• Journal of Embryo Transfer
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• v.32 no.4
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• pp.287-295
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• 2017

Mitochondrial dysfunction is found in oocytes and transmitted to offspring due to maternal obesity. Treatment of obese mothers with endoplasmic reticulum (ER) stress inhibitors such as salubrinal (SAL) can reverse the mitochondrial dysfunction and result in normal embryonic development. Pig oocytes have also shown ER stress mostly in metaphase II stage. ER stress in oocytes may hinder the in vitro production of pig embryos. This study investigated the effect of ER stress inhibition by SAL treatment during in vitro maturation (IVM) of porcine oocytes at 1, 10, 50 and 100 nM concentrations. Firstly, we tested various concentrations of SAL. SAL at 10 nM showed higher (P < 0.05) developmental competence to the blastocyst stage (55.6%) after parthenogenesis (PA) than control (44.2%) while not different from other concentrations (49.2, 51.6, and 50.8% for 1, 50, and 100 nM, respectively). Secondly, we performed time-dependent treatment at 10 nM of SAL for IVM of oocytes. It revealed that treatment with SAL during 22 to 44 h of IVM significantly improved PA embryonic development to the blastocyst stage compared to control (40.5, 46.3, 51.7 and 60.2% for control, 0 to 22 h, 22 to 44 h and 0 to 44 h of IVM, respectively, P < 0.05). Glutathione (GSH) content is an indicator of cytoplasmic maturation of oocytes. Reactive oxygen species (ROS) have a harmful effect on developmental competence of oocytes. For this, we determined the intraoocyte levels of GSH and ROS after 44 h of IVM. It was found that SAL increased intraoocyte GSH level and also decreased ROS level (P < 0.05). Finally, we performed somatic cell nuclear transfer (SCNT) after treating oocytes with 10 nM SAL during IVM. SAL treatment significantly improved blastocyst formation of SCNT embryos compared to control (39.6% vs. 24.7%, P < 0.05). Our results indicate that treatment of pig oocytes with ER stress inhibitor SAL during IVM improves preimplantation development PA and cloned pig embryos by influencing cytoplasmic maturation in terms of increased GSH content and decreased ROS level in IVM pig oocytes.

• Composites Research
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• v.33 no.2
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• pp.39-43
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• 2020

Recently, fabrication process of thermoplastic polyamide-based composites with recyclability as well as impact, chemical, and abrasion resistance have been widely studied. In particular, thermoplastic reactive resin transfer molding (TRTM) in which monomer with low viscosity is injected and in-situ polymerized inside mold has received a great attention, because thermoplastic melts are hard to impregnate fiber preform due to their very high viscosity. However, it is difficult to optimize the processing conditions because of high reactivity and sensitivity to external environments of the used monomer, ε-caprolactam. In this study, viscosity as an important process parameter in TRTM was measured during in-situ anionic polymerization of ε-caprolactam and the solutions for problems caused by high polymerization rate and sensitivity to moisture and oxygen were suggested. Reliability of the improved measurement technique was verified by comparing the viscosity behavior at various environmental conditions including humidity and atmosphere, and it is expected to be helpful for optimization of TRTM process.

• Membrane Journal
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• v.15 no.3
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• pp.224-232
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• 2005

The use of the lung assist device (LAD) would be well suited for acute respiratory failure (ARF) patients, combining the simplicity of mechanical ventilation with the ability of extracoporeal membrane oxygenators (ECMO) to provide temporary relief for the natural lungs. This study's specific attention was focused on the effect of membrane vibration in the LAD. Quantitative experimental measurements were performed to evaluate the performance of the device, and to identify membrane vibration dependence on blood hemolysis. We tried to decide upon excited frequency band of limit hemolysis when blood hemolysis came to through a membrane vibration action. The excited frequency of the module type 5, consisted of 675 hollow fiber membranes, showed the maximum gas transfer rate. We concluded that the maximum oxygen transfer rate seemed to be caused by the occurrence of maximum amplitude and the transfer of vibration to hollow fiber membranes. It was excited up to $25{\pm}5$ Hz at each blood flow rate of module type 5. We found that this frequency became the 2nd mode resonance riequency of the flexible in blood flow. Blood hemolysis was low at the excited frequency of $25{\pm}5$ Hz. Therefore, we decided that limit hemolysis frequency of this LAD was $25{\pm}5$ Hz.

• Journal of Microbiology and Biotechnology
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• v.25 no.8
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• pp.1339-1348
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• 2015

Until now, considerable effort has been made to engineer novel nitrogen-fixing organisms through the transfer of nif genes from various diazotrophs to non-nitrogen fixers; however, regulatory coupling of the heterologous nif genes with the regulatory system of the new host is still not well understood. In this work, a 49 kb nitrogen fixation island from P. stutzeri A1501 was transferred into E. coli using a novel and efficient transformation strategy, and a series of recombinant nitrogen-fixing E. coli strains were obtained. We found that the nitrogenase activity of the recombinant E. coli strain EN-01, similar to the parent strain P. stutzeri A1501, was dependent on external ammonia concentration, oxygen tension, and temperature. We further found that there existed a regulatory coupling between the E. coli general nitrogen regulatory system and the heterologous P. stutzeri nif island in the recombinant E. coli strain. We also provided evidence that the E. coli general nitrogen regulator GlnG protein was involved in the activation of the nif-specific regulator NifA via a direct interaction with the NifA promoter. To the best of our knowledge, this work plays a groundbreaking role in increasing understanding of the regulatory coupling of the heterologous nitrogen fixation system with the regulatory system of the recipient host. Furthermore, it will shed light on the structure and functional integrity of the nif island and will be useful for the construction of novel and more robust nitrogen-fixing organisms through biosynthetic engineering.

• Journal of Embryo Transfer
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• v.32 no.3
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• pp.87-94
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• 2017

Tissue engineering (TE) has been developed to create functional organs and tissue by combining 3D matrix and cells in vitro. Vascularization and angiogenesis are utmost important for supply of nutrients and oxygen in tissue engineered organs. The present study was performed to isolate and characterize primary endothelial cells (EC) from aorta of alpha 1, 3-enzyme galactosyltransferase knock out (GalT KO) pig, to minimize immune rejection and analyze body immune system for future xenotransplantation studies. Isolation of primary EC from aorta were performed by incubation with dispase for 8-10 min at $37^{\circ}C$. Primary EC were cultured in EC growth medium on different extra cellular matrix (ECM), either collagen or gelation. Primary EC exhibits morphological characteristics and showed positive expressions of EC specific marker proteins i.e. PECAM1, KDR and VWF despite of their ECM surface; however, on collagen based surface they showed increase in mRNA level analyzed by qPCR. Primary EC cultured on collagen were sorted by flow cytometer using KDR marker and cultured as KDR positive cells and KDR negative cells, respectively. KDR positive cells showed dramatically increased in PECAM1 and VWF level as compared to KDR negative cells. Based on the above results, primary EC derived from GalT KO are successfully isolated and survived continuously in culture without becoming overgrown by fibroblast. Therefore, they can be utilize for xeno organ transfer, tissue engineering, and immune rejection study in future.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.35 no.5
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• pp.469-475
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• 2002

The rotating biological contactor (RBC) was tested for treatment of aquacultural water in a pilot-scale recirculating aquaculture system. Performance of RBC on the treatment of nitrogen source such as total ammonia nitrogen (TAN), nitrite nitrogen and nitrate nitrogen and chemical oxygen demand (CODcr.) was evaluated. A system was stocked with nile tilapia at an initial rearing densities of $5\%$ and $7\%$ over 30 days. As increasing rearing density from $5\%$ to $7\%$, the TAN removal rates was increased from $39.4 g/m^3{\cdot}day$ to $86.0 g/m^3{\cdot}day$. But TAN removal efficiency was decreased from $24.5\%$ to $16.0\%$. The removal rate of $COD_Cr$ was higher than TAN. The RBC as an aerator was also evaluated for increasing dissolved oxygen concentration. For $5\%$ and $7\%$ of rearing density, the average aeration rate were $280 g/m^3{\cdot}day$ and $255 g/m^3{\cdot}day$, respectively.

• Journal of Embryo Transfer
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• v.22 no.3
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• pp.155-160
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• 2007

During in vitro culture of mammalian oocytes and embryos, the cells are exposed to the risks that cause cell injury or death. Numerous studies have been reported that the cell injury may be induced by the action of free radicals generated by auto-oxidation. This study was undertaken to investigate the optimal culture condition system for in vitro culture of porcine embryos. We first evaluated the effect of culture media on the porcine embryo development. NCSU-23 and PZM-5, culture medium tested, were failed to produce significant difference on the rate of blastocyst formation. In NCSU-23, the developmental rate was slightly higher than that in PZM-5. During in vitro maturation (IVM), fertilizaton (IVF), and culture (IVC) under 5 or 20% oxygen ($O_2$), the rates of cleavage and development were insignificantly different from each other under our culture condition (20% $O_2$, in NCSU-23), the mean cell number per blastocyst was $40{\pm}10$. These results showed that medium and $O_2$ concentration had no significant effect on the development of porcine embryos.

• Microbiology and Biotechnology Letters
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• v.48 no.3
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• pp.344-357
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• 2020

Strain improvement and bioprocess development were undertaken to enhance hyaluronic acid(HA) production by Streptococcus zooepidemicus cells. Using a high-yielding mutant strain, statistical medium optimization was carried out in shake flask cultures, resulting in 52% increase in HA production (5.38 g/l) at the optimal medium composition relative to the parallel control cultures. For sufficient supply of dissolved oxygen (DO), which turned out to be crucial for enhanced production of HA, agitation system and speed were intensively investigated in 5 L bioreactor cultures. Increase in oxygen mass transfer coefficient (k_La) through increment of agitation speed (rpm) and 35% expansion of diameter of the newly-designed impellers showed significantly positive effects on HA production. By installing an expanded Rushton-turbine impeller for efficient break-down of sparged air, and an extended marine impeller above the Rushton-turbine impeller for efficient mixing of the air-born viscous fermentation broth, maximum amount of HA (9.79 g/l) was obtained at 450 rpm, 1.8 times higher level than that of the corresponding flask culture. Subsequently, the possibility of bioprocess scale-up to a 50 L bioreactor was investigated. Despite almost identical maximum HA production (9.11 vs 9.25 g/l), the average HA volumetric productivity (r_p) of the 50 L culture turned out only 74% compared to the corresponding 5 L culture during the exponential phase, possibly caused by shear damages imposed on the producing cells at the high stirring in the 50 L culture. The scale-up process could be successfully achieved if a scale-up criterion of constant oxygen mass transfer coefficient (k_La) is applied to the 50 L pilot-scale bioreactor system.

• The Korean Journal of Physiology and Pharmacology
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• v.21 no.4
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• pp.377-384
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• 2017

Activation of protein kinase C (PKC) is closely linked with endothelial dysfunction. However, the effect of $PKC{\beta}II$ on endothelial dysfunction has not been characterized in cultured endothelial cells. Here, using adenoviral $PKC{\beta}II$ gene transfer and pharmacological inhibitors, the role of $PKC{\beta}II$ on endothelial dysfucntion was investigated in cultured endothelial cells. Phorbol 12-myristate 13-acetate (PMA) increased reactive oxygen species (ROS), p66shc phosphorylation, intracellular adhesion molecule-1, and monocyte adhesion, which were inhibited by $PKC{\beta}i$ (10 nM), a selective inhibitor of $PKC{\beta}II$. PMA increased the phosphorylation of CREB and manganese superoxide dismutase (MnSOD), which were also inhibited by $PKC{\beta}i$. Gene silencing of CREB inhibited PMA-induced MnSOD expression, suggesting that CREB plays a key role in MnSOD expression. Gene silencing of $PKC{\beta}II$ inhibited PMA-induced mitochondrial ROS, MnSOD, and ICAM-1 expression. In contrast, overexpression of $PKC{\beta}II$ using adenoviral $PKC{\beta}II$ increased mitochondrial ROS, MnSOD, ICAM-1, and p66shc phosphorylation in cultured endothelial cells. Finally, $PKC{\beta}II$-induced ICAM-1 expression was inhibited by Mito-TEMPO, a mitochondrial ROS scavenger, suggesting the involvement of mitochondrial ROS in PKC-induced vascular inflammation. Taken together, the results suggest that $PKC{\beta}II$ plays an important role in PMA-induced endothelial dysfunction, and that the inhibition of $PKC{\beta}II$-dependent p66shc signaling acts as a therapeutic target for vascular inflammatory diseases.

• Journal of Environmental Science International
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• v.19 no.8
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• pp.951-960
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• 2010

Sediment works as a resource for electric cells. This paper was designed in order to verify how sediment cells work with anodic material such as metal and carbon fiber. As known quite well, sediment under sea, rivers or streams provides a furbished environment for generating electrons via some electron transfer mechanism within specific microbial population or corrosive oxidation on the metal surfaces in the presence of oxygen or water molecules. We experimented with one type of sediment cell using different anodic material so as to attain prolonged, maximum electric power. Iron, Zinc, aluminum, copper, zinc/copper, and graphite felt were tested for anodes. Also, combined type of anodes-metal embedded in the graphite fiber matrix-was experimented for better performances. The results show that the combined type of anodes exhibited sustainable electricity production for ca. 600 h with max. $0.57\;W/m^2$ Al/Graphite. Meanwhile, graphite-only electrodes produced max. $0.11\;W/m^2$ along with quite stationary electric output, and for a zinc electrode, in which the electricity generated was not stable with time, therefore resulting in relatively sharp drop in that after 100 h or so, the maximum power density was $0.64\;W/m^2$. It was observed that the corrosive reaction rates in the metal electrodes might be varied, so that strength and stability in the electric performances(voltage and current density) could be affected by them. In addition to that, COD(chemical oxygen demand) of the sediment of the cell system was reduced by 17.5~36.7% in 600 h, which implied that the organic matter in the sediment would be partially converted into non-COD substances, that is, would suggest a way for decontamination of the aged, anaerobic sediment as well. The pH reduction for all electrodes could be a sign of organic acid production due to complicated chemical changes in the sediment.