• Animal Bioscience
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• v.37 no.3
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• pp.437-450
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• 2024

Objective: Vanin-1 (VNN1) is a pantetheinase that catalyses the hydrolysis of pantetheine to produce pantothenic acid and cysteamine. Our previous studies have shown that the VNN1 is specifically expressed in chicken liver which negatively regulated by microRNA-122. However, the functions of the VNN1 in lipid metabolism in chicken liver haven't been elucidated. Methods: First, we detected the VNN1 mRNA expression in 4-week chickens which were fasted 24 hours. Next, knocked out VNN1 via CRISPR/Cas9 system in the chicken Leghorn Male Hepatoma cell line. Detected the lipid deposition via oil red staining and analysis the content of triglycerides (TG), low-density lipoprotein-C (LDL-C), and high-density lipoprotein-C (HDL-C) after VNN1 knockout in Leghorn Male Hepatoma cell line. Then we captured various differentially expressed genes (DEGs) between VNN1-modified LMH cells and original LMH cells by RNA-seq. Results: Firstly, fasting-induced expression of VNN1. Meanwhile, we successfully used the CRISPR/Cas9 system to achieve targeted mutations of the VNN1 in the chicken LMH cell line. Moreover, the expression level of VNN1 mRNA in LMH-KO-VNN1 cells decreased compared with that in the wild-type LMH cells (p<0.0001). Compared with control, lipid deposition was decreased after knockout VNN1 via oil red staining, meanwhile, the contents of TG and LDL-C were significantly reduced, and the content of HDL-C was increased in LMH-KO-VNN1 cells. Transcriptome sequencing showed that there were 1,335 DEGs between LMH-KO-VNN1 cells and original LMH cells. Of these DEGs, 431 were upregulated, and 904 were downregulated. Gene ontology analyses of all DEGs showed that the lipid metabolism-related pathways, such as fatty acid biosynthesis and long-chain fatty acid biosynthesis, were enriched. KEGG pathway analyses showed that "lipid metabolism pathway", "energy metabolism", and "carbohydrate metabolism" were enriched. A total of 76 DEGs were involved in these pathways, of which 29 genes were upregulated (such as cytochrome P450 family 7 subfamily A member 1, ELOVL fatty acid elongase 2, and apolipoprotein A4) and 47 genes were downregulated (such as phosphoenolpyruvate carboxykinase 1) by VNN1 knockout in the LMH cells. Conclusion: These results suggest that VNN1 plays an important role in coordinating lipid metabolism in the chicken liver.

• Animal Bioscience
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• v.34 no.2
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• pp.172-184
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• 2021

Objective: Vanin1 (VNN1) is a pantetheinase that can catalyze the hydrolysis of pantetheine to produce pantothenic acid and cysteamine. Our previous studies showed that VNN1 is specifically expressed in chicken liver. In this study, we aimed to investigate the roles of peroxisome proliferators activated receptor α (PPARα) and miRNA-181a-5p in regulating VNN1 gene expression in chicken liver. Methods: 5'-RACE was performed to identify the transcription start site of chicken VNN1. JASPAR and TFSEARCH were used to analyze the potential transcription factor binding sites in the promoter region of chicken VNN1 and miRanda was used to search miRNA binding sites in 3' untranslated region (3'UTR) of chicken VNN1. We used a knock-down strategy to manipulate PPARα (or miRNA-181a-5p) expression levels in vitro to further investigate its effect on VNN1 gene transcription. Luciferase reporter assays were used to explore the specific regions of VNN1 targeted by PPARα and miRNA-181a-5p. Results: Sequence analysis of the VNN1 promoter region revealed several transcription factor-binding sites, including hepatocyte nuclear factor 1α (HNF1α), PPARα, and CCAAT/enhancer binding protein α. GW7647 (a specific agonist of PPARα) increased the expression level of VNN1 mRNA in chicken primary hepatocytes, whereas knockdown of PPARα with siRNA increased VNN1 mRNA expression. Moreover, the predicted PPARα-binding site was confirmed to be necessary for PPARα regulation of VNN1 gene expression. In addition, the VNN1 3'UTR contains a sequence that is completely complementary to nucleotides 1 to 7 of miRNA-181a-5p. Overexpression of miR-181a-5p significantly decreased the expression level of VNN1 mRNA. Conclusion: This study demonstrates that PPARα is an important transcriptional activator of VNN1 gene expression and that miRNA-181a-5p acts as a negative regulator of VNN1 expression in chicken hepatocytes.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.48 no.4
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• pp.403-410
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• 2015

Viral nervous necrosis (VNN) is a serious disease of sevenband grouper Epinephelus septemfasciatus in Korean aquaculture farms. However, we suggest the following preventative methods for hatcheries: 1) disinfecting rearing water, 2) selecting spawners via ELISA and PCR, 3) selecting eggs via PCR, 4) disinfecting fertilized eggs, and 5) proper facilities management. When these methods are implemented, nervous necrosis virus (NNV)-free fish are produced because vertical and horizontal transmission is prevented. However, horizontal transmission of NNV through rearing seawater sourced from the environment during grow-out stages in sea cages can still occur. Live NNV vaccines with a low rearing temperature or Poly(I:C) immunization are very effective at preventing horizontal transmission of NNV in rearing farms. Furthermore, even after VNN is contracted, fish mortality can be reduced by administering Poly(I:C).

• The Journal of Korea Institute of Information, Electronics, and Communication Technology
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• v.9 no.1
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• pp.120-131
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• 2016

In this paper, a 512b MTP memory IP is designed by using MTP memory cells which are written by the FN (Fowler-Nordheim) tunneling method with only MV (medium voltage) devices of 5V which uses the back-gate bias, that is VNN (negative voltage). The used MTP cell consists of a CG (control gate) capacitor, a TG (tunnel gate) transistor, and a select transistor. To reduce the size of the MTP memory cell, just two PWs (P-wells) are used: one for the TG and the select transistors; and the other for the CG capacitor. In addition, just one DNW (deep N-well) is used for the entire 512b memory cell array. VPP and VNN generators supplying pumping voltages of ${\pm}8V$ which are insensitive to PVT variations since VPP and VNN level detectors are designed by a regulated voltage, V1V (=1V), provided by a BGR voltage generator.

• The Journal of Korea Institute of Information, Electronics, and Communication Technology
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• v.9 no.4
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• pp.419-427
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• 2016

In this paper, a 64b small-area MTP memory IP is designed. A VPPL (=VPP/3) regulator and a VNN (=VNN/3) charge pump are removed since the inhibit voltages of an MTP memory cell are all 0V instead of the conventional voltages of VPP/3 and VNN/3. Also, a VPP charge pump is removed since the VPP program voltage is supplied from an external pad. Furthermore, a VNN charge pump is designed to provide its voltage of -VPP as a one-stage negative charge pump using the VPP voltage. The layout size of the designed 64b MTP memory IP with MagnaChip's $0.18{\mu}m$ BCD process is $377.585{\mu}m{\times}328.265{\mu}m$ (=0.124mm2). Its DC-DC converter related layout size is 76.4 percent smaller than its conventional counterpart.

• Journal of the Korea Institute of Information and Communication Engineering
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• v.14 no.8
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• pp.1868-1876
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• 2010

In this paper, we design a 256-bit EEPROM IP using only logic process-based devices. We propose EEPROM core circuits, a control gate (CG) and a tunnel gate (TG) driving circuit, to limit the voltages between the devices within 5.5V; and we propose DC-DC converters : VPP (=+4.75V), VNN (-4.75V), and VNNL (=VNN/3) generation circuit. In addition, we propose switching powers, CG_HV, CG_LV, TG_HV, TG_LV, VNNL_CG, VNNL_TG switching circuit, to be supplied for the CG and TG driving circuit. Simulation results under the typical simulation condition show that the power consumptions in the read, erase, and program mode are $12.86{\mu}W$, $22.52{\mu}W$, and $22.58{\mu}W$ respectively. Furthermore, the manufactured test chip operated normally and generated its target voltages of VPP, VNN, and VNNL as 4.69V, -4.74V, and -1.89V.

• ETRI Journal
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• v.37 no.6
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• pp.1188-1198
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• 2015

In this paper, a multi-time programmable (MTP) cell based on a $0.18{\mu}m$ bipolar-CMOS-DMOS backbone process that can be written into by using dual pumping voltages - VPP (boosted voltage) and VNN (negative voltage) - is used to design MTP memories without high voltage devices. The used MTP cell consists of a control gate (CG) capacitor, a TG_SENSE transistor, and a select transistor. To reduce the MTP cell size, the tunnel gate (TG) oxide and sense transistor are merged into a single TG_SENSE transistor; only two p-wells are used - one for the TG_SENSE and sense transistors and the other for the CG capacitor; moreover, only one deep n-well is used for the 256-bit MTP cell array. In addition, a three-stage voltage level translator, a VNN charge pump, and a VNN precharge circuit are newly proposed to secure the reliability of 5 V devices. Also, a dual memory structure, which is separated into a designer memory area of $1row{\times}64columns$ and a user memory area of $3rows{\times}64columns$, is newly proposed in this paper.

• Journal of fish pathology
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• v.34 no.2
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• pp.133-140
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• 2021

With the recent isolation of a new betanodavirus in shellfish, Korean Shellfish Nervous Necrosis Virus (KSNNV), it has also been identified the reassortant KSNNV of two RNA segments, in which one segment is KSNNV genotype but the other one is known genotype. In this study, we confirmed that the ressortant KSNNVs obtained in previous screening study of our laboratory for betanodaviruses in shellfish were KS/RGNNV and RG/KSNNV type by performing two consecutive multiplex RT-PCR on each RNA1 and RNA2 segment (R1- and R2-discriminative multiplex two-step RT-PCR, respectively) to determine the genotype of each segment based on the size of amplicon. In the pathogenicity analysis, none of the reassortants induced specific external symptoms or mortality of VNN, but viruses of 2 × 10⁴~10⁵ copies/mg or more were detected at 14 days after injection (10⁷ copies/fish) in brain tissues of 4 species except for crucian carp and common carp among the 6 species of juvenile fish used. In addition, the histopathological features of weak but distinct vacuole formation were also found in the brain of these infected fish, but no difference was found between the two reassortants KS/RGNNV-KG and RG/KSNNV-CM.

• Proceedings of the Korean Institute of Information and Commucation Sciences Conference
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• 2012.10a
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• pp.852-855
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• 2012

A DC-DC converter for EEPROM IPs which perfom erasing by the FN (Fowler-Nordheim) tunneling and programming by the band-to-band tunneling is designed in this paper. For the DC-DC converter for EEPROM IPs using a low voltage of $1.5V{\pm}10%$ as the logic voltage, a scheme of using VRD (Read Voltage) instead of VDD is proposed to reduce the pumping stages and pumping capacitances of its charge pump circuit. VRD ($=3.1V{\pm}0.1V$) is a regulated voltage by a voltage regulator using an external voltage of 5V. The designed DC-DC converter outputs VPP (=8V) and VNN (=-8V) in the write mode.

• Journal of IKEEE
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• v.28 no.1
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• pp.78-89
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• 2024

PMIC chips based on a BCD process used in automotive semiconductors require multi-time programmable (MTP) intellectual property (IP) that does not require additional masks to trim analog circuits. In this paper, MTP cell size was reduced by about 18.4% by using MTP cells using PMOS capacitors (PCAPs) instead of NMOS capacitors (NCAPs) in MTP cells, which are single poly EEPROM cells with two transistors and one MOS capacitor for small-area MTP IP design. In addition, from the perspective of MTP IP circuit design, the two-stage voltage shifter circuit is applied to the CG drive circuit and TG drive circuit of MTP IP design, and in order to reduce the area of the DC-DC converter circuit, the VPP (=7.75V), VNN (=-7.75V) and VNNL (=-2.5V) charge pump circuits using the charge pumping method are placed separately for each charge pump.