotics)에 5$\times$$10^{6}$cells/ml이 되도록 resuspend시킨 후 각기 다른 농도 (0, 0.01, 0.1, 1 and 10$\mu$g/ml)의 LPS를 처리하여, 각기 다른 시간(Northern blot : 1, 2, 4시간 ELISA : 2, 6, 12, 18시간)동안 37$^{\circ}C$ in 5% $CO_2$ 의 조건으로 배양하였다. 상층액은 -7$0^{\circ}C$에 보관하였다가 추후에 ELISA를 이용한 단백질 농도 측정과 IL-1 biological assay에 사용되어졌으며, 배양된 세포로부터 RNA를 추출하여 Northern hybridization을 통해 mRNA expression을 관찰하였다. (중략)

• Polymer(Korea)
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• v.29 no.5
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• pp.468-474
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• 2005

PLGA and PCL copolymers initiated by carbitol as drug carriers were synthesized by ring-opening polymerization of L-lactide (LA), glycolide (GA), and $\varepsilon-caprolactone(\varepsilon-CL)$. Implantable wafers were simply fabricated by direct compression method after physical mixing of copolymers and bovine serum albumin-fluorescein isothiocyanate (BSA-FITC) as a model protein drug. The release amounts of BSA-FITC from wafers were determined by fluorescence intensity using the fluorescence spectrophotometer. Also, the release behavior of BSA-FITC on wafers was controlled by adding the additives such as collagen, small intestinal submucosa (SIS), poly(vinyl pyrrolidone) (PVP), and poly(thylene glycol) (PEG). The wafer prepared by PLGA and PCL exhibited slow release within $10\%$ for 30 days. But, those prepared by a variety of additives exhibited the controlled BSA release patterns with a dependence on the additive contents. furthermore, the wafers containing natural materials such as collagen and SIS showed more zero-order release profile than that with synthetic materials such as PVP and PEG. It was confirmed that the release of BSA from implantable wafers could be easily controlled by adding natural additives.

• Journal of Chest Surgery
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• v.25 no.5
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• pp.494-503
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• 1992

The durability of the xenograft cardiac substitute valves is of a great concern on the clinical grounds. Four groups of tc tal and consecutive patients to the end of study operated on between 1976 and 1984 were Group ISM, 291 patients of MVR, ISA, 65 patients of AVR, and ISMA, 107 patients of MVR+AVR with the standard Ionescu-Shiley bovine pericardial valve, and H, 163 patients of valve replacement with the Hancock porcine aortic vlave. Operative mortality was 5.2%[ISM], 10.8%[ISA], 7.5%[ISMA] and 6.1%[H]. Early survivors were followed up for a total of 1148.3 patient-years[pt-yrs] [ISM], 271.2 pt-yrs [ISA], 488.1 pt-yrs[ISMA] and 822.9 pt-yrs[H]. Linearized late mortality was 2.1% /pt-yr [ISM], 1.l%/pt-yr[ISA], 1.8%/pt-yr[ISMA] and 1.8% /pt-yr[H]. Thromboembolic complication was experienced at the linearized rate of 1.045% /pt-yr [ISM], 1.475%/pt-yr[ISA], 0.615%/pt-yr[ISMA] and 1.822%/pt-yr[H], and bleeding complication at the rate of 0.871% /pt-yr[ISM], 0.63% /pt-yr[ISA], 0.205% /pt-yr [ISMA] and 0.729%a /pt-yr[H], respectively. Prosthetic valve endocarditis occurred at the rate of 0.610% /pt-yr[ISM], 1.475% /pt-yr[ISA], 1.639% /pt-yr[ISMA] and 0.972% /pt-yr[H]. The linearized annual incidence of primary tissue failure was 1.655%/pt-yr[ISM], l. 475%/pt-yr[ISA], 1.639% /pt-yr[ISMA], 2.187% /pt-yr[H] and 1.785% /pt-yr[Group HM : MVR with Hancock valve]. The incidence of tissue failure was significantly high in the patients younger than 30 years of age compared with the older patients. The actuarial survival was 87.7$\pm$2.5% at 10 years[ISM], 94.3$\pm$3.2% at 11 years[ISA], 89.6$\pm$3.4% at 10 years[ISMA] and 81.3$\pm$6.6% at 12 years[HM], The freedom from thromboembolism was 93.2$\pm$2.0% at 10 years[ISM], 90.6$\pm$4.6% at 11 years[ISA], 95.8$\pm$2.6% at 10 years[ISMA] and 80.9$\pm$11.1% at 12 years[HM], And, the freedom from primary tissue failure was 84.2$\pm$3.8% and 28.1$\pm$23.0% at 9 and 10 years[ISM], 60.4$\pm$16.9% at 11 years[ISA], 62.3$\pm$12.7 at 10 years[ISMA] and 65.6$\pm$9.8% at 12 years[HM]. In conclusion, the standard Ionescu-Shiley and the Hancock bioprosthetic valves are excellent in their antithrombogenicity and long-term survival. However, the features of the structural failure with the prolonged follow-up beyond 10 years appear to be guarding, and the clinical indications of these bioprostheses seem to be quite limited.