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Transcriptional Activity of an Estrogen Receptor β Subtype in the Medaka Oryzias dancena

  • Maeng, Sejung;Yoon, Sung Woo;Kim, Eun Jeong;Nam, Yoon Kwon;Sohn, Young Chang
    • Development and Reproduction
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    • v.23 no.4
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    • pp.333-344
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    • 2019
  • In vertebrate reproductive system, estrogen receptor (ER) plays a pivotal role in mediation of estrogenic signaling pathways. In the present study, we report the cDNA cloning, expression analysis, and transcriptional activity of ERβ1 subtype from medaka Oryzias dancena. The deduced O. dancena ERβ1 (odERβ1; 519 amino acids) contained six characteristic A/B to E/F domains with very short activation function 2 region (called AF2). A phylogenetic analysis indicated that odERβ1 was highly conserved among teleost ERβ1 subgroup. A conventional RT-PCR revealed that the odERβ1 transcripts were widely distributed in the multiple tissues, the ovary, brain, gill, intestine, kidney, and muscle. Further, the relatively higher odERβ1 expressions in the ovary and brain were clearly reproduced in RT-qPCR assay. When HA-fused odERβ1 expression vector was transfected into HEK293 cells, an immunoreactivity for odERβ1 was mainly detected in the nucleus part. Finally, an estrogen responsive element driven luciferase reporter assays demonstrated that the transcriptional activity of odERβ1 significantly increased by estradiol-17β (E2) in a dose dependent manner (p<0.05). However, fold-activation of odERβ1 in the presence of E2 was markedly weak, when it compared with those of O. latipes ERβ1. Taken together, these data suggest that odERβ1 represents a functional variant of teleost ERβ subtype and provides a basic tool allowing future studies examining the function of F domain of ERβ1 subtype and expanding our knowledge of ERβ evolution.

Effect of Water Potential on Mycelial Growth and Production of Sclerotia of Sclerotium cepivorum (Sclerotium cepivorum의 균사생장 및 균핵 생성에 대한 수분압의 영향)

  • Lee, Yong-Hoon;Lee, Du-Ku;Lee, Wang-Hyu
    • Korean Journal Plant Pathology
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    • v.13 no.4
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    • pp.200-204
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    • 1997
  • The effect of water potential ($\psi_W$) and temperature on mycelial growth and sclerotial production of Sclerotium cepivorum was determined in potato dextrose agar(PDA) and potato dextrose broth (PDB) adjusted to different $\psi_W$ with NaCI, KCI, sucrose or polyethylene glycol (PEG) at 15, 20 and $25^{\circ}C$. the growth of mycellium was not significantly af. fected by $\psi_W$ values between -1,970 and -2,240J/Kg, but severely decreased lower than -2,240J/Kg. Dry weight was slightly increased at $\psi_W$values between -450 and -2,240 J/Kg. The reduction of dry weight wasslower than the reduction of mycelial growth as the $\psi_W$ decreased. The mycelial growth and dry weight were more severely influenced on PEG amended media than on other osmotica amended media. About 50% reduction of mycelial growth and dry weight was occurred about -1,000 and -2240 J/Kg, respectively. The production of sclerotion of sclerotial production occurred between -450 and -810 J/Kg. Sclerotium was not produced lower than -2,240 J/Kg. Mycelial growth and sclerotial production was better at $25^{\circ}C$ as the $\psi_W$ decreased than at $20^{\circ}C$ which is optimal temperaturein the undmended media. The influence of $\psi_W$ on mycelial growth and sclerotial production of S. cepivorum adjusted with NaCl, KCI sucrose or polyethylene glycol showed similar patterns.

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Biodegradation of Biphenyl by Sphingbium yanoikuyae BK-10 (Biphenyl의 Sphingobium yanoikuyae BK-10에 의한 분해 특성)

  • Lee Jung-Bok;Kim Dong-Geol;Choi Chung-Sig;Sohn Ho-Yong;Kim Jang-Eok;Kwon Gi-Seok
    • Microbiology and Biotechnology Letters
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    • v.34 no.2
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    • pp.174-179
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    • 2006
  • Bacterium capable of using biphenyl as a sole source of carbon and energy were isolated from soil, and based on the results of 16S rDNA sequence, strain BK10 identified as a Sphingobium yanoiktiyae. The optimum cultural conditions were as follows; $NH_4NO_3$ 1g, $K_2HPO_4$ 1g, $MgSO_4{\cdot}7H_2O$ 0.5g, $CaCO_3$ 0.2 g per 1 liter of distilled water. The Sphingobium yanoikuyae BK10 strain was completely utilized biphenyl in mineral salt media containing biphenyl at concentration 500 $\mu$g/ml of biphenyl as a sole carbon and energy source within 48 hours. Optimumal pH and temperature for biphenyl degradation and cell growth of strains were 6.0$\sim$8.0 and 20$\sim$50$^{\circ}C$, respectively. Especially, at 30$^{\circ}C$, cell-growth were higher than other temperature. Cell grown on biphenyl has been shown to have a higher removal rate for biphenyl than grown on sucrose. This study shows that Sphingobium yanoikuyae BK10 strain had a high biodegradation capability of biphenyl and can be simulate a candidate compounds the bioremediation of PCBs (Polychlorinated biphenyl) contaminant soil and water.

The Effect of Dissolved Oxygen and Salinity on Oxygen Consumption and Ammonia Excretion in the Mysid, Neomysis awatschensis (곤쟁이 (Neomysis awatschensis)의 산소소비와 질소배설에 미치는 염분 및 용존산소의 영향)

  • SHIN Yun Kyung;LEE Jung Ah;CHIN Pyung
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.33 no.2
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    • pp.110-114
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    • 2000
  • Effects of the combinations of six oxygen concentrations ($control, 0.6, 1.0, 2.0, 3.4 and 7.4 mg DO/l$) and two salinity levels ($20{\%_{\circ}} and 32{\%_{\circ}}$) on the rates of oxygen consumption, ammonia excretion and mortality of the mysid, Neomysis awatschensis were tested at $20{\circ}C$. The lethal level ($96 hr-LC-(50)$) of dissolved oxygen for mysid at $20{\%_{\circ}} and 32{\%_{\circ}} were 2,20 mg DO/l and 1.60 mg DO/l$ respectively, and all mysids died within $24hr at 0.6 mg DO/l$. Oxygen consumption rate of mysid was increased with dissolved oxygen increase at $20{\%_{\circ}} and 32{\%_{\circ}}$, but ammonia excretion rate was high af $1.0 mg DO/l$ during 96h exposure to DO concentration, and significantly greater in $20{\%_{\circ}} than 32{\%_{\circ}}$. $O:N$ ratio of mysid exposed during 96hr with salinity anil dissolved oxygen was below $10 at 20{\%_{\circ}} and 1,0{\~}2.0 mg DO/l, and was 4.4 at 32{\%_{\circ}} and 1.0 mg DO/l$. These results indicated that mysids were capable of changing their energy substrate in response to salinity and DO changes, and obtaining energy from proteins.

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Influence of Dietary Supplemental Sardine Oil on Storage and Processing Characteristics of Broiler (정어리유의 급여가 계육의 저장성 및 가공 특성에 미치는 영향)

  • 박창일;김영길;김영직
    • Food Science of Animal Resources
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    • v.22 no.1
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    • pp.8-12
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    • 2002
  • This study was conducted to investigate the influence of dietary sardine oil on storage and processing characteristics in meat sample of chicken meat. Broilers were randomly assigned to one of four dietary treatments: 1) Control(commercial feed) 2) T1(commercial feed supplemented with 1% sardine oil) 3) T2(commercial feed with 2% sardine oil) and 4) T3(commercial feed with 4% sardine oil). They were fed one of the experimental diets for five weeks and slaughtered. After that, the meat samples were vacuum packaged and stored at 4$\pm$1$\^{C}$. The storage and processing characteristics were analyzed for meat samples stored over a period of 0, 1, 3, 7 and 10 days. The pH of all treatments significantly increased during the storage periods (p<0.05). The TBARS(thiobarbituric acid reactive substances) af all treatments were significantly increased as storage period extended (p<0.05). After 1 days, the TBARS of treatment groups were significantly higher than that of the control (p<0.05). The T3 showed the highest TBARS among all treatments (p<0.05). The VBN(volatile basic nitrogen) of all treatments significantly increased during storage period (p<0.05). However, the VBN was not significantly different between control and treatment groups. The WHC(water holding capacity) and heating loss were significantly increased in both control and treatment groups during storage (p<0.05) and however, WHC was not significantly different among 3 treatment. The heating loss tended to increase in treatment groups compared to the control.

Detection of Human Taurine Transporter and Production of Monoclonal Antibody

  • An, Hye-Suk;Han, Hee-Chang;Lee, Sun-Min;Park, Taesun;Park, Kun-Koo;Kim, Ha-Won
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 2001.11a
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    • pp.102-102
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    • 2001
  • Taurine (2-ethaneaminosulfonic acid) is one of the major intracellular ${\beta}$ -amino acids in mammals and is required for a number of biological processes including membrane stabilization, osmoregulation, antioxidation, detoxification, modulation of calcium flux and neurornodulation. The taurine transporter (TAUT) which contains 12 hydrophobic membrane-spanning domains has been cloned from dog kidney, rat brain, mouse brain, human thyroid, placenta and retina. In this study, The TAUT cDNA from the human intestinal epithelial cell, HT-29 was cloned and sequenced. Reverse-transcription polymerase chain reaction (RT-PCR) was performed to amplify partial cDNA encoding human intestinal TAUT. The coding region of the PCR product was 732 bp long. The primers were designed to encode highly conserved amino acid sequences near the transmembrane domains III (IPYFIFLF) and Ⅵ (KYKYNSYR) both in human and mouse. The TAUT cDNA amplified was ligated into the pGEX 4T-1 expression vector. The resulting sequence of human intestinal TAUT cDNA (Accession number of NCBI Genebank is AF346763) was identical to the sequences of the TAUTs previously determined in the human placenta and retina except 3 base pairs from that of the reported human thyroid. TAUT specific antibodies were generated to use them as biological tools in the studies of the biological role of TAUT. Peptides of 149-162 amino acid residue (14 amino acids) of the TAUT were synthesized. The synthetic peptide used in this study was LFQSFQKELPWAHC. This region was chosen not only to avoid putative glycosylation sites but also to exclude regions of known homology with GABA transporters in the extracellular hydrophilic domains. The synthetic peptide, TAUT-1 was conjugated with carrier protein, kehole lympet hemocyanin (KLH) to use as an antigen. When used for immunization on a rabbit to produce polyclonal antiserum, the conjugates elicited high -titered specific anti-TAUT-1 antibodies, which reacted well with the ovalbumin (OVA) conjugated peptides in ELISA. The KLH-conjugated peptide was also used as immunizing antigen in BALB/c mice to produce TAUT specific monoclonal antibodies. From the culture supernatant of the hybridoma, the specificity of anti-TAUT-1 monoclonal antibodies was confirmed by ELISA. Further applications of more tools in TAUT expression analysis will be performed such as western blotting and flow cytometry.

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Effects of Ginseng Saponin and Its Related Materials on Aflatoxin Production by Aspergillus parasiticus NRRL2999 in Synthetic Medium (합성 배지에서 Aspergillus parasiticus의 Aflatoxin 생성에 미치는 인삼 saponin과 그 관련물질의 영향)

  • 전홍기;조영배;박건영
    • Korean Journal of Microbiology
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    • v.24 no.4
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    • pp.352-356
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    • 1986
  • A study was carried out to determine the effect of ginseng saponin an its related materials on aflatoxin production by Aspergillus parasiticus NRRL2999 in glucose-salts(GS) medium. Maximal growth of the mold and AF froduction in the medium occurred after 5 and 9 days at $28^{\circ}C$, respectively. When various concentrations of saponin added to the medium aflatoxin synthesis were significantly reduced (p<0.05) compared to the control after 9 days at $28^{\circ}C$. 0.05% of saponin inhibited aflatoxin production most effectively in the low concerntrations of saponin (0.01-0.2%) and the toxin synthesis reduced with an increasing concentrations of saponin in the high concentrations (0.03-5.0%). Various concentrations (0.01-1.0%) of saponin diol and triol in the media also caused to reduce aflatoxin synthesis by the mold (p<0.05). All saponin fractions were found to decrease aflatoxin production significantly. Saponin fraction numbers of 1,2,4 and 6 were shown to reduce aflatoxin production effectively, and the number 1 was the most effective. Addition of 0.05% of nucleic acid related materials to the medium reduced aflatoxin production (p<0.05). Aflatoxins could not be found in broth at all, but in mycelia when 0.05% of caffeine was added to the medium. Aflatoxin synthesis was well correlated with total lipid synthesis, growth and glucose uptake. When aflatoxin synthesis inhibited (5.0% of saponin) both total lipid synthesis and growth were stimulated and the efficiency of glucose utilization was reduced.

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Effect of the Addition of "Royal Jelly" on Post-thaw Viability and Longevity of Canine Spermatozoa (Royal Jelly 첨가가 동결융해 후 개 정자의 활력도 및 생존성에 미치는 영향)

  • 공일근;조성균
    • Journal of Embryo Transfer
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    • v.16 no.1
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    • pp.53-60
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    • 2001
  • This study was conducted to evaluate whether \"Royal jelly\" (RJ) added to Tris-buffer dilute contributed to supporting post-thaw viability and longevity of frozen canine spermatozoa. Two Japanese spitzs (2 to 4 years of age) were used as a semen donor. Semen was collected by manual masturbation and separated into 3 fractions. Only the sperm-rich fraction having sperm motility of more than 70%, containing sperm concentration of 2~4$\times$10$^{8}$ cells/ml and having dead or abnormal spermatozoa of less than 15% was used for the experiment. Each ejaculated semen was centrifuged at 400 $\times$ g for 5 min and then diluted in a Tris-buffer supplemented with 20 ml egg yolk (Ext I), 4% glycero1 and 1% Equex STM Paste (Ext II) or g1ycero1, Equex STM paste and RJ of various concentrations (Ext II-RJ). After freezing and thawing, viability of spermatozoa in Ext II -RJ containing 1% RJ immediately after thawing (67.5$\pm$9.6) was significantly lower than that of Ext II , Ext II -RJ containing 0.01 or 0.1% RJ (77.5$\pm$12.5, 78.7$\pm$8.2 and 80.0$\pm$6.3). However, Ext II-RJ containing 0.1% RJ yielded higher viability than Ext II, Ext II-RJ containing 0.01% at or 1% 1 h after thawing (69.5$\pm$8.1 vs. 55.0$\pm$12.9, 57.5$\pm$9.6 and 41.5$\pm$12.6; P<0.05). At 1 h after thawing, the viability of spermatozoa thawed in 7$0^{\circ}C$ (68.8$\pm$12.5) was significantly higher than that of spermatozoa thawed in 38$^{\circ}C$ (48.8$\pm$16.3), although there was no difference in the viability between both groups immediately after thawing (77.5$\pm$9.6 and 81.3$\pm$8.1). Post-thaw viability and longevity of post-thaw spermatozoa in Ext II-RJ containing 0.1% RJ was higher in those in Ext II at 1 h (65.0$\pm$12.9 vs. 42.5$\pm$12.6), 2 h (52.5$\pm$12.6 vs. 27.5$\pm$17.1) and 3 h (40.0$\pm$14.1 vs. 20.0$\pm$12.1) after thawing. These results indicated that addition of 0.1% af to Tris-buffer enhanced post-thaw viability and longevity of canine spermatozoa and this additive can be used for increasing the possibility of collision between spermatozoa and ova during insemination.emination.

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Effects of Microstructures on the Toughness of High Heat Input EG Welded Joint of EH36-TM Steel (EH36-TM강의 대입열 EGW 용접부 저온 인성에 미치는 미세 조직의 영향)

  • Choi, Woo-Hyuk;Cho, Sung-Kyu;Choi, Won-Kyu;Ko, Sang-Gi;Han, Jong-Man
    • Journal of Welding and Joining
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    • v.30 no.1
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    • pp.64-71
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    • 2012
  • The characteristics of high heat input (342kJ/cm) EG (Electro Gas Arc) welded joint of EH36-TM steel has been investigated. The weld metal microstructure consisted of fine acicular ferrite (AF), a little volume of polygonal ferrite (PF) and grain boundary ferrite (GBF). Charpy impact test results of the weld metal and heat affected zone (HAZ) met the requirement of classification rule (Min. 34J at $-20^{\circ}C$). In order to evaluate the relationship between the impact toughness property and the grain size of HAZ, the austenite grain size of HAZ was measured. The prior austenite grain size in Fusion line (F.L+0.1 mm) was about $350{\mu}m$. The grain size in F.L+1.5 mm was measured to be less than $30{\mu}m$ and this region was identified as being included in FGHAZ(Fine Grain HAZ). It is seen that as the austenite grain size decreases, the size of GBF, FSP (Ferrite Side Plate) become smaller and the impact toughness of HAZ increases. Therefore, the CGHAZ was considered to be area up to 1.3mm away from the fusion line. Results of TEM replica analysis for a welded joint implied that very small size ($0.8\sim1.2{\mu}m$) oxygen inclusions played a role of forming fine acicular ferrite in the weld metal. A large amount of (Ti, Mn, Al)xOy oxygen inclusions dispersed, and oxides density was measured to be 4,600-5,300 (ea/mm2). During the welding thermal cycle, the area near a fusion line was reheated to temperature exceeding $1400^{\circ}C$. However, the nitrides and carbides were not completely dissolved near the fusion line because of rapid heating and cooling rate. Instead, they might grow during the cooling process. TiC precipitates of about 50 ~ 100nm size dispersed near the fusion line.

NDP Kinases Suppressed Bax-Dependent Apoptosis in Yeast System

  • K. C. Hwang;D. W. Ok;D. N. Kwon;H. K. Shin;Kim, J. H.
    • Proceedings of the KSAR Conference
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    • 2001.03a
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    • pp.52-52
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    • 2001
  • Many nucleoside diphosphate (NDP) kinases are ubiquitous enzymes responsible for the exchange of ${\gamma}$-phosphates between tri- and diphosphonucleosides. The catalytic Many nucleoside diphosphate (NDP) kinases are ubiquitous enzymes responsible for the exchange of ${\gamma}$-phosphates between tri- and diphosphonucleosides. The catalytic reaction follows a ping-pong mechanism in which the enzyme is transiently phosphorylated on a histidine residue conserved in all nucleoside diphosphate kinases. Beside their role in nucleotide synthesis, these enzymes present additional functions, possibly independent of catalysis, in processes such as differentiation, cell growth, tumor progression, metastasis and development. To clone murine nm23-M5, several expressed sequence tags (ESTs) of the GenBank data base, selected according to their homology to nm23-H5 cDNA, reconstituted a complete open reading frame (GenBank AF222750). To test whether murine NDPKs (1, 2, 3, 4, 5, and 6) can inhibit Bax-mediated toxicity in yeast, co-transformation was performed respectively. The yeast S.cerevisiae was transformed with a copy expression plasmid containing the histidine selection marker and expressing murine Bax under the control of a galactose-inducible promoter. Several clones were selected and found to be growth inhibited when Bax expression was induced with galactose. A representative clone was transformed again with a copy expression plasmid containing the tryptophane selection marker and expressing either murine Bcl-xL or NDPK under the control of a galactose-inducible promoter. Several subclones of the double-transformants were selected and characterized. The ability of Bcl-xL and NDPKs to suppress Bax-mediated toxicity was determined by growing yeast cells overnight in galactose media and spot-testing on galactose plates starting with an equal number of yeast cells as determined by taking the OD$_{600}$. Ten-fold serial dilutions were used in the spot-test. Plates were grown at 3$0^{\circ}C$ for 2-3 days. All murine NDPKs suppressed Bax dependent apoptosis. Futher study will be peformed whether Bax-toxicity inhibition was caused by NDP kinase activity or additional function.n.

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