• Title/Summary/Keyword: Q-FISH

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Comparison of Hsp90 and CYP1A Expression Patterns by Water Temperature Stress in Atlantic Salmon (Salmo salar) (대서양 연어(Salmo salar)의 수온 스트레스에 의한 Hsp90 및 CYP1A 발현 양상 비교)

  • Kang, Han Seung;Song, Jae-Hee;Kang, Hee Woong
    • Journal of Marine Life Science
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    • v.3 no.2
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    • pp.51-58
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    • 2018
  • Variations in water temperature are known to affect almost every part of fish physiology. The rise in water temperature due to climate change can physically damage fish. This study was conducted to evaluate the health status of the Atlantic salmon (Salmo salar) at high water temperature (20℃) than the optimum water temperature (15℃). Liver tissue exerts important metabolic functions in thermal adaptation. Therefore, liver tissue was used in this study. The evaluation method is to develop the biomarker gene using NGS RNAseq analysis and to examine the expression pattern using RT-qPCR analysis. The NGS RNAseq analysis revealed 1,366 differentially expressed genes, among which 880 genes were increase expressed and 486 genes were decrease expressed. The biomarker genes are such as heat shock protein 90 alpha (Hsp90α), heat shock protein 90 beta (Hsp90β) and cytochrome P450 1A (CYP1A). The selected genes are sensitive to changes in water temperature through NGS RNAseq analysis. Expression patterns of these genes through RT-qPCR were similar to those of NGS RNAseq analysis. The results of this study can be applied to other fish species and it is considered to be useful industrially.

Distribution of Telomeric DNA in Korean Native Chicken Chromosomes (한국 재래닭 염색체의 텔로미어 분포 양상)

  • Sohn, Sea-Hwan;Cho, Eun-Jeong
    • Korean Journal of Poultry Science
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    • v.37 no.3
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    • pp.247-253
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    • 2010
  • Telomeres are nucleoprotein structures at the ends of chromosomes consisting of DNA sequences arranged in tandemly repeated units $(TTAGGG)_n$. However, this hexamers can also occur at non-telomeric sites in some avians and vertbrate. This study was carried out to present the distribution of telomeric DNA sequences in Korean Native Chicken chromosomes. The fluorescence in situ hybridization technique using a telomeric DNA probe was performed to the metaphase spreads of chicken early embryonic cells. Telomeric DNA signals were detected at the ends of chromosomes including macrochromosomes and microchromosomes. In chicken, surprisingly, chromosome 1 showed very distinct interstitial telomeric DNA hybridization patterns which located two interstitial sites in the p-arm at 1p11 and 1p23, and one in the q-arm at 1q32. In chromosome number 2 and 3 also displayed interstitial telomeric signals (ITS) in the long arms at 2q24 and 3q32, respectively. The pattern of telomeric DNA distribution in Korean Native Chicken chromosomes was in agreement with a previously reported in Gallus domesticus. The relative amount of telomeric DNA sequences in each macrochromosomes ranged from 4.6% to 16.3%. Distribution of telomeric DNAs at the end of p-arm was much more than that of q-arm in almost chicken chromosomes. The distribution of ITS in chicken chromsomes implicate to tandem chromosome fusions that might have occurred during the process of karyotype evolution.

Analysis of the Amount of Telomeric DNA and Telomerase Activity on Preimplantation Mouse Embryoic Cells (마우스 수정란의 초기 배 발생단계별 Telomeric DNA의 양적 분석과 Telomerase 활성도 분석)

  • Kang M. Y.;Han M. S.;Lee S. C.;Kim J. H.;Sohn S. H.
    • Reproductive and Developmental Biology
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    • v.29 no.1
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    • pp.1-7
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    • 2005
  • Telomeres consisting of (TTAGGG)n tandem repeat DNA sequences and associated proteins are essential for chromosome stability and related with cell senescence, apoptosis and cancer. The telomerase is a ribonucleoprotein which act as a template for the synthesis of telomeric DNA. This study was carried out to identify the distribution of telomeres on mouse chromosomes and also to analyze the amount of telomeres and telomerase activity of mouse embryos at early embryonic stages. Germ cells and early embryos from 1 cell to blastocyst stage were analyzed. The amount of telomeres was analyzed by quantitative fluorescence in situ hybridization technique(Q-FISH) using a human telomeric DNA probe, and telomerase activity was measured by telomeric repeat amplification protocol assay(TRAP). In results, the telomeres on mouse chromosomes were distributed at the ends of all autosomes and sex chromosomes. Although the quantity of telomeres varied among chromosomes, most of chromosomes had higher amount in q-arm telomeres than in p-arm telomeres. The results of Q-FISH indicated that the relative amount of telomeres of mouse embryos in each embryonic stage was approximately the same except the higher amount in blastocysts. Using TRAP assay on mouse embryos, telomerase activity was detected in all preimplantation stages from mature oocytes to blastocysts. Especially the telomerase activity was significantly increased at the morula and blastocyst stage. In conclusion, there may be a close association between the amount of telomeres and telomerase activity in early embryonic stages, and analysis of telomere quantity and telomerase activity on early development will be helpful for the investigation of embryogenesis and embryonic cell differentiation in mice.

Effect of Water Temperature and Body Weight on Oxygen Consumption Rate of Starry Flounder Platichthys stellatus (강도다리 Platichthys stellatus의 산소 소비율에 미치는 수온과 체중의 영향)

  • Oh, Sung-Yong;Jang, Yo-Soon;Noh, Choong Hwan;Choi, Hee Jung;Myoung, Jung-Goo;Kim, Chong-Kwan
    • Korean Journal of Ichthyology
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    • v.21 no.1
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    • pp.7-14
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    • 2009
  • The effect of water temperature (T) and body weight (W) on oxygen consumption of fasted starry flounder Platichthys stellatus was investigated in order to assess the metabolic response of this species at given conditions. The oxygen consumption rate (OCR) was measured under six different water temperatures (4, 7, 10, 13, 16 and $19^{\circ}C$) and at two different body weights (mean weight of fry group : 1.5 g; fingerling group : 37.4 g) at an interval of 5 minutes for 24 hours using a continuous flow-through respirometer. In each treatment three replicates were set up and a total 540 fish in fry groups and 90 fish in fingerling groups were used. The OCRs increased with increase of water temperature in both groups (p<0.001). Mean OCRs at 4, 7, 10, 13, 16 and $19^{\circ}C$ were 1386.0, 1601.7, 1741.0, 1799.2, 2239.1 and $2520.3mg\;O_2\;kg\;fish^{-1}\;h^{-1}$ in fry groups, and 83.8, 111.4, 126.3, 147.1, 187.7 and $221.3mg\;O_2\;kg\;fish^{-1}\;h^{-1}$ in fingerling groups, respectively. The OCRs decreased with increasing body weights at six different water temperatures (p<0.001). The relationship between water temperature and body weight is described by the following equation : OCR=1520.91+40.85T-49.22W ($r^2=0.95$, p<0.001). The energy loss by metabolic response increased with an increase in water temperature and a decrease in body weight (p<0.001). Mean energy loss rates by oxygen consumption at 4, 7, 10, 13, 16 and $19^{\circ}C$ were 907.9, 1046.5, 1141.6, 1177.0, 1467.3 and $1650.1kJ\;kg\;fish^{-1}\;d^{-1}$ in fry groups and 54.8, 73.0, 82.9, 96.2, 122.9 and $144.6kJ\;kg\;fish^{-1}\;d^{-1}$ in fingerling groups, respectively. The $Q_{10}$ values of fingerling groups were higher than those of fry groups at given temperature ranges. The $Q_{10}$ values at $4{\sim}7^{\circ}C$, $7{\sim}10^{\circ}C$, $10{\sim}13^{\circ}C$, $13{\sim}16^{\circ}C$ and $16{\sim}19^{\circ}C$ were 1.62, 1.32, 1.12, 2.07 and 1.48 in fry groups, and 2.59, 1.52, 1.67, 2.25 and 1.73 in fingerling groups, respectively.

Antibacterial activity of new quinolones against Edwardsiella tarda isolated from eel tanks (양만장에서 분리된 Edwardseilla tarda균에 대한 new quinolone계 약물의 항균작용)

  • Choi, Min-Soon;Kim, Kyong-Ho;Seo, Young-Ho;Choi, Sang-Hoon;Park, Kwan-Ha
    • Journal of fish pathology
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    • v.9 no.2
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    • pp.185-193
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    • 1996
  • Extensive acquirement of drug resistance to traditional antibacterial agents poses a serious problem to eel aquaculturists. To collect the basic information for new drug development in the future, we assessed the in vitro antibacterial efficacy of 14 new quinolones with 75 isolates of Edwardsiella tarda from local aquaculture tanks of Anguilla japonica. Of all tested quinolones under development or marketed for human use, DU-6859 was most potent with its $MIC_{50}$ value of $0.05{\mu}g$/ml in broth microdilution assay. The drugs whose $MIC_{50}$ values ranged from 0.2 to $0.78{\mu}g$/ml were T-3762, Bay-y3118, ciprofloxacin, norfloxacin, ofloxcin and tosufloxacin. The weakest group of drugs, with their $MIC_{50}$ being 1.56-$3.13{\mu}g$/ml, were difloxacin, sparfloxacin, fleroxacin, Q-35, amifloxacin, lomefloxacin and enoxacin. The number of resistant strains, when arbitrarily defined with their MICs of $\geq6.25{\mu}g$/ml, was : 3 to T-3762, 3 to Bay-y3118, 44 to difloxacin, 16 to sparfloxacin, 13 to ciprofloxacin, 19 to fleroxacin, 36 to Q-35). 31 to amifloxacin, 5 to norfloxacin, 13 to ofloxacin, 31 to lomefloxacin, 41 to enoxacin, 12 to tosufloxacin and 0% to DU-6859, respectively. This information can be taken into consideration for the future development of fisheries antibacterial quinolones.

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A chromosome 1q44 deletion in a 4-month-old girl; The first report in Korea

  • Cho, Joo Hyun;Song, Eun Song;Kim, Hee Na;Oh, Burm Seok;Choi, Young Youn
    • Clinical and Experimental Pediatrics
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    • v.57 no.6
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    • pp.292-296
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    • 2014
  • The deletion of the distal long arm of chromosome 1 is associated with a characteristic facial appearance and a pattern of associated malformations. Characteristic manifestations include a round face with prominent 'cupid's bow' and downturned corners of the mouth, thin vermilion borders of lips, a long upper lip with a smooth philtrum, a short and broad nose, epicanthal folds, apparently low-set ears, micrognathia, microcephaly, abnormal hands and feet, variable cardiac or genital anomalies, moderate to severe mental retardation, and growth retardation. Using fluorescent in situ hybridization (FISH) analysis to map precisely the deletion, we present a case of chromosome 1q44 deletion with craniofacial characteristics, multiple congenital anomalies, and growth and psychomotor retardation. In comparison with other reported cases of 1q43-44 deletion, the subject does not show hydrocephalus, seizure, syn- or polydactyly of hands, and a urogenital anomaly. However, an arachnoid cyst, pinpoint dimple on the midline of the forehead, a right-sided supernumerary nipple and auricular pit, polydactyly of the right foot, adducted thumb, and flexion restriction of the proximal interphalangeal joint with a simian line in both hands were observed additionally.

Biofilm Processes for Volume Decrease in Recirculating Water Treatment Systems for Aquaculture

  • Kim Jeong-Sook;Yoon Gil-Ha;Ghim See-Jun;Kang Lim-Seok;Lee Byung-Hun
    • Fisheries and Aquatic Sciences
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    • v.1 no.2
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    • pp.242-249
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    • 1998
  • The engineering aspect of water treatment processes in the recirculating aquaculture system was studied. To recycle the water in the aquaculture system, a wastewater treatment process was required to maintain high water quality for the growth and health of the cultured fish. In this study, three different biofilm processes were used to reduce the concentration of organic matters and ammonia from the recirculating water - two phase fluidized bed, three phase fluidized bed, and trickling filter. The objectives of this research were to evaluate the optimum treatment conditions of the biofilm processes for the recirculating aquaculture system, and thereby reduce the volume of biofilm processes, which are commonly used for the recycle water treatment processes for aquaculture. The result of this study showed that the removal efficiency of organic matters by trickling filter was found to be lower than that of the fluidized bed. In the trickling filter system, anthracite showed better organic removal efficiency than crushed stone as a media. In the two phase fluidized bed, the maximum removal efficiency of either organics or ammonia was obtained when both the packing rate of media was maintained to $40\%$ of total reactor depth excepting sediment zone and the bed expansion rate was maintained to $100\%$. When 100 tilapia (Oreochromis niloticus) of each average 200g was reared, the pollutant production rate was 0.07g $NH_4\;^+-N/kg$ fish/day and 0.06g P04-3-P/kg fish/day, and sludge production rate was 0.39 g SS/kg fish/day. In the two phase and three phase fluidized bed, the volume of water treatment tank could be calculated from an empirical equation by using the relationship between the influent COD to $NH_4\;^+-N$ ratio (C/N, -), media concentration (Cm, g/L), influent ammonia nitrogen concentration (Ni, mg/L), effluent ammonia nitrogen concentration (Ne, mg/L), bed expansion rate $(E,\;\%)$, and influent flowrate $(Q,\;m^3/hr)$. The empirical equation from this study is $$V_2\;=\;10^{3.1279}\;C/N^{3.5461}\;C_m\;^{-3.7473}\;N_i\;^{4.6477}\;E^{0.0326}\;N_e\;^{-0..8849}\;Q\;(Two\;Phase\;FB) V_3\;=\;10^{11.7507}\;C/N^{-1.2330}\;C_m\;^{-6.5715}\;N_i\;^{1.5091}\;N_e\;^{-1.8489}\;Q (Three\;Phase\;FB)$$

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Gene Expression Profile in Iprobenfos Exposed Medaka Fish by Microarray Analysis

  • Woo, Seon-Ock;Son, Sung-Hee;Ryu, Jae-Chun;Yum, Seung-Shic
    • Molecular & Cellular Toxicology
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    • v.4 no.2
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    • pp.132-137
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    • 2008
  • Differential gene expression profiling was carried out in the hepatic tissue of medaka fish, Oryzias latipes, after exposure to an organophosphorus pesticide (OPP), Iprobenfos (IBP), a widely used pesticide in agri- and fish-culture, using a medaka cDNA micro array. Twenty six kinds of differentially expressed candidate genes, with 15 and 11 induced and repressed in their gene expressions, respectively, were associated with cytoskeleton (3.8%), development (7.7%), immune (7.7%), metabolism (30.8%), nucleic acid/protein binding (42.3%) and reproduction (7.7%). Of these genes, changes at the transcription level of five were re-evaluated by real-time quantitative PCR (qRT-PCR). Considering the known function of authentic genes, the effects of IBP on the biological activity and pathological aspects in medaka fish were discussed. The identified genes could be used as molecular biomarkers for biological responses to OPPs contamination in an aquatic environment.

Anaesthetic Tolerance of Juvenile Black Rockfish Sebastess schlegeli, Produced for Wild Stock Enhancement

  • Son, Maeng-Hyun;Park, Min-Woo;Myeong, Jeong-In;Kim, Dae-Jung;Kim, Byoung-Hak;Jo, Q-Tae;Jeon, Im-Gi
    • Ocean and Polar Research
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    • v.23 no.3
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    • pp.285-290
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    • 2001
  • The strength of juvenile black rockfish, Sebastes schlegeli, raised in different hatcheries for wild stock enhancement was evaluated in terms of resistance to an anesthetizing agent, tricaine methane-sulfonate (MS-222), and exposure to drying. The working dosage of MS-222 varied significantly with fish size and hatchery population. Smaller fish were less resistant to the chemical than larger ones. MS-222 effects also differed with fish growth history. The fish cultured in embanked populations showed stronger resistance, earlier recovery, and lower mortality, compared to those cultured in land-based tanks or collected from wild stocks. Similar results were seen in juveniles challenged to dry exposure. These results suggest that an embanked population of black rockfish is more resistant to anesthetic stress, expressed as anesthesia recovery and mortality, and that this population is healthier than others.

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Effects of dietary lipid sources on apoptotic and immune gene expression in head kidney of olive flounder (Paralichthys olivaceus)

  • Hur, Deokhwe;Lee, Sang-Min;Hong, Suhee
    • Journal of fish pathology
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    • v.25 no.3
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    • pp.199-210
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    • 2012
  • It can be hypothesized that dietary fatty acids can modulate immune responses in fish by inducing apoptosis of immune cells since dietary polyunsaturated fatty acid (PUFA) increase apoptosis by oxygen radicals generated by peroxidation. Thus we examined the effects of deferent dietary lipid sources such as squid liver oil (FO), linseed oil (LO) and soybean oil (SO) on oxidation (Cytochrome C oxidase; COS), apoptosis (TNF-${\alpha}$ Scinderin like) and immune (IL-$1{\beta}$ and NKEF) gene expression in the main immune organ (head kidney) in olive flounder (Paralichthys olivaceus) by Q-PCR analysis after feeding diets containing each oil (5%) for 15 weeks. Linseed oil and soybean oil were chosen to compare n-3 or n-6 enriched vegetable oils, respectively. Consequently, COS, TNF-${\alpha}$ and Scinderin like gene expression was increased in SO group, indicating the induction of oxidation and apoptosis. Meanwhile, no significant difference was found in immune gene expression. In conclusion vegetable oils containing n-3 PUFA like linseed oil seems to be more suitable lipid source than soybean oil for replacement of fish oil in flounder since n-6 PUFA in SO leads to activation of apoptosis pathways within the cellular damage in head kidney.