• Proceedings of the PSK Conference
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• 2003.04a
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• pp.185.1-185.1
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• 2003

In this study, we investigated the effect of cadmium on genes expression related to zinc homeostasis in HT22 hippocampal neuron cell line by RT -PCR and western blotting technics. In the time-course effect, cadmium up-regulated the relative levels of MT -I and MT -II to~b-actin at 4 hr after treatment. These effects were consistent with MT -I/II protein contents by western blot analysis. But MT -III, a specific MT isoform in brain, was not affected by cadmium. (omitted)

• Molecules and Cells
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• v.20 no.1
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• pp.51-56
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• 2005

Excitation-contraction coupling (ECC) proteins in the human heart were characterized using human atrial tissues from different age groups. The samples were classified into one infant group (Group A: 0.2-7 years old) and three adult groups (Group B: 21-30; Group C: 41-49; Group D: 60-66). Whole homogenates (WH) of atrial tissues were assayed for ligand binding, $^{45}Ca^{2+}$ uptake and content of ECC proteins by Western blotting. Equilibrium [$^3H$]ryanodine binding to characterize the ryanodine receptor (RyR) of the sarcoplasmic reticulum (SR) showed that the maximal [$^3H$]ryanodine binding ($B_{max}$) to RyR was similar in all the age groups, but the dissociation constant ($k_d$) of ryanodine was higher in the infant group than the adult groups. Oxalate-supported $^{45}Ca^{2+}$ uptake into the SR, a function of the SR SERCA2a activity, was lower in the infant group than in the adult groups. Similarly, [$^3H$]PN200-110 binding, an index of dihydropyridine receptor (DHPR) density, was lower in the infant group. Expression of calsequestrin and triadin assessed by Western blotting was similar in the infant and adult groups, but junctin expression was considerably higher in the adult groups. These differences in key ECC proteins could underlie the different $Ca^{2+}$ handling properties and contractility of infant hearts.

• Asian Pacific Journal of Cancer Prevention
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• v.13 no.11
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• pp.5709-5714
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• 2012

Objective: To evaluate the effects of curcumin on matrixmetalloproteinase-9 (MMP-9) and invasion ability induced by transforming growth factor-${\beta}1$ (TGF-${\beta}1$) in MDA-MB-231 cells and potential mechanisms. Methods: Human breast cancer MDA-MB-231 cells were used with the CCK-8 assay to measure the cytotoxicity of curcumin. After treatment with 10 ng/ml TGF-${\beta}1$, with or without curcumin (${\leq}10{\mu}M$), cell invasion was checked by transwell chamber. The effects of curcumin on TGF-${\beta}1$-stimulated MMP-9 and phosphorylation of Smad2, extracellular-regulated kinase (ERK), and p38 mitogen activated protein kinases (p38MAPK) were examined by Western blotting. Supernatant liquid were collected to analyze the activity of MMP-9 via zymography. Following treatment with PD98059, a specific inhibitor of ERK, and SB203580, a specific inhibitor of p38MAPK, Western blotting and zymography were employed to examine MMP-9 expression and activity, respectively. Results: Low dose curcumin (${\leq}10{\mu}M$) did not show any obvious toxicity to the cells, while $0{\sim}10{\mu}mol/L$ caused a concentration-dependent reduction in cell invasion provoked by TGF-${\beta}1$. Curcumin also markedly inhibited TGF-${\beta}1$-regulated MMP-9 and activation of Smad2, ERK1/2 and p38 in a dose- and time-dependent manner. Additionally, PD98059, but not SB203580, showed a similar pattern of inhibition of MMP-9 expression. Conclusion: Curcumin inhibited TGF-${\beta}1$-stimulated MMP-9 and the invasive phenotype in MDA-MB-231 cells, possibly associated with TGF-${\beta}$/Smad and TGF-${\beta}$/ERK signaling.

• Fisheries and Aquatic Sciences
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• v.23 no.3
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• pp.8.1-8.9
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• 2020

Background: The present study investigates the potent skin whitening ability of ethanol extract from the brown alga, Padina boryana (PBE) which was collected in the shores of Fulhadhoo Island, the Maldives, and its specific pathways of action. The effect of PBE which contains a rich amount of polyphenols was evaluated using B16F10 murine melanoma cells and provides insight to the underlying mechanisms with reference to the inhibition of melanin formation. Methods: Melanin synthesis and cellular tyrosinase inhibition were assessed in the α-MSH-stimulated melanocytes. Melanogenic pathway-related protein expressions were investigated via Western blotting. ERK 42/44 was particularly examined considering its involvement in the melanogenic pathway. Further, RT-qPCR techniques were involved in gene expression analysis. Results: PBE dose-dependently inhibited the cellular melanin synthesis and tyrosinase levels. Western blotting revealed the potential of PBE to downregulate microphthalmia-associated transcription factor (MITF), tyrosinase, and tyrosinase-related protein-1 and protein-2 (TRP-1 and TRP-2). Moreover, results explained the phosphorylation of ERK was sustained via PBE and hence declined the ultimate melanin synthesis. Gene expression analysis reinforced the results obtained. Conclusions: The study provides substantial evidence to express the potential of PBE to inhibit B16F10 melanoma cell melanin synthesis. Concisely, results suggest the ability of PBE to be involved in medicinal and cosmeceutical applications.

• Journal of Environmental Science International
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• v.12 no.6
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• pp.651-657
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• 2003

In order to examine if arsenic, one of environmental stresses, contributes to hypertension as one of cardiovas cular pathological factors, this study was perfarmed in vivo and in vitro, using intacted or pithed rats and aorta ring preparation, respectively. And also the relationship between expression of heat shock protein (HSP) 90 and vasoactives-induced contractile response was elucidated. To measure blood pressure, the carotid arterial pressure was recorded on physiograph(Grass Co. 79E) connected to strain gauge. On the other hand, contractile response of vascular ring preparation isolated from rat was determined in organ bath and was recorded on physiograph connected to isometric transducer. And HSP was detacted by Western blotting whole cell Iysis. Preganglionic nerve stimulation was increased by 26.0% in arterial pressure of rat treated with arsenic. Vascular contractile response was monitored and HSP were measured by Western blotting of whole Iysis prepared from samples exposed with 0, 0.5, 1, 2 and 4 mM of arsenic for 8 hours. The dose-vascular responses of potassium chloride were augmented by increasing dose of arsenic in the strips exposed to arsenic for 8 hours, and were not augmented for 1, 3, 5 hours. And the response of relaxation of rat aorta induced by histamine was not influenced by arsenic stress. The increase of HSP 90 expression in rat aorta was pronounced at 8 hours after 4 mM of arsenic treatment, but HSP 60 expression was not. Arsenic stress not only increased the expression of HSP 90 in the rat aorta, but also augmented contractions to potassium chloride. These results indicated that arsenic stress was sufficient to induce heat shock protein 90, resulting in increased vascular contractility in rat aorta.

• Journal of Life Science
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• v.24 no.5
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• pp.485-490
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• 2014

Poria cocas has been reported to be effective in skin whitening. However, the direct effect of P. cocas ethanol extracts (PCEE) on melanin synthesis has not been scientifically studied. To elucidate the direct effect of PCEE on melanogenesis, a 3,4-dihydroxyindole-2-carboxylic acid (DOPA) synthesis assay, tyrosinase activity assay, and Western blotting for melanogenic proteins, including tyrosinase, tyrosinase-related protein (TRP)-1, and TRP-2 were performed in mouse B16F1 cells. The results revealed that PGEE inhibited melanin production in a dose-dependent manner by blocking the synthesis of DOPA. Although the activation of tyrosinase was not affected, the expression levels of TRP-1 and TRP-2 were controlled. These results suggest that PCEE has a whitening effect, indicating that it may be a useful agent in the development of whitening cosmetics.

• Journal of Genetic Medicine
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• v.4 no.2
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• pp.179-185
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• 2007

Purpose : Catecholamines are the neuro-transmitters in the sympathetic nervous system (SNS) and are activated by stress stimulus. Tyrosine hydroxylase (TH) and Dopamine-${\beta}$-Hydroxylase (DBH) are very important enzymes in the catecholamine synthesis. Corticotropin releasing hormone (CRH) is released in the process of reacting to stresses. The aim of this study is to find out what effects immobilization stresses have on the expression of TH, BDH and CRH mRNA in a rat's brains. Methods : We compare expression levels in rat's brains of TH, DBH and CRH mRNA induced by immobilization stresses between the test group and controled group. The expression levels of TH, DBH and CRH mRNA are measured by RT-PCR and the Western Blotting Analysis (WBA). Results : In brains and adrenal glands of the immobilization stress group, the expression levels of TH and DBH mRNAs are significantly two to three times higher (P<0.01), and CRH mRNAs are approximately one and a half times higher (P<0.05) than those of controlled group. Conclusion : This study suggest that the expression levels of TH, DBH and CRH mRNAs are activated by stress stimulus in a rat's brains and adrenal glands.

• Korean Journal of Animal Reproduction
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• v.27 no.1
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• pp.53-59
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• 2003

For somatic cell nuclear transfer in Hanwoo, fetal fibroblast cell lines were established from 35, 50, 70 and 90-day fetuses of Korean native cattle. The sex of these fetal fibroblast cells were analyzed by PCR using Y-specific primers and confirmed that two cell lines were female and the other two cell lines were male. Karyotyping of these cell lines indicates that the chromosome numbers of fetal fibroblast cells were not affected by passage number and more than 80% of fetal fibroblast cells have normal chromosome number. To evaluate Go stage in cell cycle of fetal fibroblast cells, Western blotting was performed to detect the expression level of PCNA which is known to be expressed in all cell cycle stages except G$_{0}$ stage. Following serum starvation or confluent culture for 7 days, fetal fibroblast cells were effectively reached to G$_{0}$ stage. The cell cycle was resumed after culture of these Go stage-fetal fibroblast cells with normal medium. These results indicates that fetal fibroblast cells originated from Hanwoo were successfully isolated and culture system and induction of cell cycle of these cells were established for somatic cell nuclear transfer in Hanwoo.woo.

• Parasites, Hosts and Diseases
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• v.45 no.1 s.141
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• pp.19-26
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• 2007

This study describes the isolation of a Toxocara canis species-specific excretory-secretory(ES) antigen and the development of an enzyme-linked immunosorbent assay(ELISA) based on this antigen. Analysis of the ES antigens of T. canis, Toxocara vitulorum, Ascaris lumbricoides and Necator americanus larval antigen was performed by SDS-PAGE followed by western blotting. A 57 kDa T. canis-specific antibody fraction(TcES-57) was identified by western blotting and labelling with anti-Toxocara antibodies(from experimental rabbits and human patients) and tracing with anti-human or anti-rabbit peroxidase conjugate. No protein fraction of 57 kDa was detected in ES or larval antigens collected from T. canis, T. vitulorum, A. lumbricoides and N. americanus. Using TcES-57, a specific anti-serum was produced in rabbits and a double sandwich ELISA was developed. This test was validated using known seropositive sera from toxocariasis patients, sera from A. lumbricoides or N. americanus patients, and 50 serum samples from cats. These tests revealed that TcES-57 antigen is specific to T. canis infection and does not cross react with sera of other related infections. Thus, ELISA based on TcES-57 antigen was proven to be an effective tool in the diagnosis of toxocariasis and studies on the role of T. canis in the epidemiology of human toxocariasis.

• Proceedings of the KSAR Conference
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• 2003.06a
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• pp.97-97
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• 2003

For many animals the centrosome consists of a pair of centrioles and surrounding pericentriolar materials (PCMs). PCMs have been known to play roles during cell division. It is known that centrioles are necessary to assemble centrosomal components. However, many types of oocytes undergo meiosis without centrioles. It is known that in nonmurine mammalian species, the sperm introduces an intact proximal centriole unlike sea urchin where two centrioles are introduced. In case of mouse sperm, the presence of centrosome is not clear In this study, a monoclonal antibody was developed to investigate centrosome during mouse germ cell and early embryo development. Results of immunostaining and Western blotting in CHO cells suggest that the monoclonal antibody recognizes a nuclear and centrosomal protein, thus called NCP. The NCP monoclonal antibody was used to screen a cDNA expression library prepared from 12.5 mouse brain to isolate NCP gene. Nucleotide size of NCP gene obtained from immunoscreening was about 5.5kb. It is determined that the NCP may be closely related with pericentriolar material -1 gene (Pcm-1) from the result of sequencing analysis. The molecular weight, 66kDa, calculated by known DNA sequence in database is consistent with that of detected from Western blotting using CHO cell lysates. Therefore, it is assumed that NCP may be alternative splicing form of Pcm-1 of which molecular weight is 228kDa. In mouse oocytes, NCP was distributed in nucleus as in CHO cells. It was shown that the NCP was localized around neck region, probably the centrosome in mouse neck region. Interestingly, dramatic change in distribution of NCP was also shown in male germ cell development. Finally, we observed the cellular distribution of NCP during early embryo development. NCP was detected in nucleus as well as centrosome foci. It is suggested that the centrioles reassembly we occurring in blastocysts and then affects the distribution of NCP.