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Hypomethylation of DNA in Nuclear Transfer Embryos from Porcine Embryonic Germ Cells

  • Lee, Bo-Hyung;Ahn, Kwang-Sung;Heo, Soon-Young;Shim, Ho-Sup
    • Journal of Embryo Transfer
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    • v.27 no.2
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    • pp.113-119
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    • 2012
  • Epigenetic modification including genome-wide DNA demethylation is essential for normal embryonic development. Insufficient demethylation of somatic cell genome may cause various anomalies and prenatal loss in the development of nuclear transfer embryos. Hence, the source of nuclear donor often affects later development of nuclear transfer (NT) embryos. In this study, appropriateness of porcine embryonic germ (EG) cells as karyoplasts for NT with respect to epigenetic modification was investigated. These cells follow methylation status of primordial germ cells from which they originated, so that they may contain less methylated genome than somatic cells. This may be advantageous to the development of NT embryos commonly known to be highly methylated. The rates of blastocyst development were similar among embryos from EG cell nuclear transfer (EGCNT), somatic cell nuclear transfer (SCNT), and intracytoplasmic sperm injection (ICSI) (16/62, 25.8% vs. 56/274, 20.4% vs. 16/74, 21.6%). Genomic DNA samples from EG cells (n=3), fetal fibroblasts (n=4) and blastocysts from EGCNT (n=8), SCNT (n=14) and ICSI (n=6) were isolated and treated with sodium bisulfite. The satellite region (GenBank Z75640) that involves nine selected CpG sites was amplified by PCR, and the rates of DNA methylation in each site were measured by pyrosequencing technique. The average methylation degrees of CpG sites in EG cells, fetal fibroblasts and blastocysts from EGCNT, SCNT and ICSI were 17.9, 37.7, 4.1, 9.8 and 8.9%, respectively. The genome of porcine EG cells were less methylated than that of somatic cells (p<0.05), and DNA demethylation occurred in embryos from both EGCNT (p<0.05) and SCNT (p<0.01). Interestingly, the degree of DNA methylation in EGCNT embryos was approximately one half of SCNT (p<0.01) and ICSI (p<0.05) embryos, while SCNT and ICSI embryos contained demethylated genome with similar degrees. The present study demonstrates that porcine EG cell nuclear transfer resulted in hypomethylation of DNA in cloned embryos yet leading normal preimplantation development. Further studies are needed to investigate whether such modification affects long-term survival of cloned embryos.

Effect of Monensin and Live Yeast Supplementation on Growth Performance, Nutrient Digestibility, Carcass Characteristics and Ruminal Fermentation Parameters in Lambs Fed Steam-flaked Corn-based Diets

  • Ding, J.;Zhou, Z.M.;Ren, L.P.;Meng, Q.X.
    • Asian-Australasian Journal of Animal Sciences
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    • v.21 no.4
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    • pp.547-554
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    • 2008
  • In performance, digestibility and slaughter trials, a total of forty five male weaned lambs were used to examine the effects of monensin and live yeast supplementations on growth performance, nutrient digestibility, carcass characteristics and ruminal fermentation parameters when the lambs were fed steam-flaked corn-based diets. Animals were allotted to one of three treatment diets in a completely randomized design. The three treatment diets were: (1) basal diet (CON) with steam-flaked corn as a sole grain source, (2) basal diet supplemented with monensin (MO), and (3) basal diet supplemented with live yeast (LY). Total average daily intake (ADI) was unaffected by MO and LY supplementations. LY supplementation increased (p<0.05) average daily gain (ADG) by 13.1% compared with the CON diet. Both MO and LY supplementations resulted in a significant improvement (p<0.05) of feed efficiency over the CON diet (4.47, 4.68 vs. 5.05). Hemicellulose digestibility was higher (p<0.05) for lambs in the LY supplementation group (62.4%) as compared with the CON group (55.7%), but no differences were observed in digestibilities of dry matter (DM), organic matter (OM), crude protein (CP), neutral detergent fiber (NDF) and acid detergent fiber (ADF). All carcass traits were not influenced by dietary supplementations. Ruminal pH in lambs fed the LY supplemental diet was more stable than that with the CON diet (6.57 vs. 6.17). Neither MO nor LY supplementation influenced the concentration of ruminal ammonia-N and total volatile fatty acid (VFA), and molar percentages of individual VFA. Plasma urea-N concentration was decreased (p<0.05) by MO and LY supplementations, while plasma ${\beta}$-hydroxybutyrate (BHBA), glucose and other blood parameters were unaffected. In conclusion, while both MO and LY supplementations had a positive impact on feed efficiency and LY supplementation stabilized ruminal pH and improved fiber utilization, none of the supplements had the capacity to significantly enhance the carcass characteristics.

Comparative Assay Fungus Population and Resistant Genes about Magnaporthe grisea between Korea and China (한국·중국 벼 도열병균 생리형 분석 및 품종 저항성 특성 비교)

  • Kim, Dong-Yun;Shim, Hong-Sik;Cailin, Lei;Ling, Zhong Zhuan;Han, Seong-Sook
    • Research in Plant Disease
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    • v.10 no.4
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    • pp.217-226
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    • 2004
  • One hundred isolates of Magnaporthe grisea from Korea and China were characterized for pathogenicity using eight Korean differential varieties(KDV), six Chinese differential varieties(CDV) and six near isogenic lines(NILs) developed in China. The restriction length polymorphism of M. grisea isolates from each country also was analyzed using MGR586 as a probe. One hundred Korean isolates classified into 17 races on KDV were grouped into 29 pathotypes on Chinese near isogenic lines(NILs). Virulence of 46% of Korean isolates against all the six Chinese NILs indicated that the current six Chinese NILs alone was not enough to be used as differential varieties in Korea. Especially, susceptibility of the BL1 carrying resistance gene Pi-b to 70% of tested Korean isolates suggested that BL1(Pi-b) may not be a useful resistance source to Korean blast. Based on the virulence assays of M. grisea populations from each country were divided into two groups. About 50% of Chinese isolates showed similarity to the 30% of the Korean isolates. Especially, the isolates from northern part of China, where Japonica rice cultivars were grown, showed high similarity to the Korean isolates, while isolates from southern part of China, where Indica rice were mainly grown, showed low similarity to Korean isolates. The genome RFLPs of Korean isolates were quite different from those of southern part of China using MGR586 as a probe. These data indicated that the physiological and genetical characteristics of M. grisea population might be determined by strong interaction with cultivated rice.

Feasibility Study of Parallel- Plate Detector Using Dielectric film for 6 MV X-ray (6MV X-선 검출특성 조사를 위하여 유전체 필름을 이용하여 제작한 평행판 검출기의 유용성)

  • 조문준;김용은;이병용;김정기;임상욱;김현수;김기환
    • Progress in Medical Physics
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    • v.15 no.2
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    • pp.105-111
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    • 2004
  • The parallel plate detector with dielectric film for dosimetry was designed to measure detection characteristic of 6 MV X-ray with medical linear accelerator. PTFE film was inserted into FEP films that are made by two one-side metal coated materials for ion source. The thicknesses of PTFE dielectric film was 100 ${\mu}{\textrm}{m}$ and the thickness of FEP dielectric film was 100 ${\mu}{\textrm}{m}$, respectively. This detector was fixed by two acrylic plate for physical hardness ad geometrical consistency. The geometrical condition for measurement with parallel-plate for detector was below; SSD=100 cm and the 5 cm depth between detector and phantom surface The major parameter of detector characteristics such as zero drift current, leakage current, charge response by applied voltage, reproducibility, linearity, TMR measurement, dose rate effect were measured. The zero drift currents are 8.3 pA and leakage currents are 10 pA. The charge response of applied voltage is showing linearity in 414 voltage. The measurement deviation of reproducibility in this detector is within 1% for dose and the linearity of applied dose shows in this detector. The TMR curves in phantom between this parallel plate detector and reference detector are matched within 3% deviation from maximum dose depth to 7.5 cm depth. It is considered that this dosimetric system is satisfactory for the purpose of the constancy check of the 6 MV x-ray from medical linear accelerator.

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Effects of Size and Rate of Maturing on Carcass Composition of Pasture- or Feedlot- Developed Steers

  • Brown, A.H. Jr.;Camfield, P.K.;Baublits, R.T.;Pohlman, F.W.;Johnson, Z.B.;Brown, C.J.;Tabler, G.T.;Sandelin, B.A.
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.5
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    • pp.661-671
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    • 2006
  • Steers (n = 335) of known genetic backgrounds from four fundamentally different growth types were subjected to two production systems to study the main effects and possible interactive effects on carcass composition. Growth types were animals with genetic potential for large mature weight (LL), intermediate mature weight-late maturing (IL), intermediate mature weight-early maturing (IE), and small mature weight-early maturing (SE). Each year, in a nine year study, calves of each growth type were weaned and five steers of each growth type were developed on pasture or feedlot and harvested at approximately 20 and 14 mo of age, respectively. Data recorded were chilled carcass weight and percentages of forequarter, foreshank, chuck, rib, plate, brisket, hindquarter, round, rump, shortloin, sirloin, flank, lean, fat, bone, and retail cuts. The growth $type{\times}production$ system interaction was an important source of variation in chilled carcass weight (p = 0.0395) and percentage retail cuts (p = 0.001), lean (p = 0.001), fat (p = 0.001), rump (p = 0.0454), shortloin (p = 0.0487), and flank (p = 0.001). The ranking of the growth $type{\times}production$ system means for percentage lean was LL-pasture>IL-pasture = IE-pasture = SE-pasture>LL-feedlot, IL-feedlot>IE-feedlot = SE-feedlot. The growth $type{\times}production$ system interaction was non-significant (p>0.05) for forequarter, foreshank, chuck, rib, plate, brisket, hindquarter, round and bone. Growth types of IE and SE yielded greater (p<0.05) mean forequarter than did growth types of IL and LL ($51.6{\pm}0.3$ and $51.5{\pm}0.3$ vs. $51.1{\pm}0.3$ and $50.8{\pm}0.3%$). Mean bone was highest (p<0.05) for the LL growth type and lowest (p<0.05) for the SE growth type ($19.5{\pm}0.5$ vs. $16.8{\pm}0.5%$). Mean bone was greater (p<0.05) for the pastured steers than for the feedlot steers ($21.8{\pm}0.8$ vs. $14.5{\pm}0.6%$). These data indicate that growth type responded differently in the two production systems and that these results should be helpful in the match of genetics to production resources.

Preparation and characterization of protein isolate from Yellowfin tuna Thunnus albacares roe by isoelectric solubilization/precipitation process

  • Lee, Hyun Ji;Lee, Gyoon-Woo;Yoon, In Seong;Park, Sung Hwan;Park, Sun Young;Kim, Jin-Soo;Heu, Min Soo
    • Fisheries and Aquatic Sciences
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    • v.19 no.3
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    • pp.14.1-14.10
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    • 2016
  • Isoelectric solubilization/precipitation (ISP) processing allows selective, pH-induced water solubility of proteins with concurrent separation of lipids and removal of materials not intended for human consumption such as bone, scales, skin, etc. Recovered proteins retain functional properties and nutritional value. Four roe protein isolates (RPIs) from yellowfin tuna roe were prepared under different solubilization and precipitation condition (pH 11/4.5, pH 11/5.5, pH 12/4.5 and pH 12/5.5). RPIs contained 2.3-5.0 % moisture, 79.1-87.8 % protein, 5.6-7. 4 % lipid and 3.0-3.8 % ash. Protein content of RPI-1 and RPI-2 precipitated at pH 4.5 and 5.5 after alkaline solubilization at pH 11, was higher than those of RPI-3 and RPI-4 after alkaline solubilization at pH 12 (P < 0.05). Lipid content (5.6-7.4 %) of RPIs was lower than that of freeze-dried concentrate (10.6 %). And leucine and lysine of RPIs were the most abundant amino acids (8.8-9.4 and 8.5-8.9 g/100 g protein, respectively). S, Na, P, K as minerals were the major elements in RPIs. SDS-PAGE of RPIs showed bands at 100, 45, 25 and 15 K. Moisture and protein contents of process water as a 2'nd byproduct were 98.9-99.0 and 1.3-1.8 %, respectively. Therefore, yellowfin tuna roe isolate could be a promising source of valuable nutrients for human food and animal feeds.

Antioxidative and Macrophage Phagocytic Activities and Functional Component Analyses of Selected Korean Chestnut (Castanea crenata S. et Z.) Cultivars (국내산 밤 일부 품종의 기능성 성분분석과 항산화 및 대식세포 활성)

  • Lee, Hyeon-Ju;Chung, Mi-Ja;Cho, Jae-Youl;Ham, Seung-Shi;Choe, Myeon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.37 no.9
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    • pp.1095-1100
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    • 2008
  • Antioxidative and macrophage phagocytic activities and contents of functional component in selected Korean chestnuts (Dantaek, Daebo, Okkwang, Seokchu, Byunggo) were evaluated. Coumarin, gallic acid and catechin in inner skin and whole kernel of selected Korean chestnuts were detected by HPLC. The predominant functional components in inner skin of chestnut were catechin, followed by gallic acid and coumarin. However, the whole kernel had only gallic acid. Thus, the antioxidant properties of gallic acid and catechin were evaluated through DPPH radical-scavenging activity and SOD like activity. Gallic acid and catechin at 6.0 mg/100 g exhibited 69.4% and 38.3% of scavenging activities on DPPH radical, respectively. DPPH radical-scavenging activity of gallic acid increased in a concentration-dependent manner. Gallic acid was found to possess higher DPPH radical-scavenging activity than equivalent amount of catechin at all concentrations, whereas catechin was found to have higher SOD like activity than gallic acid. In addition, pre-incubation of macrophage with white kernel extract from Byunggo resulted in a significant increase of phagocytic activity and yellow kernel extracts from Byunggo, Dantaek, Daebo and Okkwang, leading to an increase in phagocytic activity compared with untreated cells. Yellow kernel extracts was found to have higher phagocytic activity than white kernel extracts. Byunggo had the highest phagocytic activity. The results suggest that the Korean chestnut may provide a natural source of antioxidants and active immunity.

Determination and Prediction of the Amino Acid Digestibility of Sunflower Seed Meals in Growing Pigs

  • Liu, J.D.;Li, Q.Y.;Zeng, Z.K.;Li, P.;Xu, X.;Wang, H.L.;Zhang, S.;Piao, X.S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.28 no.1
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    • pp.86-94
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    • 2015
  • This experiment was conducted to evaluate the chemical composition and amino acid (AA) digestibility of sunflower seed meal (SFSM) and to use this data to develop prediction equations for estimating AA digestibility for growing pigs. Ten SFSM were collected from five provinces in China. Twelve barrows ($38.8{\pm}4.6kg$), fitted with ileal T-cannula were allotted into two $6{\times}6$ Latin square designs. Each of six experimental periods comprised a 5-d adaption period followed by a 2-d collection of ileal digesta. The ten test diets contained 50% SFSM as the sole source of AA. Another nitrogen-free diet was used to measure the basal endogenous losses of crude protein (CP) and AA. Chromic oxide (0.3%) was used as an inert marker in each diet. There was considerable variation (CV>10%) among the ten SFSM in chemical composition (dry matter [DM]). The concentration of CP and ether extract (EE) ranged from 29.33% to 39.09% and 0.88% to 11.33%, respectively. Crude fibre (CF), neutral detergent fibre and acid detergent fibre ranged from 21.46% to 36.42%, 38.15% to 55.40%, and 24.59% to 37.34%, respectively. There was variation among the ten SFSM in apparent ileal digestibility (AID) and standardized ileal digestibility (SID) for lysine and threonine, which ranged from 63.16 to 79.21 and 55.19% to 72.04% for AID and 67.03% to 82.07% and 61.97% to 77.01% for SID, respectively. The variation in CP and methionine ranged from 60.13% to 74.72% and 74.79% to 88.60% for AID and 66.70% to 79.31% and 77.16% to 90.27% for SID, respectively. Methionine was a good indicator to predict AA digestibility. These results indicate that conventional chemical composition of SFSM was variable (CV>10%) among the ten SFSM (DM). The results of AID, SID and prediction equations could be used to evaluate the digestibility of SFSM in growing pigs.

Nutritive Evaluation of Poultry Waste and Sudex Grass Silage for Sheep

  • Chaudhry, S.M.;Naseer, Z.;Alkraidees, M.S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.10 no.1
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    • pp.79-85
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    • 1997
  • Cage layer waste and sudex grass were ensiled in the proportions of 0:100, 30:70, 40:60 and 50:50 wet basis, respectively. The influence of ensiling cage layer waste on Salmonellae, Shigella, Proteus, and total number of colony forming units (CFU) was investigated. The nutritive value of the silages was evaluated in a digestion trial. The experiment was conducted with 24 wethers allotted to four silages. Initial samples of cage layer waste showed $0.11{\times}10^6$ CFU and salmonellae, Shigella and Proteus were present. Ensiling was effective in complete elimination of all the pathogens. Dry matter, crude protein and ash contents were increased linearly (p < .01) with the increase of cage layer waste in the silages ($358g{\cdot}kg^{-1}$ to $484g{\cdot}kg^{-1}$; $76.3g{\cdot}kg^{-1}$ to $183.2g{\cdot}kg^{-1}$ and $38.5g{\cdot}kg^{-1}$ to $169.4g{\cdot}kg^{-1}$; DM basis, respectively). Water solubles carbohydrate values for silages were 38.3, 22.5, 20.1 and $20.0g{\cdot}kg^{-1}$ DM basis, respectively. Ensiling decreased the pH values for all the silages and the decrease was higher for sudex grass ensiled alone than grass ensiled with cage layer waste. Lactic acid concentration in silages increased linearly (p < .01) with the increase of cage layer waste. The apparent digestibilities of DM, OM and CP for the animals fed sudex grass silage alone were 496.0, 516.1, $496.7g{\cdot}kg^{-1}$ DM basis respectively. However, digestibilities of all the components were higher (p < 0.01) for the animals fed silages containing cage layer waste. Among waste containing diets, the digestibilities of all the components showed quadratic affect (p < 0.01), increased for silages containing 30 and 40% cage layer waste and decreased when the level of waste increased from 40 to 50%. The results indicated that cage layer waste can be used upto 40% in ruminant diet as a source of N without any adverse effect on the health of animals. Ensiling appeared to be feasible and effective method for eliminating the pathogen present in cage layer waste.

Effects of Methionine Hydroxy Analogue on changes of Characteristics of food wastes in Incubator (메치오닌 수산화물의 첨가가 남은 음식물의 보관 중 성상변화에 미치는 영향)

  • Lee, H.J.;Kim, H.S.;Jeong, H.Y.;Kang, S.W.;Ki, K.S.;Cho, K.K.;Cho, J.S.;Hong, Z.S.;Jin, M.G.;Lee, H.G.;Choi, Y.J.
    • Journal of the Korea Organic Resources Recycling Association
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    • v.10 no.3
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    • pp.59-67
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    • 2002
  • This study was carried out to improve the utilization of food wastes known as major source of environmental pollution and to determine proper additional levels of methionine hydroxy analogue(MHA) on food wastes. 1%, 3% and 5% of MHA were added to 3 groups of food wastes gathered from small cafeteria. Each group has 3 reiteration. NaCl contents were tended to get higher as addition levels increased(p<0.05). And pH values were tended to get lower as additional levels increased(p<0.05). $NH_3-N$ contents were significantly decreased as additional levels increased(p<0.05). Total VFA(Volatile Fatty Acid) content was increased after 30 days of storage on control. On MHA treatments(1, 3, 5%), total VFA contents were decreased until 15 days of storage, and kept low values until the end of experiment. In terms of superficial change, mildew covered all over control, and covered by thin mildew on the surface of 1% MHA treatment. No superficial change found on 3% and 5% MHA treatments. In conclusion, over 1% addition of MHA was effective to prevent the decay and degeneration of food wastes.

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