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Isolation and Genetic Transformation of Primordial Germ Cell (PGC)-Derived Cells from Cattle, Goats, Rabbits and Rats

  • Lee, C.K.;Moore, K.;Scales, N.;Westhusin, M.;Newton, G.;Im, K.S.;Piedrahita, J.A.
    • Asian-Australasian Journal of Animal Sciences
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    • v.13 no.5
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    • pp.587-594
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    • 2000
  • At present embryonic stem (ES) cells with confirmed pluripotential properties are only available in the mouse. Recently, we were able to isolate, culture and genetically transform primordial germ cell (PGC)-derived cells from pig embryos and demonstrate their ability to contribute to chimera development in the pig. In order to determine whether the system we developed could be used to isolate embryonic germ (EG) cells from other mammalian species, we placed isolated PGCs from cattle, goats, rabbits and rats in culture. Briefly, PGCs were isolated from fetuses of cow (day 30-50), goat (day 25), rabbit (day 15-18) and rat (day 11-12), and plated on STO feeder cells in Dulbecco's modified Eagle's medium (DMEM): Ham's F10 medium (1:1) supplemented with 0.01 mM nonessential amino acids, 2 mM L-glutamine, 0.1 mM $\beta$ - mercaptoethnol, soluble recombinant human stem cell factor (SCF; 40ng/ml), human basic fibroblast growth factor (bFGF; 20ng/ml) and human leukemia inhibitory factor (LIF; 20ng/ml). For maintenance of the cells, colonies were passed to fresh feeders every 7-10 days. In all species tested, we were able to obtain and maintain colonies with ES-like morphology. Their developmental potential was tested by alkaline phosphatase (AP) staining and in vitro differentiation assay. For genetic transformation, cells were electroporated with a construct containing the green fluorescent protein (GFP) under the control of the cytomegalovirus (CMV) promoter. GFP-expressing colonies were detected in cattle, rabbits and rats. These results suggest that PGC-derived cells from cattle, goats, rabbits and rats can be isolated, cultured, and genetically transformed, and provide the basis for analyzing their developmental potential and their possible use for the precise genetic modification of these species.

Mineralogy and Geochmistry of the Sanjeon Au-Ag Deposit, Wonju Area, Korea (산전 금-은 광상에 관한 광물 및 지화학적 연구)

  • Se-Hyun Kim
    • Economic and Environmental Geology
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    • v.32 no.5
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    • pp.445-454
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    • 1999
  • The Sanjeon Au-Ag deposit consists of three subparallel hydrothermal quartz-calcite veins which filled fault-related fractures (generally $N20^{\circ}$ to 35"W-trending and $70^{\circ}$ to $80^{\circ}$ SW-dipping) within quartz porphyry. The vein mineralization shows an apparent variation of mineral assemblages with paragenetic time: (1) early, white quartz + pyrite + arsenopyrite + brown sphalerite, (2) middle, white (vein) to clear quartz (vug) + base-metal sulfides + electrum + argentite, (3) late, calcite + pyrite + native silver. Mineralogic and fluid inclusion data indicate that gold-silver minerals were deposited at temperatures from 2l $0^{\circ}$ to $250^{\circ}$ with salinities of 4 to 5 wt. % equiv. NaCl and log fS2 values from -14.0 to -12.2 atm. The linear relationship between homogenization temperature and salinity data indicates that gold-silver deposition was a result of meteoric water mixing. Ore mineralization occurred at pressure conditions of about 70 bars, which corresponds to the mineralization depths of about 260 m to 700 m. There is a remarkable decrease of the calculated 1)180 values of water from 1.3 to -9.7%0 in hydrothermal fluid with increasing paragenetic time. This indicates a progressive increase of meteoric water influx in the hydrothermal system at the Sanjeon deposit. Oxygen-hydrogen, sulfur, and carbon isotope values of hydrothermal fluids indicate that the ore mineralization was formed largely from meteoric waters with the contribution of sulfur and carbon from a deep igneous source.

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Hematological Change of Korean Native Goats Experimentally Infected with Bovine Leukemia Virus (Bovine Leukemia Virus에 실험감염된 한국재래산양의 혈액상의 변동)

  • 이필돈;김종호;전무형
    • Korean Journal of Veterinary Service
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    • v.18 no.1
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    • pp.1-21
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    • 1995
  • To elucidate pathogenesis of bovine leukemia virus(BLV) in Korean native goats, the goats experimentally infected with BLV were studied especially for the aspects of infectivity and hematological changes. The experimental goats were examined for 27 months by agar-gel immunodiffusion(AGID) test and syncytium formation assay. During this period, changes of total leucocyte, absolute Iymphocyte and atypical Iymphocyte were examined, and the distribution of surface immunoglobulin ( sIg ) -bearing cells and rosette forming cell (RFC) in the peripheral Iymphocyte were also investigated. By indirect immunofluorescence (IFA) and complement dependent antibody cytotoxicity (CDAC) assay using monoclonal antibody(Mab) against bovine leukosis tumor-associated anti-gen(BL-TAA), changes of BL-TAA positive Iymphocyte in peripheral blood were measured. The results obtained through the experiment were summarized as follows. 1. Antibody titers were measured by AGID using gP51 and P24 antigens. The animals were serologically converted at 2 months post-inoculation(pi) in gP51 antigen, whereas sero-converted at 4 months pi in P24 antigen. In comparison with antibody titers for gP51, P24 antigen showed lower titers throughout the trial period. 2. The peripheral lymphocytes from all of the infected goats, as co-cultivated with F8l cells manifested syncytial formation at 4 months pi. 3. On counting total leucocyte, Iymphocyte and atypical Iymphocyte, two out of four infected goats showed normal distribution, while No 2 of the remaining two revealed temporal and No 3, Persistant increasing number of the cells. 4. The optimal condition of rosette formation of the peripheral Iymphocyte of normal Korean native goats was shown in the sheep erythrocyte treated with 0.1M AET for 30 nun at $37^{\circ}C$. When the Iymphocytes were treated in nylon wool column, the number of sIg-bear-ing cell were increased in the nylon wool adherent cells, but RFC was increased in the non-adherent cells. Of the infected goats, No 2 and No 3 showed significantly increasing number of sIg-bearing cells at 18 months pi. 5. The Iymphocytes of No 2 and No 3 goats reacted positively in IFA using Mab against BL-TAA at 12 months pi and 18 months pi, respectively. In CDAC test, all of four infected goats revealed positive reaction at 24 months pi. The higher positive rates were observed in No 2 and No 3 as compared with the remainders.

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The Fatigue Crack Growth Behavior of Concrete (콘크리트의 피로균열 성장거동에 관한 연구)

  • 김진근;김윤용
    • Magazine of the Korea Concrete Institute
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    • v.9 no.3
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    • pp.127-135
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    • 1997
  • In this study, the wedge splitting tcst (WST) was carried out for the fatigue wack growth behavior of concrete. Selected test variables were concrete compressive strength of 28, 60 and 118 MI%, and stress ratio with 2 levels (6. 13%). In oder to make the designed stress ratio, the maximum and thr minimum fatigue loading level were 75-85% and 5- 10% of ultimate static load, respectively. Fatigue testing was preceded by crack mout.h opening displacement (CMOI)) compliance calibration tcst, and then the fatigue crack growth was computed by crack lcngth vs. (lMOI) compliance relations acquisited by the CMOD compliance calibration technique. To evaluate thc validity of CMOD compliancc calibration techniquc, the crack length p~mlicted by this method was cornpard with the crack length by linear elastic fracture mechanics(LEFIbl) and dyeing test. On the basis of the experimental results, a LRFhl-based c.mpirica1 model for f'at,igue crack growth rate(da/dN-AKI relationships) was presented. The fat,igut. crack growth ratc increased with the strength of concwtc. It appcars that t.he da/tiN-AKI relationships was influenced by stress ratio, however, the effect is diminished with an increase of strength. The comparisons between CblOl) compliance calibration technique anti the other. methods gave the validity of' ('MOD compliance calibration technique for the LZXT.

Mineral Metabolism in Singleton and Twin-pregnant Dairy Goats

  • Harter, C.J.;Castagnino, D.S.;Rivera, A.R.;Lima, L.D.;Silva, H.G.O.;Mendonca, A.N.;Bonfim, G.F.;Liesegang, A.;St-Pierre, N.;Teixeira, I.A.M.A.
    • Asian-Australasian Journal of Animal Sciences
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    • v.28 no.1
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    • pp.37-49
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    • 2015
  • During pregnancy, the maternal body undergoes significant physiological changes. The present study assessed the changes on calcium (Ca), phosphorus (P), magnesium (Mg), sodium (Na) and potassium (K) metabolism in singleton and twin-pregnant dairy goats. The 42 goats used ($49.5kg{\pm}7.6$ body weight [BW]) were assigned at random to treatments that were factorially arranged to account for 2 breeds (Oberhasli and Saanen), 2 pregnancy types (singleton and twin) and 3 gestation periods (80, 110, and 140 days). Digestibility trials were performed at 80, 110, and 140 days of gestation. Mineral retention during pregnancy was determined in the maternal body, femur, uterus, mammary gland, fetus and fetal fluid. Blood samples were taken during pregnancy before and after a meal, and Ca, P, Mg, Na, K ions and alkaline phosphatase activity determined in serum. Bone mineral density was determined in the right femur. Statistical analyses were performed using the SAS MIXED procedure. Dry matter intake decreased linearly up to 140 days of gestation. Maternal BW gain, and Ca, P, and Mg retention (g/kg) decreased linearly with the advance of gestation days. Macromineral retention in maternal body (g/kg) was greater in Oberhasli than Saanen goats, and their fetuses had higher Ca, P, and Mg deposition (mg/g). Mineral retention (mg/g) increased in fetuses according to pregnancy development, with no differences between singleton and twin pregnancy. In the mammary gland, the retention of all minerals (g) increased with the days of pregnancy. In conclusion, related to Ca, P, and Mg metabolism can be divided into two stages. Up to 80 days of gestation, was characterized by the preparation of the maternal body reserves for future mineral demands. From 80 days of gestation onward, was characterized by the transfer of maternal body reserves for fetal development and colostrum production. Na and K supply was provided by adjustments in endogenous excretion and an increase in intestinal absorption. Finally, mineral metabolism was specific to each genotype and, except for Na, was not affected by the number of fetuses.

Saccharomyces cerevisiae Live Cells Decreased In vitro Methane Production in Intestinal Content of Pigs

  • Gong, Y.L.;Liao, X.D.;Liang, J.B.;Jahromi, M.F.;Wang, H.;Cao, Z.;Wu, Y.B.
    • Asian-Australasian Journal of Animal Sciences
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    • v.26 no.6
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    • pp.856-863
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    • 2013
  • An in vitro gas production technique was used in this study to elucidate the effect of two strains of active live yeast on methane ($CH_4$) production in the large intestinal content of pigs to provide an insight to whether active live yeast could suppress $CH_4$ production in the hindgut of pigs. Treatments used in this study include blank (no substrate and no live yeast cells), control (no live yeast cells) and yeast (YST) supplementation groups (supplemented with live yeast cells, YST1 or YST2). The yeast cultures contained $1.8{\times}10^{10}$ cells per g, which were added at the rates of 0.2 mg and 0.4 mg per ml of the fermented inoculum. Large intestinal contents were collected from 2 Duroc${\times}$Landrace${\times}$Yorkshire pigs, mixed with a phosphate buffer (1:2), and incubated anaerobically at $39^{\circ}C$ for 24 h using 500 mg substrate (dry matter (DM) basis). Total gas and $CH_4$ production decreased (p<0.05) with supplementation of yeast. The methane production reduction potential (MRP) was calculated by assuming net methane concentration for the control as 100%. The MRP of yeast 2 was more than 25%. Compared with the control group, in vitro DM digestibility (IVDMD) and total volatile fatty acids (VFA) concentration increased (p<0.05) in 0.4 mg/ml YST1 and 0.2 mg/ml YST2 supplementation groups. Proportion of propionate, butyrate and valerate increased (p<0.05), but that of acetate decreased (p<0.05), which led to a decreased (p<0.05) acetate: propionate (A: P) ratio in the both YST2 treatments and the 0.4 mg/ml YST 1 supplementation groups. Hydrogen recovery decreased (p<0.05) with yeast supplementation. Quantity of methanogenic archaea per milliliter of inoculum decreased (p<0.05) with yeast supplementation after 24 h of incubation. Our results suggest that live yeast cells suppressed in vitro $CH_4$ production when inoculated into the large intestinal contents of pigs and shifted the fermentation pattern to favor propionate production together with an increased population of acetogenic bacteria, both of which serve as a competitive pathway for the available H2 resulting in the reduction of methanogenic archaea.

Population Dynamics of Mabled sole Limanda yokohamae($G{\"{U}}NTHER$) in Tokyo Bay, Japan (동경만산 문치가자미Limanda yokohamae($G{\"{U}}NTHER$)의 자원양 변동의 해석)

  • PARK Jong-Soo;SIMIZU Mako-to
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.24 no.1
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    • pp.1-8
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    • 1991
  • Population dynamics of Matted sole Limanda yokohamae($G{\"{U}}NTHER$) in Tokyo bay, Japan has been studied by virtual population analysis (VPA) for multi cohort and experimental fishing. Based on the biological data, the present parameters of the Limanda yekohamae stock at the Tokyo bay, Japan were estimated as follows: natural mortality coefficient(M) were 0.313 for male and 0.250 for female, terminal fishing mortality coefficient(F) were 2.190 for male, and 0.798 for female, rate of exploitation(E) was $30\%\;to\;50\%$. From the result of virtual population analysis for multi cohort, the population size were estimated from 3,5000,000 to 9,200,000 fishes, according to the result of experimental fishing, estimated stock size were 2,400,000 to 8,700,000 fishes. Stock size difference of the two methods were about two times in 1987, however, other years has been showed from 0.8 to 1.5 times. Both method has been showed same increase and decrease tendency of the c. p. u. e. and catches. From the isopleth diagram plot by Beverton and Holt's yield per recruit, the catches could be increase two times for female, 1.3 times for male than present aspects by the fishing management. And further, as reducing fishing effort, extension of mesh size and rising the length at first caputre, are reasonable in order to manage the stock at the optimum level.

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Effect of Long-Term Pyridoxine Depletion on Asparte Aminotransferase and Pyridoxal 5'-Phosphate of Rat Liver Mitochondrial and Cytosolic Fractions (장기간의 Pyridoxine 부족이 쥐 간의 Mitochondria 및 Cytosolic Fraction 에 있는 Aspartate Aminotransferase 및 Pyridoxal 5'- Phoshate에 미치는 영향)

  • 임경숙
    • Journal of Nutrition and Health
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    • v.19 no.4
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    • pp.246-254
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    • 1986
  • Weanling female Sprague Dawley rats were fed d diets containing 22mg pyridoxine. BCI/kg diet (control diet) and l.2mg pyridoxine. BCI/kg diet (deficient diet). One control group and one defi­c dent group were fed their diet throughout growth, g gestation and lactation. After the pups were born and weaned, the deficient group was divided into two groups. One switched to control diet(supple­I mented group) and the other continued the same d deficient diet( deficient group) until 10 week -old. The liver mitochondrial and cytosolic asparate a aminotransferase activity and pyridoxal phosphate content were determined in offspring rats. The aspartate aminotransferase activities in both liver mito$\phi$ondrial and cytosolic fractions of den­d cient group were significantly lower than those of controls, but there were no significant differences between two groups after addition of 1O^{-4}M pyri­d do뼈I phosphate to the medium. By pyridoxine s supplementation after weaning, the reduced aspar­a tate aminotnmsferase activities were only partialy I restored to control levels. The pyridoxal phospha­t te content of deficient group in Iiver mitochondr­ial and cytosoIic fractions were alo significantly different from those of controls, but readily restored by dietary supplementation. These results suggest that there is a quantitative and a qualitative changes of aspartate amino trans­f ferase and pyridoxal phosphate in liver mitochon­d drial and cytosolic fraction by long-term pyrido­x xine deficiency and these reductions can partially recovered by dietary pyridoxine supplementation after weaning.

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Effects of Dietary Energy Levels on the Physiological Parameters and Reproductive Performance of Gestating Gilts

  • Jin, S.S.;Jung, S.W.;Jang, J.C.;Chung, W.L.;Jeong, J.H.;Kim, Y.Y.
    • Asian-Australasian Journal of Animal Sciences
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    • v.29 no.7
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    • pp.1004-1012
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    • 2016
  • This experiment was conducted to investigate the effects of dietary energy levels on the physiological parameters and reproductive performance of gestating first parity sows. A total of 52 F1 gilts ($Yorkshire{\times}Landrace$) were allocated to 4 dietary treatments using a completely randomized design. Each treatment contained diets with 3,100, 3,200, 3,300, or 3,400 kcal of metabolizable energy (ME)/kg, and the daily energy intake of the gestating gilts in each treatment were 6,200, 6,400, 6,600, and 6,800 kcal of ME, respectively. During gestation, the body weight (p = 0.04) and weight gain (p = 0.01) of gilts linearly increased with increasing dietary energy levels. Backfat thickness was not affected at d110 of gestation by dietary treatments, but increased linearly (p = 0.05) from breeding to d 110 of gestation. There were no significant differences on the litter size or litter birth weight. During lactation, the voluntary feed intake of sows tended to decrease when the dietary energy levels increased (p = 0.08). No difference was observed in backfat thickness of the sows within treatments; increasing energy levels linearly decreased the body weight of sows (p<0.05) at d 21 of lactation and body weight gain during lactation (p<0.01). No significant differences were observed in the chemical compositions of colostrum and milk. Therefore, these results indicated that high-energy diets influenced the bodyweight and backfat thickness of sows during gestation and lactation. NRC (2012) suggested that the energy requirement of the gestation gilt should be between 6,678 and 7,932 kcal of ME/d. Similarly, our results suggested that 3,100 kcal of ME/kg is not enough to maintain the reproductive performance for gilts during gestation with 2 kg feed daily. Gilts in the treatment 3,400 kcal of ME/kg have a higher weaning number of piglets, but bodyweight and backfat loss were higher than other treatments during lactation. But bodyweight and backfat loss were higher than other treatments during lactation. Consequently, an adequate energy requirement of gestating gilts is 6,400 kcal of ME/d.

Estimation of Genetic Parameters and Trends for Weaning-to-first Service Interval and Litter Traits in a Commercial Landrace-Large White Swine Population in Northern Thailand

  • Chansomboon, C.;Elzo, M.A.;Suwanasopee, T.;Koonawootrittriron, S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.23 no.5
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    • pp.543-555
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    • 2010
  • The objectives of this research were the estimation of genetic parameters and trends for weaning-to-first service interval (WSI), and litter traits in a commercial swine population composed of Landrace (L), Large White (T), LT, and TL animals in Chiang Mai, Northern Thailand. The dataset contained 4,399 records of WSI, number of piglets born alive (NBA), litter weight of live piglets at birth (LBW), number of piglets at weaning (NPW), and litter weight at weaning (LWW). Variance and covariance components were estimated with REML using 2-trait analyses. An animal model was used for WSI and a sire-dam model for litter traits. Fixed effects were farrowing year-season, breed group of sow, breed group of boar (litter traits), parity, heterosis (litter traits), sow age, and lactation length (NPW and LWW). Random effects were boar (litter traits), sow, permanent environment, and residual. Heritabilities for direct genetic effects were low for WSI (0.04${\pm}$0.02) and litter traits (0.05${\pm}$0.02 to 0.06${\pm}$0.02). Most heritabilities for maternal litter trait effects were 20% to 50% lower than their direct counterparts. Repeatability for WSI was similar to its heritability. Repeatabilities for litter traits ranged from 0.15${\pm}$0.02 to 0.18${\pm}$F0.02. Direct genetic, permanent environment, and phenotypic correlations between WSI and litter traits were near zero. Direct genetic correlations among litter traits ranged from 0.56${\pm}$0.20 to 0.95${\pm}$0.05, except for near zero estimates between NBA and LWW, and LBW and LWW. Maternal, permanent environment, and phenotypic correlations among litter traits had similar patterns of values to direct genetic correlations. Boar genetic trends were small and significant only for NBA (-0.015${\pm}$0.005 piglets/yr, p<0.004). Sow genetic trends were small, negative, and significant (-0.036${\pm}$0.013 d/yr, p<0.01 for WSI; -0.017${\pm}$0.005 piglets/yr, p<0.007, for NBA; -0.015${\pm}$0.005 kg/yr, p<0.01, for LBW; -0.019${\pm}$0.008 piglets/yr, p<0.02, for NPW; and -0.022${\pm}$0.006 kg/yr, p<0.003, for LWW). Permanent environmental correlations were small, negative, and significant only for WSI (-0.028${\pm}$0.011 d/yr, p<0.02). Environmental trends were positive and significant only for litter traits (p<0.01 to p<0.0003). Selection based on predicted genetic values rather than phenotypes could be advantageous in this population. A single trait analysis could be used for WSI and a multiple trait analysis could be implemented for litter traits.