• Asian-Australasian Journal of Animal Sciences
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• v.33 no.6
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• pp.1023-1033
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• 2020

Objective: The efficiency of the knock-in process is very important to successful gene editing in domestic animals. Recently, it was reported that transient loosening of the nucleosomal folding of transcriptionally inactive chromatin might have the potential to enhance homologous recombination efficiency. The objective of this study was to determine whether histone deacetylases (HDAC) inhibitor and RAD51 recombinase (RAD51) expression were associated with increased knock-in efficiency on the β-casein (bCSN2) gene locus in mammary alveolar-large T antigen (MAC-T) cells using the transcription activator-like effector nucleases (TALEN) system. Methods: MAC-T cells were treated with HDAC inhibitors, valproic acid, trichostatin A, or sodium butyrate for 24 h, then transfected with a knock-in vector, RAD51 expression vector and TALEN to target the bCSN2 gene. After 3 days of transfection, the knock-in efficiency was confirmed by polymerase chain reaction and DNA sequencing of the target site. Results: The level of HDAC 2 protein in MAC-T cells was decreased by treatment with HDAC inhibitors. The knock-in efficiency in MAC-T cells treated with HDAC inhibitors was higher than in cells not treated with inhibitors. However, the length of the homologous arm of the knock-in vector made no difference in the knock-in efficiency. Furthermore, DNA sequencing confirmed that the precision of the knock-in was more efficient in MAC-T cells treated with sodium butyrate. Conclusion: These results indicate that chromatin modification by HDAC inhibition and RAD51 expression enhanced the homologous recombination efficiency on the bCSN2 gene locus in MAC-T cells.

• Asian-Australasian Journal of Animal Sciences
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• v.27 no.3
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• pp.414-421
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• 2014

This study was conducted to determine the effects of saturated long-chain fatty acids (LCFA) on cell proliferation and triacylglycerol (TAG) content, as well as mRNA expression of ${\alpha}s1$-casein (CSN1S1) and genes associated with lipid and protein synthesis in bovine mammary epithelial cells (BMECs). Primary cells were isolated from the mammary glands of Holstein dairy cows, and were passaged twice. Then cells were cultured with different levels of palmitate or stearate (0, 200, 300, 400, 500, and 600 ${\mu}M$) for 48 h and fetal bovine serum in the culture solution was replaced with fatty acid-free BSA (1 g/L). The results showed that cell proliferation tended to be increased quadratically with increasing addition of stearate. Treatments with palmitate or stearate induced an increase in TAG contents at 0 to 600 ${\mu}M$ in a concentration-dependent manner, and the addition of 600 ${\mu}M$ was less effective in improving TAG accumulation. The expression of acetyl-coenzyme A carboxylase alpha, fatty acid synthase and fatty acid-binding protein 3 was inhibited when palmitate or stearate were added in culture medium, whereas cluster of differentiation 36 and CSN1S1 mRNA abundance was increased in a concentration-dependent manner. The mRNA expressions of peroxisome proliferator-activated receptor gamma, mammalian target of rapamycin and signal transducer and activator of transcription 5 with palmitate or stearate had no significant differences relative to the control. These results implied that certain concentrations of saturated LCFA could stimulate cell proliferation and the accumulation of TAG, whereas a reduction may occur with the addition of an overdose of saturated LCFA. Saturated LCFA could up-regulate CSN1S1 mRNA abundance, but further studies are necessary to elucidate the mechanism for regulating milk fat and protein synthesis.

• LOGISTICS
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• s.15
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• pp.52-57
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• 2006

2006 한국 국제 상용·특장차 전시회 3월 29일 개최/ 한솔CSN, 올 3천억 매출목표에 순조로운 출발/ (사)한국포장협회, 제4기 포장기술관리사 수료식 개최/ KLA, 아시아 최초 기내인터넷 장착기지 인증획득/ 유통업체들, "올해는 매출, 순익 두 마리 토끼잡는 해 될 것!"

• Development and Reproduction
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• v.8 no.1
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• pp.35-42
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• 2004

Oocyte maturation refers to the process that prophase I arrested germinal vesicle(GV) drives the progression of meiosis to metaphase II(MII) to have the capacity for fertilization and embryo development. To better understand the molecular mechanism(s) involved in oocyte maturation, we identified differentially expressed genes(DEGs) between GV and MII mouse oocytes using a new innovative annealing control primer (ACP) technology. Using 20 ACPs, we successfully cloned 32 DEGs between GV and Mll oocytes, and 26 out of these 32 DEGs were functionally known genes. Four genes including Pscd2 were GV-specific, 10 genes including PKD2 and CSN3 were highly expressed in GV oocytes(GV-selective), and 12 genes including Diva were highly expressed in MII oocytes (MII-selective). Ail of the genes identified in this study were first reported in the oocyte expression using ACP system and especially, we could characterize the existence of PKD-CSW signaling pathwayin the mouse oocytes. Results of the present study would provide insight for studying molecular mechanisms regulating oocyte maturation.

• Journal of Animal Science and Technology
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• v.63 no.5
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• pp.1126-1141
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• 2021

Recent evidence has shown that methionine (Met) supplementation can improve milk protein synthesis under hyperthermia (which reduces milk production). To explore the mechanism by which milk protein synthesis is affected by Met supplementation under hyperthermia, mammary alveolar (MAC-T) cells were incubated at a hyperthermic temperature of 42℃ for 6 h in media with different concentrations of Met. While the control group (CON) contained a normal amino acid concentration profile (60 ㎍/mL of Met), the three treatment groups were supplemented with Met at concentrations of 10 ㎍/mL (MET70, 70 ㎍/mL of Met), 20 ㎍/mL (MET80, 80 ㎍/mL of Met), and 30 ㎍/mL (MET90,90 ㎍/mL of Met). Our results show that additional Met supplementation increases the mRNA and protein levels of BCL2 (B-cell lymphoma-2, an anti-apoptosis agent), and decreases the mRNA and protein levels of BAX (Bcl-2-associated X protein, a pro-apoptosis agent), especially at an additional supplementary concentration of 20 ㎍/mL (group Met80). Supplementation with higher concentrations of Met decreased the mRNA levels of Caspase-3 and Caspase-9, and increased protein levels of heat shock protein (HSP70). The total protein levels of the mechanistic target of rapamycin (mTOR) and the mTOR signalling pathway-related proteins, AKT, ribosomal protein S6 kinase B1 (RPS6KB1), and ribosomal protein S6 (RPS6), increased with increasing Met supplementation, and peaked at 80 ㎍/mL Met (group Met80). In addition, we also found that additional Met supplementation upregulated the gene expression of αS1-casein (CSN1S1), β-casein (CSN2), and the amino acid transporter genes SLC38A2, SLC38A3 which are known to be mTOR targets. Additional Met supplementation, however, had no effect on the gene expression of κ-casein (CSN3) and solute carrier family 34 member 2 (SLC34A2). Our results suggest that additional Met supplementation with 20 ㎍/mL may promote the synthesis of milk proteins in bovine mammary epithelial cells under hyperthermia by inhibiting apoptosis, activating the AKT-mTOR-RPS6KB1 signalling pathway, and regulating the entry of amino acids into these cells.

• Asian-Australasian Journal of Animal Sciences
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• v.20 no.2
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• pp.166-171
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• 2007

The objective of this study was to assess the association of polymorphisms in ${\kappa}$-cn, ${\beta}$-lg, ${\beta}$-lg 5′ flanking region, CSN1S2, and IGFBP-3 genes with milk production traits and mastitis-related traits in Chinese Holstein. Traits analyzed were 305 day standard milk yield, protein percentage, fat percentage, the ratio of fat percentage and protein percentage, pre-somatic cell count, somatic cell count, and somatic cell score, respectively. CSN1S2 locus was uninformative because only one genotype BB was found in Chinese Holstein. Allele frequencies of A and B in IGFBP-3 gene were 0.5738 and 0.4262 in Chinese Holstein population, which was different from reported Qinchuan cattle population. The genotypes of animals at IGFBP-3 locus significantly affected 305 day standard milk yield, protein percentage, and somatic cell score. The ${\beta}$-lg genotypes had a significant effect on protein percentage and the ratio of fat percentage and protein percentage. Polymorphism in ${\beta}$-lg 5′ flanking region was associated with 305 day standard milk yield, protein percentage, fat percentage, pre-somatic cell count, and somatic cell count. No significant associations of the polymorphism in ${\kappa}$-cn gene were observed for any trait.

• Microbiology and Biotechnology Letters
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• v.32 no.1
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• pp.16-21
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• 2004

Six bacterial strains which formed large halo on chitosan-containing agar plate were isolated from beach mud and crabs at South coast of Korean peninsula. They were designated as Bacillus cereus KNUC51, B. cereus KNUC52, B. cereus KNUC53, B. cereus KNUC54, B. cereus KNUC55, and Paenibacillus favisporus KNUC56 by analysing their morphologies and 16S rDNA sequences. Chitosanase activities of all isolates were similar to that of B. subtilis 168. To enhance the activity of chitosanase, a powerful mutagen, MNNG was treated for P favisporus KNUC56. Three mutants showed higher activity of chitosanase than that of the original strain. The DNA fragments containing chitosanase gene from B. cereus sources were cloned, sequenced, and their deduced amino acid sequence analysis showed over 93% homologies with that of the known B. cereus ATCC14579. Extracellular sample from the isolates was incubated in proper reaction mixture including chitosan for 5 minutes at $37^{\circ}C$ to produce 3-10 chitosan oligomers which has been known to be active for clinical agents and agronomical agents.

• Journal of the Korean Institute of Intelligent Systems
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• v.14 no.5
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• pp.587-592
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• 2004

Context-aware computing systems require real-time context reasoning process for context awareness. Context reasoning can be done by comparing input information from sensors with knowledge-base within system. This method is identical with it of rule-based systems. In this paper, we propose hardware rule-based system architecture which can process context reasoning in real-time. Compared to previous architecture, hardware rule-based system architecture can reduce the number of constraints on rule representations and combinations of condition terms in rules. The modified content addressable memory, crossbar switch network and pre-processing module are used for reducing constraints. Using SystemC for description can provide easy modification of system configuration later.

• KSII Transactions on Internet and Information Systems (TIIS)
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• v.14 no.6
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• pp.2709-2734
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• 2020

The growing demand to make wireless data services 5G compatible has necessitated the development of an energy-efficient approach for an effective new wireless environment. In this paper, we first propose a cognitive sensor node (CSN) based game theory for deriving energy via a primary user-transmitted radio frequency signal. Cognitive users' time was segmented into three phases based on a time switching protocol: energy harvest, spectrum sensing and data transmission. The proposed model chooses the optimal energy-harvesting phase as the effected factor. We further propose a distributed energy-harvesting model as a utility function via pricing techniques. The model is a non-cooperative game where players can increase their net benefit in a selfish manner. Here, the price is described as a function pertaining to transmit power, which proves that the proposed energy harvest game includes Nash Equilibrium and is also unique. The best response algorithm is used to achieve the green connection between players. As a result, the results obtained from the proposed model and algorithm show the advantages as well as the effectiveness of the proposed study. Moreover, energy consumption was reduced significantly (12%) compared to the benchmark algorithm because the proposed algorithm succeeded in delivering energy in micro which is much better compared to previous studies. Considering the reduction and improvement in power consumption, we could say the proposed model is suitable for the next wireless environment represented in 5G.

• Advances in environmental research
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• v.9 no.2
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• pp.135-150
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• 2020

Many industries, such as textiles, chemical refineries, leather, plastics and paper, use different dyes in various process steps. At the same time, these industrial sectors are responsible for discharging contaminants that are harmful and toxic to humans and microorganisms by introducing synthetic dyes into wastewater. Of these dyes, methylene blue dye, which is classified as basic dyes, is accepted as a model dye. For this reason, methylene blue dye was selected in the study and its removal from the water was studied. In this study, two efficient biosorbents were developed from chitosan and sunflower waste, an agro-industrial waste and modified using iron nanoparticles. The biosorption efficiency was evaluated for methylene blue (MB) dye removal from aqueous solution under various parameters such as treating agent, solution pH, biosorbent dosage, contact time, initial dye concentration and temperature. We investigated the kinetic properties of dye removal from water for Chitosan-Sunflower (CS), Chitosan-Sunflower-Nanoiron (CSN). When the wavelength of MB dye was spectrophotometrically scanned, the maximum absorbance was determined as 660 nm. For the core shell biosorbents we obtained, we found that the optimum time for removal of MB from wastewater was 60 min. The pH of the best pH was determined as 5 in the studied pH. The most suitable temperature for the experiment was determined as 30℃. SEM-EDAX, TEM, XRD, and FTIR techniques were used to characterize biosorbents produced and modified in the experimental stage and to monitor the change of biosorbent after dye removal. The interactions of the paint with the surface used for removal were explained by these techniques. It was calculated that 80% of CS and 88% of CSN removed MB in optimum conditions. Also, the absorption of MB dye onto the surface was investigated by Langmiur and Frendlinch isotherms and it was determined from the results that the removal was more compatible with Langmiur isotherm.