• Reproductive and Developmental Biology
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• v.33 no.4
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• pp.243-248
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• 2009

To examine the differential expression of proteins during the cycling (70~80% confluences) and G0/G1 (full confluences) phases in porcine fetal fibroblast cells, we used a global proteomics approach by 2-D gel electrophoresis (2-DE) and MALDI-TOF-MS. Cycling cell were harvested at approximately 70% to 80% confluent state while cells in G0/G1 phase were recovered after maintenance of a confluent state for 48 hr. Cellular proteins with isoelectric points ranging between 3.0~10.0, were analyzed by 2-DE with 2 replicates of each sample. A total of approximately 700 spots were detected by 2.D gels stained with Coomassie brilliant blue. On comparing the cell samples obtained from the cycling and G0/G1 phases, a total of 13 spots were identified as differentially expressed proteins, of which 8 spots were up-regulated in the cycling cell and 5 were up-regulated in the G0/G1 phase. Differentially expressed proteins included K3 keratin, similar to serine protease 23 precursor, protein disulfide-isomerase A3, microsomal protease ER-60, alpha-actinin-2, and heat-shock protein 90 beta. The identified proteins were grouped on the basis of their basic functions such as molecular binding, catabolic, cell growth, and transcription regulatory proteins. Our results show expression profiles of key proteins in porcine fetal fibroblast cells during different cell cycle status.

• The Journal of Advanced Prosthodontics
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• v.6 no.1
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• pp.8-13
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• 2014

PURPOSE. The purpose of this study was to evaluate whether surface treatments affect the translucency of laminate veneers with different shades and thicknesses. MATERIALS AND METHODS. A total of 224 disc-shaped ceramic veneers were prepared from A1, A3, HT (High Translucent) and HO (High Opaque) shades of IPS e.max Press (Ivoclar Vivadent) with 0.5 mm and 1.0 mm thicknesses. The ceramics were divided into four groups for surface treatments. Group C: no surface treatments; Group HF: etched with hydrofluoric acid; Group SB: sandblasted with 50-${\mu}m$ $Al_2O_3$; and Group L; irradiated with an Er;YAG laser. A translucent shade of resin cement (Rely X Veneer, 3M ESPE) was chosen for cementation. The color values of the veneers were measured with a colorimeter and translucency parameter (TP) values were calculated. A three-way ANOVA with interactions for TP values was performed and Bonferroni tests were used when appropriate (${\alpha}=0.05$). RESULTS. There were significant interactions between the surface treatments, ceramic shades and thicknesses (P=.001). For the 0.5-mm-thick specimens there were significant differences after the SB and L treatments. There was no significant difference between the HF and C treatments for any shades or thicknesses (P>.05). For the 1-mm-thick ceramics, there was only a significant difference between the L and C treatments for the HT shade ceramics (P=.01). There were also significant differences between the SB and C treatments except not for the HO shades (P=.768). CONCLUSION. The SB and L treatments caused laminate veneers to become more opaque; however, HF treatment did not affect the TP values. When the laminate veneers were thinner, both the shade of the ceramic and the SB and laser treatments had a greater effect on the TP values.

• Journal of Microbiology and Biotechnology
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• v.31 no.1
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• pp.16-24
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• 2021

Hepatitis B virus (HBV) genome P-encoded protein HBV DNA polymerase (Pol) has long been known as a reverse transcriptase during HBV replication. In this study, we investigated the impact of HBV Pol on host cellular processes, mainly apoptosis, and the underlying mechanisms. We showed a marked reduction in apoptotic rates in the HBV Pol-expressed HepG2 cells compared to controls. Moreover, a series of assays, i.e., yeast two-hybrid, GST pull-down, co-immunoprecipitation, and confocal laser scanning microscopy, identified the host factor eEF1A2 to be associated with HBV Pol. Furthermore, knockdown of eEF1A2 gene by siRNA abrogated the HBV Pol-mediated anti-apoptotic effect with apoptosis induced by endoplasmatic reticulum (ER) stress-inducer thapsigargin (TG), thus suggesting that the host factor eEF1A2 is essential for HBV Pol's anti-apoptosis properties. Our findings have revealed a novel role for HBV Pol in its modulation of apoptosis through integrating with eEF1A2.

• Journal of Korean Neurosurgical Society
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• v.65 no.1
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• pp.4-12
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• 2022

Objective : We reported the differentially methylated genes in patients with subarachnoid hemorrhage (SAH) using bioinformatics analyses to explore the biological characteristics of the development of delayed cerebral ischemia (DCI). Methods : DNA methylation profiles obtained from 40 SAH patients from an epigenome-wide association study were analyzed. Functional enrichment analysis, protein-protein interaction (PPI) network, and module analyses were carried out. Results : A total of 13 patients (32.5%) experienced DCI during the follow-up. In total, we categorized the genes into the two groups of hypermethylation (n=910) and hypomethylation (n=870). The hypermethylated genes referred to biological processes of organic cyclic compound biosynthesis, nucleobase-containing compound biosynthesis, heterocycle biosynthesis, aromatic compound biosynthesis and cellular nitrogen compound biosynthesis. The hypomethylated genes referred to biological processes of carbohydrate metabolism, the regulation of cell size, and the detection of a stimulus, and molecular functions of amylase activity, and hydrolase activity. Based on PPI network and module analysis, three hypermethylation modules were mainly associated with antigen-processing, Golgi-to-ER retrograde transport, and G alpha (i) signaling events, and two hypomethylation modules were associated with post-translational protein phosphorylation and the regulation of natural killer cell chemotaxis. VHL, KIF3A, KIFAP3, RACGAP1, and OPRM1 were identified as hub genes for hypermethylation, and ALB and IL5 as hub genes for hypomethylation. Conclusion : This study provided novel insights into DCI pathogenesis following SAH. Differently methylated hub genes can be useful biomarkers for the accurate DCI diagnosis.

• Journal of Animal Science and Technology
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• v.49 no.3
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• pp.309-320
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• 2007

The purpose of the present study was to determine if monocarboxylate transporter(MCT) isoforms and Basigin(Bsg) are expressed in the efferent ductules(ED) and if MCT1 expression is under estrogen receptor(ER)α-regulation in the ED of male reproductive tract. The presence of MCT isoforms and Bsg mRNAs was detected by real-time polymerization chain reaction(PCR), and ERα-mediated regulation of MCT1 expression in the ED was indirectly determined by immuno- histochemistry. Current study found differential expression of MCT isoforms(MCT1, 2, 3, 4, and 8) and Bsg mRNAs in rat ED according to postnatal ages. In addition, comparison of MCT1 expression in the ED between wild type and ERα knockout mice at different postnatal ages showed basolateral localization of MCT1 in ciliated cells of the ED and, in part, ERα- mediated regulation of MCT1 expression. It is suggested that MCTs would play a role in regulation of function of the ED.

• Journal of Plant Biotechnology
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• v.1 no.1
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• pp.20-30
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• 1999

In order to modify lipid production of Perilla qualitatively as well as quantitatively by genetic engineering, genes involved in carbon metabolism were isolated and characterized. These include acyl-ACP thioesterases from Perilla frutescens and Iris sp., four different $\beta$-ketoacyl- ACP synthases from Perilla frutescens, and two $\Delta$15 a-cyl-ACP desaturases(Pffad7, pffad3). Δ15 acyl-ACP desa turase (Δ15-DES) is responsible for the conversion of linoleic acid (18:2) to $\alpha$-linolenic acid (ALA, 18:3). pffad 3 encodes Δ15 acyl-desaturase which is localized in ER membrane. On the other hand, Pffad7 encodes a 50 kD plastid protein (438 residues), which showed highest sequence similarity to Sesamum indicum fad7 protein. Northern blot analysis revealed that the Pffad7 is highly expressed in leaves but not in roots and seeds. And Pffad3 is expressed throughout the seed developmental stage except very early and fully mature stage. We constructed Pffad7 gene under 355 promoter and Pffad3 gene under seed specific vicillin promoter. Using Pffad7 construct, Perilla, an oil seed crop in Korea, was transformed by Agrobacterium leaf disc method. $\alpha$-linolenic acid contents increased in leaves but decreased in seeds of transgenic Perilla. Currently, we are transforming Perilla with Pffad3 construct to change Perilla seed oil composition. We isolated three ADP-glucose pyrophosphorylase (AGP) genes from Perilla immature seed specific cDNA library. Nucleotide sequence analysis showed that two of three AGP (Psagpl, Psagp2) genes encode AGP small subunit polypeptides and the remaining (Plagp) encodes an AGP large subunit. PSAGPs, AGP small subunit peptide, form active heterotetramers with potato AGP large subunit in E. coli expressing plant AGP genes.

• Journal of Korean Philosophical Society
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• v.129
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• pp.315-339
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• 2014

Diese Abhandlung untersucht Jacobis ${\ddot{U}}ber$ die Lehre Spinoza und den von diesem veranlassten Spinoza-Streit. Damit sie $enth{\ddot{u}}llt$ zuerst Jacobischen Zusammenhang zwischen transzent und immanent und folgt auf seine Wirkungsgeschichte in der Moderne. Ich rekonstruiere den Streit zwischen Jacobi und Lessing und danach interpretiere dessen Rezeption durch Hegel und Schleiermacher. Lessing stellt anstatt der traditionellen Begriffe der Gottheit ἑν ${\kappa}{\alpha}{\iota}$ ${\pi}{\alpha}{\nu}$ auf. $Demgegen{\ddot{u}}ber$ behauptet Jacobi Salto mortale um ihn ${\ddot{u}}berschreiten$ zu $k{\ddot{o}}nnen$, indem er Lessing als Pantheist und Atheist bestimmt. Salto mortale bei Jacobi ist der Sprung zu dem ${\ddot{U}}bernat{\ddot{u}}rlichen$ und dem Glaube. Der Streit zwischen Jacobi und Lessing ist der zwischen dem Naturalismus und ${\ddot{U}}bernaturalismus$ und $dar{\ddot{u}}berhinaus$ der zwischen dem Athismus und Theismus. $W{\ddot{a}}hrend$ die Natur der Inbegriff der Bedingten ist, ist Gott der absolute Anfang der Natur $au{\ss}erhalb$ des Naturzusammenhangs. $W{\ddot{a}}hrend$ Spinoza Gott im $nat{\ddot{u}}rlichen$ Zusammenhang begreift, $fa{\ss}t$ Jacobi den im ${\ddot{u}}bernat{\ddot{u}}rlichen$ auf. Deus sive natura bei Spinoza $ver{\ddot{a}}ndert$ sich Gott im Menschen bei Jacobi. Gott im Menschen ist nichts anders als das Prinzip des Lebens und das aller Vernuft. In diesem Zusammenhang $fa{\ss}t$ Hegel Gott als Geist denn Subjekt des Lebens auf und $h{\ddot{a}}lt$ das Wesen des Geistes $f{\ddot{u}}r$ die sich selbst vermittelnde Bewegung. Dies zeigt sich als die Spinoza ${\ddot{u}}berbietende$ Immanenzphilosophie. $Demgegen{\ddot{u}}ber$ behauptet Schleiermacher die Einheit des Endlichen und Unendlichen in der $religi{\ddot{o}}sen$ Anschuung. Die Verbindung von Mensch und Gott ist die im Endlichen immanent bleibende Anschauung der $g{\ddot{o}}ttlichen$ Eigenschaft. Dies zeigt das transzendente im immanenten.

• Korean Journal of Poultry Science
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• v.44 no.1
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• pp.1-9
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• 2017

The objective of the present study was to determine the expression of genes associated with lipopolysaccharide (LPS)-induced stressor in two breeds of chickens: the Korean native chicken (KNC) and the White Leghorn chicken (WLH). Forty chickens per breed, aged 40 weeks, were randomly allotted to the control (CON, administered the saline vehicle) and LPS-injected stress groups. Samples were collected at 0 and 48 h post-LPS injection, and total RNA was extracted from the chicken livers for RNA microarray and quantitative real-time polymerase chain reaction (qRT-PCR) analyses. In response to LPS, 1,044 and 1,193 genes were upregulated, and 1,000 and 1,072 genes were downregulated in the KNC and WLH, respectively, using a ${\geq}2$-fold cutoff change. A functional network analysis revealed that stress-related genes were downregulated in both KNC and WLH after LPS infection. The results obtained from the qRT-PCR analysis of mRNA expression of heat shock 90 (HSP90), 3-hydroxy-3-methylglutaryl-CoA reductase (HMGCR), activating transcription factor 4 (ATF4), sterol regulatory element-binding protein 1 (SREBP1), and X-box binding protein 1 (XBP1) were confirmed by the results of the microarray analysis. There was a significant difference in the expression of stress-associated genes between the control and LPS-injected KNC and WLH groups. The qRT-PCR analysis revealed that the stress-related $HSP90{\alpha}$ and HMGCR genes were downregulated in both LPS-injected KNC and WLH groups. However, the HSP70 and $HSP90{\beta}$ genes were upregulated only in the LPS-injected KNC group. The results suggest that the mRNA expression of stress-related genes is differentially affected by LPS stimulation, and some of the responses varied with the chicken breed. A better understanding of the LPS-induced infective stressors in chicken using the qRT-PCR and RNA microarray analyses may contribute to improving animal welfare and husbandry practices.

• Development and Reproduction
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• v.17 no.4
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• pp.441-449
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• 2013

Recent study showed that T cells in the immune organs and peripheral blood are influenced by estradiol, leading to a dysfunction of the immune system. However, little is known about the thymic-gonadal relationship during the estrous cycle in mouse. Therefore, the purpose of this study was to elucidate the mechanism by which a change in estradiol levels during the estrous cycle regulates the development of T cells in the mouse thymus. Six-week-old ICR mice were used and divided into four groups, including diestrous, proestrous, estrous, and metestrous. We first confirmed that ER-${\alpha}$ and - ${\beta}$ estrogen receptors were expressed in thymic epithelial cells, showing that their expression was not different during the estrous cycle. There was also no significant difference in thymic weight and total number of thymocytes during the estrous cycle. To determine the degree of thymocyte differentiation during the estrous cycle, we analyzed thymocytes by flow cytometry. As a result, the percentage of CD4+CD8+ double-positive (DP) T cells was significantly decreased in the proestrous phase compared to the diestrous phase. However, CD4+CD8- or CD4-CD8+ (SP) T cells were significantly increased in the proestrous phase compared to the diestrous phase. In addition, the percentage of CD44+CD25- (DN1) T cells was significantly decreased in the estrous phase compared to other phases, whereas the percentages of CD44+CD25+ (DN2), CD44-CD25+ (DN3), and CD44-CD25- (DN4) were not changed during the estrous cycle. These results indicate that the development of thymocytes may arrest in the DP to SP transition stage in the proestrous phase displaying the highest serum level of estradiol. This study suggests that a change in estradiol levels during the estrous cycle may be involved in the regulation of thymocyte differentiation in the mouse thymus.

• Asian Pacific Journal of Cancer Prevention
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• v.13 no.11
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• pp.5339-5343
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• 2012

The induction of apoptosis in target cells is a key mechanism for most anti-tumor therapies. Bufalin is a cardiotonic steroid that has the potential to induce differentiation and apoptosis of tumor cells. Research on bufalin has so far mainly involved leukemia, prostate cancer, gastric cancer and liver cancer, and has been confined to in vitro studies. The bufadienolides bufalin and cinobufagin have been shown to induce apoptosis in a wide spectrum of cancer cell. The present article reviews the anticancer effects of bufalin. It induces apoptosis of lung cancer cells via the PI3K/Akt pathway and also suppressed the proliferation of human non-small cell lung cancer A549 cell line in a time and dose dependent manner. Bufalin, bufotalin and gamabufotalin, key bufadienolides, significantly sensitize human breast cancer cells with differing ER-alpha status to apoptosis induction by the TNF-related apoptosis-inducing ligand (TRAIL). In addition, bufadienolides induce prostate cancer cell apoptosis more significantly than that in breast epithelial cell lines. Similar effects have been observed with hepatocellular carcinoma (HCC) but the detailed molecular mechanisms of inducing apoptosis in this case are still unclear. Bufalin exerts profound effects on leukemia therapy in vitro. Results of multiple studies indicate that bufalin has marked anti-tumor activities through its ability to induce apoptosis. Large-scale randomized, double-blind, placebo or positive drug parallel controlled studies are now required to confirm the efficacy and apoptosis-inducing potential of bufalin in various cancers in the cliniucal setting.