• 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.

• Proceedings of the Korean Institute of Information and Commucation Sciences Conference
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• 2015.05a
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• pp.832-836
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• 2015

In this paper, a DC-DC converter is designed for logic process MTP (multi-time programmable) memory IPs using dual program voltage, which are used for analog trimming or storing chip IDs in sensor applications. The DC-DC converter supplies VPP (=5.25V), VNN (=-5.25V), and VNNL ($=2{\cdot}VNN/5$). It uses MOS capacitors and designed with only 3,3V devices. VPP and VNN are configured in two and five stages, respectively. And their pumping currents are $9.17{\mu}A$ and $9.7{\mu}A$, respectively.

• Journal of fish pathology
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• v.25 no.2
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• pp.111-116
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• 2012

Prevalence of viral nervous necrosis (VNN) in sevenband grouper Epinephelus septemfasciatus farms was investigated during the period of 2006-2008. The outbreak of the VNN was observed from August (water temperature: $24-26^{\circ}C$) to September or October ($20-25^{\circ}C$). The viral infection resulted in acute or chronic mortality of the host, where the mortality rate of juvenile fish was higher than that of the adult fish. Phylogenetic analysis based on partial RNA2 coat protein gene nucleotide sequences revealed that all the isolates from sevenband grouper were classified into the genotype redspotted grouper nervous necrosis virus (RGNNV). In conclusion, VNN of juvenile and adult sevenband groupers during the summer season (July to October, water temperature: about $24^{\circ}C$) was caused by virus belonging to the genotype RGNNV.

• 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.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.

• 한국전산유체공학회:학술대회논문집
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• 2009.04a
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• pp.269-276
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• 2009

The local preconditioning method has both robust convergence and accurate solutions by using local flow properties for parameters in the preconditioning matrix. Preconditioning methods have been very effective to low speed inviscid flows. In the viscous and turbulent flows, deterioration of convergence should be overcame on the high aspect ratio grids to get better convergence and accuracy. In the present study, the local time stepping and min-CFL/max-VNN definitions are applied to compare the results and we propose the method that switches between two methods. The min-CFL definition is applied for inviscid flow problems and the min-CFL/max-VNN definition is implemented to viscous and turbulent flow problems.

• 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.

• 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.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.35 no.3
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• pp.237-241
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• 2002

This study was performed both to explore the host of nervous necrosis virus (NNV) between mariculturing fish species and to examine the phylogenic position of the NNV in Korea. NNV was confirmed on the basis of histopathological and molecular biological examination, then VNN infection was Preyed from either moribund or dead fishes including red drum, Sciaenops ocellatus; oblong rock fish, Sebastes oblongus and flounder, Paralichthys olivaceus. As a result of sequencing for a part of ms, virus from red drum was showed $98\%, $97\%, $86\% and $74\% homology with oblong rock fish, grouper, Japanese flounder and striped jack, respectively. On the other hand, NNV from oblong rock fish was demonstrated $96\%, $85\% and $72\% homology with grouper, Japanese flounder and striped jack, respectively. NNV from red drum and oblong rock fish was exhibited phylogenically distant from the representative NNV, SJNNV originated from striped jack. On the contrary, the viruses appeared to be similar species with Taiwan NNV isolated from culturing grouper.