• Asian-Australasian Journal of Animal Sciences
• /
• v.25 no.5
• /
• pp.621-628
• /
• 2012

The FAT-1 protein is an n-3 fatty acid desaturase, which can recognize a range of 18- and 20-carbon n-6 substrates and transform n-6 polyunsaturated fatty acids (PUFAs) into n-3 PUFAs while n-3 PUFAs have beneficial effect on human health. Fat1 gene is the coding sequence from Caenorhabditis elegans which might play an important role on lipometabolism. To reveal the function of fat1 gene in bovine fetal fibroblast cells and gain the best cell nuclear donor for transgenic bovines, the codon of fat1 sequence was optimized based on the codon usage frequency preference of bovine muscle protein, and directionally cloned into the eukaryotic expression vector pEF-GFP. After identifying by restrictive enzyme digests with AatII/XbaI and sequencing, the fusion plasmid pEF-GFP-fat1 was identified successfully. The pEF-GFP-fat1 vector was transfected into bovine fetal fibroblast cells mediated by Lipofectamine2000$^{TM}$. The positive bovine fetal fibroblast cells were selected by G418 and detected by RT-PCR. The results showed that a 1,234 bp transcription was amplified by reverse transcription PCR and the positive transgenic fat1 cell line was successfully established. Then the expression level of fat1 gene in positive cells was detected using quantitative PCR, and the catalysis efficiency was detected by gas chromatography. The results demonstrated that the catalysis efficiency of fat1 was significantly high, which can improve the total PUFAs rich in EPA, DHA and DPA. Construction and expression of pEF-GFP-fat1 vector should be helpful for further understanding the mechanism of regulation of fat1 in vitro. It could also be the first step in the production of fat1 transgenic cattle.

• Asian Pacific Journal of Cancer Prevention
• /
• v.15 no.6
• /
• pp.2753-2758
• /
• 2014

Background: GC-binding factor 2 (GCF2) is a transcriptional regulator that represses transcriptional activity of the epidermal growth factor receptor (EGFR) by binding to a specific GC-rich sequence in the EGFR gene promoter. In addition to this function, GCF2 has also been identified as a tumor-associated antigen and regarded as a potentially valuable serum biomarker for early human hepatocellular carcinoma (HCC) diagnosis. GCF2 is high expressed in most HCC tissues and cell lines including HepG2. This study focused on the influence of GCF2 on cell proliferation and apoptosis in HepG2 cells. Materials and Methods: GCF2 expression at both mRNA and protein levels in HepG2 cells was detected with reverse transcription (RT) PCR and Western blotting, respectively. RNA interference (RNAi) technology was used to knock down GCF2 mRNA and protein expression. Afterwards, cell viability was analyzed with a Cell Counting Kit-8 (CCK-8), and cell apoptosis and caspase 3 activity by flow cytometry and with a Caspase 3 Activity Kit, respectively. Results: Specific down-regulation of GCF2 expression caused cell growth inhibition, and increased apoptosis and caspase 3 activity in HepG2 cells. Conclusions: These primary results suggest that GCF2 may influence cell proliferation and apoptosis in HepG2 cells, and also provides a molecular basis for further investigation into the possible mechanism at proliferation and apoptosis in HCC.

• Journal of Nutrition and Health
• /
• v.24 no.4
• /
• pp.356-365
• /
• 1991

$L-Triiodothyronine(T_3)$ binding to purified plasma membrane and to isolated nuclei from the same liver in obese(ob/ob) mice and their lean littermates was examined. The maximal binding capacity(Bmax) for $T_3$ receptor of liver nuclei, as compared to lean control, was significantly lower in the obese mouse$(obese 527{\pm}80fmol/mg\;DNA ; lean 883{\pm}62fmol/mg\;DNA)$, without an apparent difference in dissociation constant(Kd). The finding that obese mice have fewer liver nuclear $T_3$ receptors confirms previous reports. The Bmax and Kd of liver plasma membrane $T_3$ receptor were not significantly different between obese and lean mouse, which suggests no defect to be occurring in the function of the plasma membrane $T_3$ receptor and reinforces the view that the peripherally impaired thyroid hormone action in obese mice is a post plasma membrane receptor event. These results further support the hypothesis that the major defect of the thyroid hormone metabolism in genetic obesity occurs at the level of the nuclear receptor.

• Clinical and Experimental Reproductive Medicine
• /
• v.38 no.4
• /
• pp.193-202
• /
• 2011

Objective: We found previously that $interferon$ $regulatory$ factor ($Irf$)-1 is a germinal vesicle (GV)-selective gene that highly expressed in GV as compared to metaphase II oocytes. To our knowledge, the function of $Irf-1$ in oocytes has yet to be examined. The present study was conducted to determine the relationship between retinoic acid (RA) and RA-mediated expression of $Irf-1$ and the mouse oocyte maturation. Methods: Immature cumulus-oocyte-complexes (COCs) were collected from 17-day-old female mice and cultured $in$ $vitro$ for 16 hours in the presence of varying concentrations of RA (0-10 ${\mu}M$). Rate of oocyte maturation and activation was measured. Gene expression was measured by quantitative real-time reverse transcription-polymerase chain reaction (RT-PCR) and cytokine secretion in the medium was measured by Bio-Plex analysis. Apoptosis was analyzed by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay. Results: The rates of oocyte maturation to metaphase II and oocyte activation increased significantly with RA treatment (10 nM-1 ${\mu}M$). With 100 nM RA treatment, lowest level of $Irf-1$ mRNA and cumulus cell's apoptosis was found. Among 23 cytokines measured by Bio-Plex system, the substantial changes in secretion of tumor necrosis factor-${\alpha}$, macrophage inflammatory protein-$1{\beta}$, eotaxin and interleukin-12 (p40) from COCs in response to RA were detected. Conclusion: We concluded that the maturation of oocytes and $Irf-1$ expression are negatively correlated, and RA enhances the developmental competence of mouse immature oocytes $in$ $vitro$ by suppressing apoptosis of cumulus cells. Using a mouse model, results of the present study provide insights into improved culture conditions for $in$ $vitro$ oocyte maturation and relevant cytokine production and secretion in assisted reproductive technology.

• Journal of Environmental Science International
• /
• v.19 no.11
• /
• pp.1423-1436
• /
• 2010

The effects of meteorological and reclaiming conditions on the reduction of suspended particles are investigated using a computational fluid dynamics (CFD) model with the k-$\varepsilon$ turbulence closure scheme based on the renormalization group (RNG) theory. Twelve numerical experiments with different meteorological and reclaiming conditions are performed. For identifying the meteorological characteristics of the target area and providing the inflow conditions of the CFD model, the observed data from the automatic weather station (AWS) near the target area is analyzed. Complicated flow patterns such as flow distortion, horse-shoe vortex, recirculation zone, and channeling flow appeared due to the topography and buildings in the domain. Specially, the flow characteristics around the reclamation area are affected by the reclaiming height, reclaiming size and windbreak height. Reclaiming height affected the wind speed above the reclaiming area. Windbreak induces more complicated flow patterns around the reclaiming area as well as within the reclaiming area. In front of the windbreak, flow is distorted as it impinges on the windbreak. As a result, upward flow is generated there. Behind the windbreak, a secondary circulation, so called, a recirculation zone is generated and flow is reattached at the end of the recirculation zone (reattachment point). At the lower part of the recirculation zone, there is a reverse flow toward the windbreak. Flow passing to the reattachment point starts to be recovered. Total amounts of suspended particles are calculated using the frictional and threshold frictional velocities, erosion potential function, and the number of surface disturbance. In the case of a 10 m-reclaiming and northerly wind, the amount of suspended particles is largest. In the presence of 5 m windbreak, the friction velocity above the reclaiming area is largely reduced. As a result, the total amount of the suspended particles largely decreases, compared to the case with the same reclaiming and meteorological conditions except for the windbreak The calculated suspended particle amounts are used as the emission rate of the dispersion model simulations and the dispersion characteristics of the suspended particles are analyzed.

• 한국지구물리탐사학회:학술대회논문집
• /
• 2004.08a
• /
• pp.25-48
• /
• 2004

Structures show the phehomena of deformation and lowering of function with time-lapse by artificial environments and changes of geotechnical conditions or accumulation of initial deformation elements. This study aims the structural assessment of cultural property, Chum-Sung-Dae, located in Kyeongju city, Korea. It was built about 1,300 years ago, and has undergone deformation and ground-subsidence with time-lapse. Non-destructive evaluation techniques were applied to the Chum-Sung-Dae, to protect it from survey Because of this reason, 3D precise laser scanning surveying system was applied to measure the exact size of Chum-Sung-Dae, displacement and declining angles. Geophysical exploration also was applied to study the subsurface distribution of geotechnical parameters or physical properties. Natural frequencies were measured from real and model of Chum-Sung-Dae to study the dynamic characteristics of vibration and/or earthquake load and stiffness of structures.

• Development and Reproduction
• /
• v.2 no.1
• /
• pp.21-27
• /
• 1998

The present study was performed to analyze the gene expressions of pituitary adenylate cyclase-activating polypeptide(PACAP) and its receptor in the rat uterus, a candidate for novel extrahypothalamic source and target. The PACAP cDNA fragments corresponding to the common exon region which is found in both the rat hypothalamus and testis were produced from all tissue samples including the rat uterus by reverse transcriptionpolymerase chain reaction (RT-PCR). No PCR product was amplified from the rat hypothalamic, pituitary, ovarian and uterine samples when the 5' primer corresponding to the testis-specific exon 1 region was used, while the predicted size of product was detected from the testis sample. RT-PCR using the uterine RNA and specific primers for the PACAP receptor yielded products with predicted sizes. Transcripts for the rat uterine PACAP receptor were identified as type I isoforms with hip-hop and hip- or hop-type inserts. After pregnant mare's serum gonadotropin (15 IU) treatment of immature rats (day 25), the level of PACAP mRNA was increased in 24 h and 48 h group, and was declined to the lowest in 72 h group. The present study shows the presence of transcripts for PACAP and its receptor isoform in the rat uterus. These finding ssuggest that the uterine PACAP ight act as a novel autocrine and/or paracrine factor via its specific receptors on the reglulation of rat uterine function and physiology during the reproductive cycle.

• Journal of the Institute of Electronics Engineers of Korea SD
• /
• v.38 no.10
• /
• pp.678-686
• /
• 2001

Origins for the transconductance dispersion and the gate leakage current in a GaAs metal semiconductor field effect transistor were found using capacitance deep-level transient spectroscopy (DLTS) measurements. In DLTS spectra, we observed two surface states with thermal activation energies of 0.65 $\times$ 0.07 eV and 0.88 $\times$ 0.04 eV and an electron trap EL2 with thermal activation energy of 0.84 $\times$ 0.01 eV. Transconductance was decreased in the frequency range of 5.5 Hz ~ 300 Hz. The transition frequency shifted to higher frequencies with the increase of temperature and the activation energy for the change of the transition frequency was determined to be 0.66 $\times$ 0.02 eV. From the measurements of the gate leakage current as a function of the device temperature, the forward and reverse currents are coincident with each other below gate voltages lower than 0.15 V, namely Ohmic behavior between gate and source/drain electrodes. The activation energy for the conductance of electrons on the surface of MESFET was 0.63 $\times$ 0.01 eV. Comparing activation energies obtained by different measurements, we found surface states H1 caused the transconductance dispersion and the fate leakage current.

• Asian Pacific Journal of Cancer Prevention
• /
• v.13 no.5
• /
• pp.2087-2093
• /
• 2012

Previous evidence showed ${\beta}1$, 3-N-acetylglucosaminyltransferase 8 (${\beta}3GnT8$), which can extend polylactosamine on N-glycans, to be highly expressed in some cancer cell lines and tissues, indicating roles in tumorigenesis. However, so far, the function of ${\beta}3GnT8$ in laryngeal carcinoma has not been characterized. To test any contribution, Hep-2 cells were stably transfected with sense or interference vectors to establish cell lines that overexpressed or were deficient in ${\beta}3GnT8$. Here we showed that cell proliferation was increased in ${\beta}3GnT8$ overexpressed cells but decreased in ${\beta}3GnT8$ knockdown cells using MTT. Furthermore, we demonstrated that change in ${\beta}3GnT8$ expression had significant effects on tumor growth in nude mice.We further provided data suggesting that overexpression of ${\beta}3GnT8$ enhanced the expression of matrix metalloproteinase-2 (MMP-2) and matrix metalloproteinase-9 (MMP-9) at both the mRNA and protein levels, associated with shedding of tissue inhibitors of metalloproteinase TIMP-2. In addition, it caused increased production of transforming growth factor beta 1 (TGF-${\beta}1$), whereas ${\beta}3GnT8$ gene knockdown caused the reverse effect. The results may indicate a novel mechanism by which effects of ${\beta}3GnT8$ in regulating cellular proliferation are mediated, at least in partvia targeting MMPs/TIMPs and TGF-${\beta}1$ in laryngeal carcinoma Hep-2 cells. The finding may lay a foundation for further investigations into the ${\beta}3GnT8$ as a potential target for therapy of laryngeal carcinoma.

• Asian-Australasian Journal of Animal Sciences
• /
• v.30 no.10
• /
• pp.1471-1477
• /
• 2017

Objective: Since athletic performance is a most importance trait in horses, most research focused on physiological and physical studies of horse athletic abilities. In contrast, the molecular analysis as well as the regulatory pathway studies remain insufficient for evaluation and prediction of horse athletic abilities. In our previous study, we identified AXL receptor tyrosine kinase (AXL) gene which was expressed as alternative spliced isoforms in skeletal muscle during exercise. In the present study, we validated two AXL alternative splicing transcripts (named as AXLa for long form and AXLb for short form) in equine skeletal muscle to gain insight(s) into the role of each alternative transcript during exercise. Methods: We validated two isoforms of AXL transcripts in horse tissues by reverse transcriptase polymerase chain reaction (RT-PCR), and then cloned the transcripts to confirm the alternative locus and its sequences. Additionally, we examined the expression patterns of AXLa and AXLb transcripts in horse tissues by quantitative RT-PCR (qRT-PCR). Results: Both of AXLa and AXLb transcripts were expressed in horse skeletal muscle and the expression levels were significantly increased after exercise. The sequencing analysis showed that there was an alternative splicing event at exon 11 between AXLa and AXLb transcripts. 3-dimentional (3D) prediction of the alternative protein structures revealed that the structural distance of the connective region between fibronectin type 3 (FN3) and immunoglobin (Ig) domain was different between two alternative isoforms. Conclusion: It is assumed that the expression patterns of AXLa and AXLb transcripts would be involved in regulation of exercise-induced stress in horse muscle possibly through an $NF-{\kappa}B$ signaling pathway. Further study is necessary to uncover biological function(s) and significance of the alternative splicing isoforms in race horse skeletal muscle.