• Title/Summary/Keyword: Q-FISH

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Molecular Characterization of the HERV-W Env Gene in Humans and Primates: Expression, FISH, Phylogeny, and Evolution

  • Kim, Heui-Soo;Kim, Dae-Soo;Huh, Jae-Won;Ahn, Kung;Yi, Joo-Mi;Lee, Ja-Rang;Hirai, Hirohisa
    • Molecules and Cells
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    • v.26 no.1
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    • pp.53-60
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    • 2008
  • We characterized the human endogenous retrovirus (HERV-W) family in humans and primates. In silico expression data indicated that 22 complete HERV-W families from human chromosomes 1-3, 5-8, 10-12, 15, 19, and X are randomly expressed in various tissues. Quantitative real-time RT-PCR analysis of the HERV-W env gene derived from human chromosome 7q21.2 indicated predominant expression in the human placenta. Several copies of repeat sequences (SINE, LINE, LTR, simple repeat) were detected within the complete or processed pseudo HERV-W of the human, chimpanzee, and rhesus monkey. Compared to other regions (5'LTR, Gag, Gag-Pol, Env, 3'LTR), the repeat family has been mainly integrated into the region spanning the 5'LTRs of Gag (1398 bp) and Pol (3242 bp). FISH detected the HERV-W probe (fosWE1) derived from a gorilla fosmid library in the metaphase chromosomes of all primates (five hominoids, three Old World monkeys, two New World monkeys, and one prosimian), but not in Tupaia. This finding was supported by molecular clock and phylogeny data using the divergence values of the complete HERV-W LTR elements. The data suggested that the HERV-W family was integrated into the primate genome approximately 63 million years (Myr) ago, and evolved independently during the course of primate radiation.

External Attachment of Pop-up Satellite Archival Tag (PSAT) and Water Temperature Affect Oxygen Consumption Rate of the Olive Flounder Paralichthys olivaceus (넙치(Paralichthys olivaceus) 산소 소비율에 미치는 Pop-up Satellite Archival Tag (PSAT) 체외 부착과 수온의 영향)

  • Geun Su Lee;Pil Jun Kang;Hye Mi Park;Sung-Yong Oh
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.56 no.5
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    • pp.660-666
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    • 2023
  • This study aimed to examine the effect of external pop-up satellite archival tags (PSATs) attachment and water temperature on the oxygen consumption rate (OCR) of the olive flounder (mean body weight 2281.7 g). The OCRs of fish were measured under conditions of three different water temperature conditions (15, 20, and 25℃) and two different tagging methods [non-tagging, control; bio-logger external attachment with a miniature PSAT (dummy mrPAT), BEA] using a closed flow-through respirometer. The OCRs of fish linearly increased with the increase in water temperature in both the control and BEA (P<0.001); however, the OCRs of BEA were approximately 1.8-1.9 times lower than those of the control at each water temperature (P<0.001). The Q10 values of the control and BEA were the highest in the water temperature range of 15 to 20℃, but sensitivity to water temperature changes was higher in BEA than in the control. The metabolic energy loss rate (MEL) of fish increased with increasing water temperature regardless of external tagging, but the MEL of the control was higher than that of BEA (P<0.001). These results demonstrate that OCR, thermal sensitivity, and energy expenditure are all affected in adult olive flounder with external PSAT attachment.

Effects of Dietary Supplementation of Spirulina and Quercetin on Growth, Innate Immune Responses, Disease Resistance Against Edwardsiella tarda, and Dietary Antioxidant Capacity in the Juvenile Olive Flounder Paralichthys olivaceus

  • Kim, Sung-Sam;Rahimnejad, Samad;Kim, Kang-Woong;Lee, Bong-Joo;Lee, Kyeong-Jun
    • Fisheries and Aquatic Sciences
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    • v.16 no.1
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    • pp.7-14
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    • 2013
  • A 10-week feeding trial was conducted to examine the effects of dietary spirulina and quercetin on growth, innate immunity, disease resistance and dietary antioxidant capacity in the juvenile olive flounder. Triplicate groups of fish (initial body weight, $2.9{\pm}0.01g$) were fed one of isonitrogenous (48% crude protein) and isocaloric (17.4 MJ/kg DM) experimental diets containing 0% spirulina (as a control), 3.4% spirulina, or 6.8% spirulina with or without supplementation of 0.5% quercetin (designated as CON, SP3.4, SP6.8, and SP6.8 + Q, respectively) at a rate of 3% body mass twice daily. Higher dietary antioxidant capacity was found with spirulina supplementation, and the highest value (P < 0.05) was obtained with SP6.8 + Q diet. At the end of the feeding trial, no significant effects were observed on growth performance, body composition and disease resistance against Edwardsiella tarda. Lysozyme activity was significantly increased by spirulina supplementation (P < 0.05), and the highest value was observed in the group fed SP6.8 + Q diet. Also, significantly higher respiratory burst activity (P < 0.05) was found in SP3.4 group. According to the results of this study, dietary supplementation of 3.4% spirulina may enhance innate immunity of olive flounder.

Preliminary Application of Molecular Monitoring of the Pacific Herring (Clupea pallasii) Based on Real-time PCR Assay Utilization on Environmental Water Samples

  • Kim, Keun-Yong;Heo, Jung Soo;Moon, Seong Yong;Kim, Keun-Sik;Choi, Jung-Hwa;Yoo, Joon-Taek
    • Korean Journal of Ecology and Environment
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    • v.54 no.3
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    • pp.209-220
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    • 2021
  • Pacific herring, Clupea pallasii, a keystone species with significant ecological and commercial importance, is declining globally throughout much of its range. While traditional fishing equipment methods remain limited, new sensitive and rapid detection methods should be developed to monitor fisheries resources. To monitor the presence and quantity of C. pallasii from environmental DNA (eDNA) extracted from seawater samples, a pair of primers and a TaqMan® probe specific to this fish based on mitochondrial cytochrome b (COB) sequences were designed for the real-time PCR (qPCR) assay. The combination of our molecular markers showed high specificity in the qPCR assay, which affirmed the success of presenting a positive signal only in the C. pallasii specimens. The markers also showed a high sensitivity for detecting C. pallasii genomic DNA in the range of 1 pg~100 ng rxn-1 and its DNA plasmid containing COB amplicon in the range of 1~100,000copies rxn-1, which produced linear standard calibration curves (r2=0.99). We performed a qPCR assay for environmental water samples obtained from 29 sampling stations in the southeastern coastal regions of South Korea using molecular markers. The assay successfully detected the C. pallasii eDNA from 14 stations (48.2%), with the highest mean concentration in Jinhae Bay with a value of 76.09±18.39 pg L-1 (246.20±58.58 copies L-1). Our preliminary application of molecular monitoring of C. pallasii will provide essential information for efficient ecological control and management of this valuable fisheries resource.

Effects of The Different Dietary Arginine Levels on Plasma Free Amino Acid Concentrations in rainbow trout

  • Ok, I.H.;Bai, S.C.;Hung, S.S.O.;Rogers, Q.R.
    • Proceedings of the Korean Society of Fisheries Technology Conference
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    • 2001.05a
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    • pp.322-323
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    • 2001
  • Studies in the chick (Hill and Olsen 1963, Richardson et al.1953), rat (Swendseid et al. 1963, Young and Zamora 1968), pig (Puchal et al. 1962), man (Longnecker and Hause 1961, Snyderman et al. 1964, Swendseid et al. 1966), and fish (Thebault 1985) have clearly established that a reduced concentration of an EAA in plasma reflects a deficient level of that amino acid in the diet. (omitted)

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Evaluation of Sodium Intake and Relationship between Sodium Intake and the Bone Mineral Density of Female University Students (중부 지역 여대생에서 음식섭취빈도조사지를 이용한 나트륨 섭취량 평가 및 나트륨 섭취와 골밀도와의 관련성 조사)

  • Bae, Yun-Jung;Yeon, Jee-Young
    • Journal of the East Asian Society of Dietary Life
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    • v.21 no.5
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    • pp.625-636
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    • 2011
  • The purpose of this study was to evaluate the relationship between bone health and sodium intake in female university students using a dish frequency questionnaire (DFQ 125), anthropometric checkups, food records for 3 days, and ultrasound measurement of calcaneus bone mineral density. Subjects were divided into two groups: normal (n=196) and osteopenia (n=52). There were no significant differences in age or height between the two groups. The average weight, body mass index, and body fat in the normal group were significantly higher than in the osteopenia group. The sodium intake of DFQ was positively correlated with the sodium intake of 3 days of dietary records (p=0.0003). There were no significant differences in the sodium intake between the two groups from DFQ. The dishes were ranked by sodium intake: kimchies were 17.68%, noodles and mandu were 16.36%, stews were 13.69%, main dishes such as meat, egg, and beans were 11.47%, and fish and shellfish were 11.07%. The frequency of eating noodles and mandu (p=0.0116), stews (p=0.0008), kimchies (p=0.0482), fish and shellfish (p=0.0362), vegetables (p=0.0064) and seasoning (p=0.0347) were negatively associated with bone mineral density. Bone health was not significantly different with increasing quartiles of sodium intake. As excessive sodium intakes may indirectly affect bone mineral density, these results suggest that to prevent osteoporosis, university students needed to be more educated about diets containing less sodium through nutrition education programs.

Cattle Age Prediction by Leukocytes Telomere Quantification (혈액세포의 텔로미어 함량을 이용한 소의 연령예측)

  • Choi, Na-Eun;Kim, Hyun-Sub;Choe, Chang-Yong;Jeon, Gwang-Joo;Sohn, Sea-Hwan
    • Journal of Animal Science and Technology
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    • v.52 no.5
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    • pp.367-374
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    • 2010
  • Telomeres at the end of chromosomes consist of tandem repeats of (TTAGGG)n DNA sequence and associated proteins. Telomeres have the essential functions in chromosome stability and genome integrity and are hence related to cell senescence and cancer. This study was carried out to quantify the amount of telomeric DNA and establish age prediction equations by using the quantity of telomeric DNA for cattle. Analysis of the telomere quantity of the lymphocytes was performed at different age, across breeds and between different sexes of cattle. We quantified the amount of telomeric DNA by the Q-FISH technique using the telomeric DNA probe in 460 cattle at age of 1~166 months in Korean Cattle and Holstein breeds. In results, we found that the amount of telomeric DNA decreased gradually with age. The amount of telomeric DNA of Korean Cattle was significantly higher than that of Holstein breed (P<0.01). In addition, the amount of telomeric DNA in male was significantly higher than that in female (P<0.01). Using the relationship between age and the amount of telomeric DNA in cattle, age predicting equations were established as a result of regression analysis. Because sex and breeds influenced telomeric DNA quantity, the age prediction equations were estimated separately in Korean Cattle females and Holstein females. The regression equations were $\hat{Y}$=$38.102X^2$-220.103X + 318.309 (P<0.0001, $R^2$=0.8019) in Korean Cattle females and $\hat{Y}$ = $42.799X^2$ - 199.682X + 242.106 (P<0.0001, $R^2$ = 0.8379) in Holstein females, where the X was quantity of telomeric DNA and Y was predicted age in months. These equations predicted the age of cattle with high significance and accuracy and have high R square values. Thus, it could be possible to scientifically predict the age using the above equations for Korean Cattle and Holstein females.

Effect of Maternal Age on Chromosome Aberrations and Telomere Quantity in Chick Embryos (닭의 모체 연령에 따른 생산 배아의 염색체 이상 빈도 및 텔로미어 함량 분석)

  • Lee, Soo-Hee;Subramani, Vinod K.;Sohn, Sea-Hwan
    • Korean Journal of Poultry Science
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    • v.36 no.4
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    • pp.293-300
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    • 2009
  • The rate of fetus with abnormal chromosomes increase with maternal age. Nondisjunction of aging oocyte chromosome is a major reason for the increased rate of abnormalities. Telomeres are the ends of eukaryotic chromosome, which are essential for chromosome stability and are related in cell senescence. This study was carried out to analyze the chromosome aberration rate and amount of telomeric DNA in chick embryo along with maternal age. Fertilized eggs and blood were sampled from White Leghorn layers starting at 20 weeks through to 70 weeks age at 10 weeks interval. Chromosome aberration rate was analyzed by karyotyping. The amounts of telomeric DNA in embryonic cells and lymphocytes were quantified by Quantitative Fluorescence in situ Hybridization method. The chromosome aberration rate in chick embryos significantly differed with maternal age. The chromosome aberration rate increased at early laying period and beyond 70 weeks of maternal age. Therefore, chromosome aberration rate was affected by maternal age due to ovulated oocytes state. However, the amount of telomeric DNA on embryonic cells did not differ significantly with maternal age. Thus, maternal age does not affects telomere quantity in their embryos due to cellular reprograming at early embryonic stage after fertilization.

Amount of Telomeric DNA on Lymphocytes in Senescence Mouse by Quantitative Fluorescence in situ Hybridization (노화촉진마우스의 텔로미어 함량 분석)

  • Lee, Mi-Rang;Do, Kyoung-Tag;Han, Jyung-Ju;Moon, So-Hyun;Kang, Han-Seok;Kim, Seon-Ku;Shin, Teak-Soon;Lee, Hong-Goo;Hwang, Dae-Yon;Kim, Yong-Gyun;Sohn, Sea-Hwan;Choi, Na-Eun;Kim, Byeong-Woo;Cho, Byung-Wook
    • Journal of Life Science
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    • v.19 no.10
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    • pp.1463-1467
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    • 2009
  • Telomeres, comprised of tandem repeats of TTAGGG sequences, are special nucleoprotein structures that protect and stabilize chromosome ends. These structures form the crux of the telomere concept of aging, senescence and genomic instability. The classic terminal restriction fragment (TRF) analysis to quantify the amount of telomeric DNA is disadvantageous in species containing ultra long telomeres like in mice (100Kb). In this study, we used a more sensitive quantitative fluorescence in situ hybridization (Q FISH) technique to quantify telomeric DNA, and used it as a biological aging marker in mice. 12 litters each of Senescence-Resistant (SAMR1) and -Prone (SAMP1) known as senescence accelerated mouse strains were purchased from Central Lab, Animal Inc. We quantified the amount of telomeric DNA using telomere specific DNA probes on the two strains of male mice at 8 weeks, 18 weeks and 26 weeks of age. The amount of telomeric DNA correlated with aging and age associated changes in body and organ weight between SAMR1 and SAMP1 strains of mice. These data suggest the usefulness of the amount of telomeric DNA as a biological aging marker in human aging studies.

KiSS1, KiSS2, GPR54 mRNA Expression of the Blacktip Grouper Epinephelus fasciatus (홍바리 Epinephelus fasciatus의 KiSS1, KiSS2, GPR54 mRNA 발현양상)

  • Kang, Hyeong-Cheol;Lee, Chi-Hoon;Song, Young-Bo;Baek, Hea-Ja;Kim, Hyung-Bae;Lee, Young-Don
    • Development and Reproduction
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    • v.16 no.2
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    • pp.121-128
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    • 2012
  • Kisspeptin has been reported to facilitate sexual maturation and ovulation by directly stimulating GnRH neurons via its receptor, GPR54. The KiSS-GPR54 system is playing an important role in the reproduction of several mammalian species. However, little is known about their function in fish. The aim of this study is to understand the physiological function and evolutionary conservation of KiSS-GPR54 system in teleost fish blacktip grouper, Epinephelus fasciatus. In the present study, we have partial cloned KiSS1, KiSS2 GPR54 mRNAs from a brain samples. Tissue distribution analysis using RT-PCR revealed that the KiSS1, KiSS2, GPR54 transcripts were expressed in different tissue. The KiSS-GPR54 system in gonadal of immature and mature stage were analyzed using qRT-PCR. The partial sequence of KiSS1, KiSS2, GPR54 were 232 bp, 304 bp, 613 bp long, respectively. KiSS1, KiSS2, GPR54 mRNAs are shown common expression in the brain. The amount of KiSS1, KiSS2 mRNAs expression were significantly higher in mature stage than immature stage. However GPR54 mRNA expression was higher in immature stage. These results are in good agreement with the hypothesis that KiSS-GPR54 system plays an important role in the regulation of reproductive function in the blacktip grouper.