• Development and Reproduction
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• v.16 no.1
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• pp.39-45
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• 2012

To verify the sex steroids which are involved in oocyte maturation of the blacktip grouper, $Epinephelus$ $fasciatus$, we incubated vitellogenic oocytes (0.41 and 0.50 mm in average diameter) in the presence of exogenous steroid precursor ($[^3H]17{\alpha}$-hydroxyprogesterone). Steroids were extracted, separated and identified by thin layer chromatography. The major metabolites produced were androstenedione, estradiol-$17{\beta}$, estrone and progestogens. Progestogen metabolites in the oocytes of 0.50 mm were more abundant than those of 0.41 mm. Also, we investigated the $in$ $vitro$ effects of human chorionic gonadotropin (HCG; 5, 50 and 500 $IU/m{\ell}$), $17{\alpha},20{\beta}$-dihydroxy-4-pregnen-3-one ($17{\alpha}20{\beta}P$) and $17{\alpha},20{\beta}$-trihydroxy-4-pregnen-3-one ($17{\alpha}20{\beta}21P$; 5, 50 and 500 $ng/m{\ell}$, respectively) on oocyte maturation. In the oocytes of 0.41 mm, treatment with 50 IU HCG stimulated GVBD ($55.30{\pm}1.20%$) compared with controls ($32.41{\pm}3.13%$, $p$<0.05). In the oocytes of 0.50 mm, treatment of $17{\alpha}20{\beta}P$ (50 and 500 $ng/m{\ell}$) stimulated GVBD ($50.13{\pm}2.52$ and $51.77{\pm}5.91%$, respectively) compared with controls ($36.81{\pm}2.89%$, $p$<0.05). Treatment with 500 IU HCG also stimulated GVBD ($49.59{\pm}5.15%$) compared with controls ($p$<0.05). Taken together, these results suggested that both HCG and $17{\alpha}20{\beta}P$ were effective on in vitro oocyte maturation and $17{\alpha}20{\beta}P$ may act as a maturation inducing hormone in blacktip grouper.

• Journal of Food Hygiene and Safety
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• v.33 no.5
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• pp.339-346
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• 2018

This study was conducted to monitor the current status of pesticide residues in a total of 297 agricultural products for raw juice from January 2017 to September 2017. Pesticide residues in samples were analyzed by multiresidue method for the determination of multiclass pesticide with an aim to detect 220 pesticides using GC/ECD, GC/NPD, TOF/MS, LC/PDA, LC/FLD, and LC/MS-MS. The analysis revealed the presence of nine pesticides in 14 samples. The detected pesticides were chlorpyrifos, cyflufenamid, chlorothalonil, pendimethalin, azoxystrobin, pyridalyl, imazalil, fludioxonil and procymidone. The kinds of detected crops were mandarin, carrot, apple, celery, chicory, orange and grape. Risk assessment evaluated human health exposure with the ratio of EDI (Estimated daily intake) to ADI (Acceptable daily intake) of pesticides detected and the range of %ADI (the ratio of EDI to ADI) values was 0.0011~6.7087% and safety level.

• Food Science of Animal Resources
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• v.26 no.2
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• pp.263-268
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• 2006

Probiotics including Lactobacillus acidophilus, refer to a group of nonpathogenic organisms that protect the human host against gastrointestinal(GI) infections by pathogenic bacteria such as Shiga toxin-producing E. coli(STEC). In the study, the inhibitory effects of STEC ATCC 43894 adhesion by L. acidophilus A4 was investigated on the HT-29 epithelial cells. Specific proteins regulated by cell Iysates of L. acidophilus A4 on STEC ATCC 43894 were also characterized by proteomic analysis. Both cell mass and Iysate of L. acidophilus A4 have exhibited the profound inhibitory activity on the HT-29 cells(about 1.5 log scale reduction). Two-dimensional gel electrophoresis(2-DE) revealed seven proteins that were up-regulated by cell Iysates of L. acidophilus A4 and three proteins that were down-regulated. In addition, three protein spots were only detected in the presence of cell Iysates. These results suggest that inhibitory effects of STEC adhesion by L. acidophilus may be due to the regulation of specific protein of STEC.

• Journal of Embryo Transfer
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• v.27 no.4
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• pp.245-252
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• 2012

The purpose of the present study was to investigate the effect of IFN-${\tau}$ on prostaglandin synthesis, cyclooxygenase-2 (COX-2) gene expression in vitro and concentration of progesterone (P4) in endometrial cells. Epithelial and stromal cells cultured in vitro were isolated from bovine endometrium and stimulated with increasing doses of IFN-${\tau}$ (0, 0.02, 0.2 and 2 ug/ml). Human chorionic gonadotropin (hCG, 1.5 IU/ml) was used as a positive control. Prostaglandin $E_2$ and $F_{2{\alpha}}$ levels in the culture media were analyzed by enzyme immunoassays and total RNA was extracted from the cells for RT-PCR. P4 concentrations of blood samples were assayed by chemiluminescent immuno assays system. In epithelial cells, COX-2 gene expression was increased in the presence of IFN-${\tau}$ (p<0.05), but it was not significantly different in all groups of stromal cells except for 2 ug/ml IFN-${\tau}$ group (p<0.05). Although IFN-${\tau}$ did not affect $PGE_2$ and $PGF_{2{\alpha}}$ production in epithelial cells, it decreased $PGE_2$ and $PGF_{2{\alpha}}$ production significantly in stromal cells (p<0.05). In vivo experiment, blood concentration of P4 was significantly increased after addition of IFN-${\tau}$ (1 ug/ml). The results indicate that PG production was mediated by COX-2 expression in stromal cells but it was not affected in epithelial cells and this suggest that treatment of IFN-${\tau}$ could improve the implantation environment of uterine by maintenance of high P4 concentration.

• Journal of Life Science
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• v.24 no.1
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• pp.54-60
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• 2014

Aromatic hydrocarbons are toxic environmental pollutants that are detrimental to the ecosystem and human health. Among them, chlorotoluene and nitrotoluene are toxic to hydrobios and irritate the skin, eyes, and respiratory organs of humans. We herein report the development of recombinant microbial biosensors for cheap and rapid monitoring of chlorotoluene and nitrotoluene compounds. Plasmids were constructed by inserting the xylR regulatory gene for BTEX (benzene, toluene, ethylbenzene, and xylene) degradation into upstream of Po' (the DmpR activator promoter Po with the deletion of its own upstream activating sequences) or Pu (the cognate promoter of XylR)::lacZ (the ${\beta}$-galactosidase gene) and transformed into Escherichia coli $DH5{\alpha}$. In the presence of inducers, the biosensor cells immobilized in agarose developed a red color in 1-2 h due to the hydrolysis of chlorophenol red ${\beta}$-D-galactopyranoside (CPRG), a substrate of ${\beta}$-galactosidase that was expressed by the inducers. Among BTEX, high responses were specifically observed with o-, m-, p-chlorotoluene ($0.1{\mu}M-100 mM$) and o-, m-, p-nitrotoluene (0.1 mM-100 mM). Po' demonstrated higher responses than those with Pu. The biosensors immobilized in agarose showed good stability after 21 days' storage at $4^{\circ}C$, and responses in untreated wastewater spiked with chlorotoluene and nitrotoluene, suggesting they can be used to detect compounds in wastewater.

• Journal of Life Science
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• v.27 no.11
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• pp.1349-1356
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• 2017

Potassium channel openers (KCOs) produce physiological and pharmacological defense mechanisms against cell injuries caused by oxidative stress of diverse origins. Openings of mitochondrial and plasmalemmal $K^+$ channels are involved in the defense mechanisms. This study tested whether NS 1619, an opener of large-conductance BK channels, has a similar beneficial influence on the pigment epithelial cells of retinas. The human retinal pigment epithelial cell line ARPE-19 was exposed to $H_2O_2$-induced oxidative stress in the absence and presence of NS 1619. The degrees of the cells' injuries were assessed by analyzing the cells' trypan-blue exclusion abilities and TUNEL staining. NS 1619 produced remarkable protections against cell injuries caused by $H_2O_2$. It prevented apoptotic and necrotic cell deaths. The protective effect of NS 1619 was significantly diminished when the cells were treated with NS 1619 in combination with the BK channel-blocker paxilline. NS 1619 significantly ameliorated cellular ATP deprivations in $H_2O_2$-treated cells. It helped mitochondria preserve their functional integrity, which was estimated by their MTT reduction abilities and mitochondrial membrane potential. In conclusion, it was suggested that NS 1619 had a beneficial effect on mitochondria in regards to preserving their functional integrity under oxidative stress, and it produces defense mechanisms against oxidant-induced cell injuries in ARPE-19 cells.

• Clinical and Experimental Reproductive Medicine
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• v.25 no.2
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• pp.207-216
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• 1998

Certain types of somatic cells, as well as follicular cumulus cells associating with mammalian oocytes, are known to produce beneficial effects on in vitro fertilization and pre implantation development of mammalian eggs when they are present in oocyte culture medium. To investigate the nature of the effects of somatic cells, the resistance of mouse oocytes against chymotrypsin treatment was examined after culture within various cell conditioned media. When mouse oocytes matured for 17-18 hr in the presence of cumulus cells were treated with 1 % chymotrypsin, half of them remained still alive even after 240 min $(t_{50}>240.0)$. In contrast half of mouse oocytes cultured without cumulus cells underwent degeneration within 65.0 min $(t_{50}=65.0{\pm}13.2min)$ of the same treatment. To see if the effects were duc to the secretory products of cumulus cells, mouse cumulus cells were cultured for 20 hr in medium containing 0.4% BSA and the supernatant of culture medium (conditioned medium) was taken. After maturation in the cumulus cell conditioned medium, mouse oocytes exhibited $t_{50}=190.0{\pm}10.8$ min upon chymotrypsin treatment whereas half of oocytes cultured without conditioned medium degenerated within 25.5 min. Human granulosa cell conditioned medium gave similar effects such that oocytes matured in conditioned medium exhibited $t_{50}=183.3{\pm}19.1$ min while $t_50$ of control group oocytes was $60.0{\pm}6.8$ min, Oocytes matured in vero cell conditioned medium exhibited $t_{50}=196.7{\pm}8.8$ min. On the other hand, amniotic cell conditioned medium resulted in the chymotrypsin resistance of $t_{50}=80.0{\pm}8.4$ min which was not statistically different from the control value of $t_{50}=48.0{\pm}13.2$ min. Based upon these results, it is suggested that certain somatic cell types including cumulus cells might change the biochemical properties of mouse oocyte membrane during meiotic maturation as revealed by the enhanced resistance against chymotrypsin treatment. Such effects of somatic cells appear to be mediated via the secretory products rather than direct communication between somatic cells and oocytes.

• Animal cells and systems
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• v.1 no.1
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• pp.157-164
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• 1997

Ly-6E.1 antigen was proposed as a regulatory molecule of T lymphocyte activation, a hematopoietic stem cell marker, a memory cell marker, and an adhesion molecule. Though there were several reports suggesting the presence of Ly-6 ligand, the characterization of the ligand was not yet performed, As an attempt to screen the expression of Ly-6E.1 ligand, we prepared a probe for detecting Ly-6E.1 ligand by producing a fusion protein between Ly-6E.1 and $hlgC_{r1}$, A mammalian cell expression vector with Ly-6E.$1/hlgC_{r1}$ chimeric cDNA was transfected in SP2/0-Ag14 myeloma cells, and stable transfectants were selected. The fusion protein was produced as a dimer and maintained the epitopes for monoclonal antibodies specific for Ly-6E.1 and for anti-human lgG antibody. The purified fusion protein through Gammabind G column was used for FACS analyses for the expression of Ly-6E.1 ligand. The fusion protein interacted with several cell lines originating from B cells, T cells, or monocytes. The fusion Protein also strongly stained bone marrow, lymph node, and spleen cells, but thymic cells weakly, if any. The staining was more obvious in C57BL/6 $(Ly-6^b)$ than Balb/c $(Ly-6^a)$ mice. These results suggest that the interaction of Ly-6E.1 with Ly-6E.1 ligand may function both in the stem cell environment and in the activation of mature lymphocytes. The fusion protein may be a valuable tool in characterization of biochemical properties of the Ly-6E.1 ligand and, further, in isolating its cDNA.

• Journal of Korean Medical classics
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• v.28 no.4
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• pp.41-56
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• 2015

Objectives : This research is aimed to compare the visceral figures published during the Song Dynasty, and to reveal the differences between anatomical and physiological knowledge in the presence of Taoist theoretical background and in its absence. It is also aimed to discuss the theoretical differences among existing medicines. Methods : First, we will find the commonalities and the differences in the drawings describing the internal organs of a human body in the two publications, and study what made such commonalities and differences and how they affected medical theories. Secondly, we will analyze and update the internal organ data specified in the two publications as it relates to the five viscera and six bowls. Finally, we will analyze the commonalities and the differences, and study the background and the significance of medical theories. Results & Conclusions : 1. The positions of the liver and the spleen in the drawing of the Yanluozi's and Huatuo's visceral figures are inverted in the drawing of the Huatuo's visceral figures. The positions of other internal organs are similar. 2. The section of the five viscera in the Yanluozi's and Huatuo's visceral figures describes the Taoist gods necessary to build up discipline by the means of meditation. 3. In the Zhutidian's visceral theory, a belly button is recognized as a very important organ for the activity of life. 4. In the Huatuo's visceral figures, the 'Zangzhen' generated from foods is described as the energy source for the vitality of the five viscera. 5. The Huatuo's visceral figures mentioned the functionality of a gall bladder and an Tripple Energizer, which was not mentioned in the Yanluozi's and Huatuo's visceral figures. 6. Both the "Yanluozi's and Huatuo's visceral figures" and the "Huatuo's visceral figures" specify that the moisture filtered through small intestines turns into urine.

• Journal of Life Science
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• v.17 no.1 s.81
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• pp.31-38
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• 2007

Infection with virulent or attenuated Salmonella typhimuriumhas known to induce reduction in proliferative responses of spleen cells. We investigated a role of lipid A from S. typhimurium, a B cell mitogen, on proliferation of spleen cells by T cell mitogens such as concanavaline A and phytohemagglutinin under in vitro and ex vivo conditions. Lipid A alone induced proliferation of spleen cells in vitroin a dose-dependent manner. However, subsequent treatment of concanavaline A or phytohemagglutin in after lipid A treatment induced proliferation suppression of murine spleen cells in vitro and ex vivo. Removal of macrophages from spleen cells, which were obtained from a lipid A-injected mouse, restored proliferation by concanavaline A and phytohemagglutinin, indicating that macrophages appeared to play a role in lipid A-induced suppression. Secreted molecules from macrophages did not accounted for the suppression because suppressive effect was not achieved when the supernatant from macrophage-containing spleen cell culture was conditoned to macrophage-depleted spleen cell culture. Co-culture of spleen cells from lipid A-treated and - untreated mice showed proliferation suppression as increasing cell numbers of lipid A-treated mouse. These data suggested that the cell-to-cell contact of macrophage with splenic lymphocyte cells is responsible for immune responses against lipid A, which is applicable to the case of human S. typhi infection.