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Cloning of Farm Animals in Japan; The Present and the Future

  • Shioya, Yasuo
    • Proceedings of the Korean Society of Developmental Biology Conference
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    • 2001.10a
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    • pp.37-43
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    • 2001
  • 1. About fifty thousand of cattle embryos were transferred and 16000 ET-calves were born in 1999. Eighty percents of embryos were collected from Japanese Black beef donors and transferred to dairy Holstein heifers and cows. Since 1985, we have achieved in bovine in vitro fertilization using immature oocytes collected from ovaries of slaughterhouse. Now over 8000 embryos fertilized by Japanese Black bull, as Kitaguni 7~8 or Mitsufuku, famousbulls as high marbling score of progeny tests were sold to dairy farmers and transferred to their dairy cattle every year. 2. Embryo splitting for identical twins is demonstrated an useful tool to supply a bull for semen collection and a steer for beef performance test. According to the data of Dr. Hashiyada(2001), 296 pairs of split-half embryos were transferred to recipients and 98 gave births of 112 calves (23 pairs of identical twins and 66 singletons). 3. A blastomere-nuclear-transferred cloned calf was born in 1990 by a joint research with Drs. Tsunoda, National Institute of Animal Industry (NIAI) and Ushijima, Chiba Prefectural Farm Animal Center. The fruits of this technology were applied to the production of a calf from a cell of long-term-cultured inner cell mass (1988, Itoh et al, ZEN-NOH Central Research Institute for Feed and Livestock) and a cloned calf from three-successive-cloning (1997, Tsunoda et al.). According to the survey of MAFF of Japan, over 500 calves were born until this year and a glaf of them were already brought to the market for beef. 4. After the report of "Dolly", in February 1997, the first somatic cell clone female calves were born in July 1998 as the fruits of the joint research organized by Dr. Tsunoda in Kinki University (Kato et al, 2000). The male calves were born in August and September 1998 by the collaboration with NIAI and Kagoshima Prefecture. Then 244 calves, four pigs and a kid of goat were now born in 36 institutes of Japan. 5. Somatic cell cloning in farm animal production will bring us as effective reproductive method of elite-dairy- cows, super-cows and excellent bulls. The effect of making copy farm animal is also related to the reservation of genetic resources and re-creation of a male bull from a castrated steer of excellent marbling beef. Cloning of genetically modified animals is most promising to making pig organs transplant to people and providing protein drugs in milk of pig, goat and cattle. 6. Farm animal cloning is one of the most dreamful technologies of 21th century. It is necessary to develop this technology more efficient and stable as realistic technology of the farm animal production. We are making researches related to the best condition of donor cells for high productivity of cloning, genetic analysis of cloned animals, growth and performance abilities of clone cattle and pathological and genetical analysis of high rates of abortion and stillbirth of clone calves (about 30% of periparutum mortality). 7. It is requested in the report of Ministry of Health, labor and Welfare to make clear that carbon-copy cattle(somatic cell clone cattle) are safe and heathy for a commercial market since the somatic cell cloning is a completely new technology. Fattened beef steers (well-proved normal growth) and milking cows(shown a good fertility) are now provided for the assessment of food safety.

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Effect of dietary mannanoligosaccharide supplementation on nutrient digestibility, hindgut fermentation, immune response and antioxidant indices in dogs

  • Pawar, Mahesh M.;Pattanaik, Ashok K.;Sinha, Dharmendra K.;Goswami, Tapas K.;Sharma, Kusumakar
    • Journal of Animal Science and Technology
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    • v.59 no.5
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    • pp.11.1-11.7
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    • 2017
  • Background: Use of prebiotics in companion animal nutrition is often considered advantageous over probiotics because of the ease of handling, ability to withstand processing and storage etc. While most of the studies on prebiotic use in dogs have been done with processed food as basal diet, the response in relation to homemade diet feeding is not very well explored. Methods: The study was conducted to evaluate the effects of dietary mannanoligosaccharide (MOS) supplementation on nutrient digestibility, hindgut fermentation, immune response and antioxidant indices in dogs. Ten Spitz pups were divided into two groups: control (CON) with no supplementation, and experimental (MOS) wherein the basal diet was supplemented with MOS at 15 g/kg diet. All dogs were fed on a home-prepared diet for a period of 150 days. The study protocol included a digestion trial, periodic blood collection and analysis for lipid profile and erythrocytic antioxidants. Immune response of the animals was assessed towards the end of the feeding period. Results: Results revealed no significant (P > 0.05) variations in palatability score, intake and apparent digestibility of nutrients between the groups. Faecal score, faeces voided, faecal pH, concentrations of ammonia, lactate and short-chain fatty acids were comparable (P > 0.05) between the two groups. Cell-mediated immune response, assessed as delayed-type of hypersensitivity response, was significantly higher (P < 0.05) in the MOS group. The percent of lymphocyte sub-populations CD4+ and ratio of CD4+:CD8+ were also significantly (P < 0.05) higher in MOS group. The serum IgG levels were similar (P > 0.05) in both the groups. Supplementation of MOS lowered (P < 0.05) serum total- and LDL- cholesterol levels, when compared with the control group. The erythrocytic antioxidant indices were similar (P > 0.05) between the two groups. Conclusions: The results indicated that supplementation of MOS at the rate of 15 g/kg in the diet of dog augmented the cell-mediated immune response and serum lipid profile without any influences on digestibility of nutrients, hindgut fermentation and antioxidants indices.

Effect of In Vivo Embryo Production and Embryo Transfer Following Superovulation in Hanwoo (다배란 처리에 따른 한우 체내 수정란 생산 효율과 수정란이식)

  • Cho, Sang-Rae;Choi, Sun-Ho;Choe, Chang-Yong;Son, Jun-Kyu;Choi, Soo-Ho;Kim, Young-Joo;Lee, Poong-Yeon;Yeon, Sung-Heum;Kim, Hyun-Jong;Son, Dong-Soo
    • Journal of Embryo Transfer
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    • v.25 no.3
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    • pp.141-144
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    • 2010
  • In vivo embryo produced from Hanwoo donor cows were collected and transferred to Hanwoo recipients. Cows, at random stages of the estrous cycle, received Progesterone Releasing Intravaginal Device (CIDR-plus, InterAg, New Zealand) together with injection of 1 mg estradiol benzoate and 50 mg progesterone, and gonadotropin treatment began 4 day later. For superovulation, a total of 28 mg FSH was intramuscularly injected twice a day in the way of decreasing doses 4 day (5, 5, 4, 4, 3, 3, 2 and 2 mg). Twenty one Hanwoo donor cows were flushed on day 7 of estrus cycle with same FSH and artificial insemination by the same technicians. Embryos were recovered 7 days after the second insemination by flushing the uterus with Embryo Collection Medium. The results obtained were as follows: The rates of transferable embryos were 50.3%, and 78 fresh embryos at morulae and blastocysts stage were transferred into Hanwoo recipients on day 7 of estrus cycle. The pregnancy rates were first embryo transfer 55.6%, 2nd 62.9% and 3rd 57.9%, respectively. In conclusion, These results suggest that CIDR-based superovulation protocol may be effectively used for production of superior Hanwoo embryos. Also, since it seems the condition of recipient cows greatly affect pregnancy rate, it is very important to evaluate recipient for effective cattle production.

Urine Specific Gravity as a Useful Tool for Screening Proteinuria in Children (소아 단백뇨 검사에 있어서 요비중의 유용성)

  • Kim, Jong-Hwa;Yoo, Kee-Hwan;Hong, Young-Sook;Lee, Joo-Won;Kim, Soon-Kyum
    • Childhood Kidney Diseases
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    • v.4 no.1
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    • pp.1-5
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    • 2000
  • Purpose: The plcr of spot urine has been uised to predict the timed urine protein excretion. Although this method reduces errors caused by variations in urine volume, it is relatively thconvenient and expensive. Recently, a more rapid and less expensive screening method with specific gravity(SG) has been reported, and we have examined whether estimated-creatinine(Cr-est) with urine 5G could be used in place of urine creatinine to predict 24-hour collected urine protein excretion in children. Methods: We had retrospectively analyzed protein, creatinine and urine SG in randomized spot urine samples of 147 patients from March 1998 till June 1998 in Korea university Guro hospital and compared the urinary protein creatinine ratio(P/Cr) with the protein estimated-creatinine ratio(P/Cr-est). We compared the correlation of urinary creatinine vs-urine 5G with the timed urine pretein excretion. Results: 1) urine SG accurately estimated urine creatinine concentration (r=0.407, P<0.001, Cr=SG x 4485.82-4482.87). 2) P/Cr correlated with urine protein excretion measured in a 24-hour urine collection (r=0.771, P<0.001, 24-hour collected urine protein : 0.338 x (P/Cr) 4+667.885). 3) P/Cr-est correlated with a 24-hour collected urine protein (r=0.723, P<0.001, 24-hour collected urine protein =0.354 x (P/Cr-est)+726.044), Conclusions: These results suggest that P/Cr-est with urine SG could be useful method for screening proteinuria in children.

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A study of Improvement on the Road Drainage Poor Site (도로배수 취약구간의 개선방안에 대한 연구)

  • Lee, Man-Seok;Kim, Heung-Rae;Lee, Kyung-Ha;Kang, Min-Soo;Song, Min-Tae
    • International Journal of Highway Engineering
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    • v.13 no.2
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    • pp.125-131
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    • 2011
  • This research aims to investigate the cause of the occurrence of a weak road drainage section scientifically and specifically through a site survey for a poorly drained section occurring due to rainfalls during road operation. This paper deeply reviewed the existing research results and current situation data on the poorly drained sections accumulated in Korea Expressway Corporation in order to investigate the cause of the occurrence of a weak road drainage section, and deeply verified and analyzed the weak sections for the road surface drainage facilities and the other road drainage facilities by visiting the expressway controlled by the 6 local headquarters and 33 branches of Korea Expressway Corporation. As a result of site surveys for the weak road drainage sections, i) in a road surface section, occurrence of ponding in the road shoulder pavement due to slope changes, bad collection of water in the collecting well at a median strip, shortage of road shoulder dike height, and inferior construction, etc. was analyzed to be the main cause of the occurrence of poorly drained sections, and ii) in a road neighborhood section, the occurrence of pavement height difference in a main road and shoulder section due to inferior ditches on a slope and the bad drain age at the inlet and outlet of a culvert due to soil deposits, debris, etc. were analyzed to be the main cause of the occurrence of weak sections. Proposed as a plan to improve the poorly drainage section of road were i)calculation of capacity through material changes at the ditch, enhancement of vertical sections and hydraulic analysis in terms of construction and other aspects, ii)derivation of a combined slope considering a slope and a vertical linearity and maintenance of proper distance between drainage structures in a vertical concave section in terms of geometrical structure, and iii)calculation of the drainage facility installation interval using a minutely rainfall intensity formula and a non-uniform flow analysis technique in terms of hydraulics and hydrologics and prompt removal of rainfalls from the road surface according to a linear drainage method.

Component proteins in cystic fluid of Taenia sodium metacestodes collected surgically from neurocysticercosis patients (인체 유구낭미충증 환자에서 수거한 낭미충 낭액의 성분 단백질의 양상)

  • Yoon Kong;Shin-Yong Kang;Seung-Yull Cho
    • Parasites, Hosts and Diseases
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    • v.28 no.2
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    • pp.101-108
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    • 1990
  • Surgically collected cystic fluid of Taenia solium metacestodes from patients of intracranial cystic lesion were compared in their protein composition with those from naturally infected pigs in Cheju Do, Korea and Ecuador. In non-denaturing discontinuous-polyacryla aide gel electrophoresis (disc-PAGE) , no discernible differences were recognized in banding patterns between the cystic fluids from Cheju Do and Ecuador, and between the cystic quids from pigs and human lesions except wider bands that corresponded to human albumin and T-globulin (in 4 of 9 patients). In reducing SDS-PAGE, bands in the cystic Ruid from Ecuador showed the same banding pattern with that from Cheju Do but two bands of 21 and 17 kDa were stained darker. Cystic quids (rom patients revealed the same protein compositions of the major protein bands of 94, 64, 15, 10 and 7 kDa as in the cystic fluid of pig origin, but human albumin (66 kDa), heavy and light chains of gamma globulin (55 and 22.5 kDa) were contaminated in 4 of 9 cystic fluids. Human CSF proteins seem to have been contaminated during cystic ftuid collection. In any cystic quid from patients, the majcr Protein component was 150 kDa which was subdivided into 15, 10 and 7 kDa in reducing SDS-PAGE.

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Genetic analyses of Acanthamoeba isolates from contact lens storage cases of students in Seoul, Korea

  • Yu, Hak-Sun;Choi, Kyung-Hee;Kim, Hyo-Kyung;Kong, Hyun-Hee;Chung, Dong-Il
    • Parasites, Hosts and Diseases
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    • v.39 no.2
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    • pp.161-170
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    • 2001
  • We conducted both the small subunit ribosomal DNA (SSU rDNA) polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and mitochondrial (mt) DNA RFLP analyses for a genetic characterization of Acanthamoeba isolates from contact lens storage cases of students in Seoul, Korea. Twenty-three strains of Acanthamoeba from the American Type Culture Collection and twelve clinical isolates from Korean patients were used as reference strains. Thirty-nine isolates from contact lens storage cases were classified into seven types (KA/LS1 , KA/LS2, KA/LS4, KA/LS5, KA/LS7 KA/LS18, KA/LS31). Four types (KA/LS1 , KA/LS2, KA/LS5, KA/LS18) including 33 isolates were regarded as A. castellanii complex by riboprints. KA/LS1 type was the most predominant (51.3%) in the present survey area, followed by KA/LS2 (20.9%), and KA/LSS (7.7%) types. Amoebae of KA/LS1 type had the same mtDNA RFLP and riboprint patterns as KA/E2 and KA/E12 strains, clinical isolates from Korean keratitis patients. Amoebae of KA/LS2 type had the identical mtDNA RFLP patterns with A. castellanii Ma strain, a corneal isolate from an American patient as amoebae of KA/LS5 type, with KA/E3 and KA/E8 strains from other Korean keratitis patients. Amoebae of KA/LS 18 type had identical patterns with JAC/E1, an ocular isolate from a Japanese patient. Three types , which remain unidentified at species level, were not corresponded with any clinical isolate in their mtDNA RFLP and riboprint patterns. Out of 39 isolates analyzed in this study, mtDNA RFLP and riboprint patterns of 33 isolates (84.6%) were identical to already known clinical isolates, and therefore, they may be regarded as potentially keratopathogenic. These results suggest that contact lens wearers in Seoul should pay more attention to hygienic maintenance of contact lens storage cases for the prevention of Acanthamoeba keratitis.

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Effect of Sphingosine-1-Phosphate on Intracellular Free Ca2+ in Cat Esophageal Smooth Muscle Cells

  • Lee, Dong Kyu;Min, Young Sil;Yoo, Seong Su;Shim, Hyun Sub;Park, Sun Young;Sohn, Uy Dong
    • Biomolecules & Therapeutics
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    • v.26 no.6
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    • pp.546-552
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    • 2018
  • A comprehensive collection of proteins senses local changes in intracellular $Ca^{2+}$ concentrations ($[Ca^{2+}]_i$) and transduces these signals into responses to agonists. In the present study, we examined the effect of sphingosine-1-phosphate (S1P) on modulation of intracellular $Ca^{2+}$ concentrations in cat esophageal smooth muscle cells. To measure $[Ca^{2+}]_i$ levels in cat esophageal smooth muscle cells, we used a fluorescence microscopy with the Fura-2 loading method. S1P produced a concentration-dependent increase in $[Ca^{2+}]_i$ in the cells. Pretreatment with EGTA, an extracellular $Ca^{2+}$ chelator, decreased the S1P-induced increase in $[Ca^{2+}]_i$, and an L-type $Ca^{2+}$-channel blocker, nimodipine, decreased the effect of S1P. This indicates that $Ca^{2+}$ influx may be required for muscle contraction by S1P. When stimulated with thapsigargin, an intracellular calcium chelator, or 2-Aminoethoxydiphenyl borate (2-APB), an $InsP_3$ receptor blocker, the S1P-evoked increase in $[Ca^{2+}]_i$ was significantly decreased. Treatment with pertussis toxin (PTX), an inhibitor of $G_i$-protein, suppressed the increase in $[Ca^{2+}]_i$ evoked by S1P. These results suggest that the S1P-induced increase in $[Ca^{2+}]_i$ in cat esophageal smooth muscle cells occurs upon the activation of phospholipase C and subsequent release of $Ca^{2+}$ from the $InsP_3$-sensitive $Ca^{2+}$ pool in the sarcoplasmic reticulum. These results suggest that S1P utilized extracellular $Ca^{2+}$ via the L type $Ca^{2+}$ channel, which was dependent on activation of the $S1P_4$ receptor coupled to PTX-sensitive $G_i$ protein, via phospholipase C-mediated $Ca^{2+}$ release from the $InsP_3$-sensitive $Ca^{2+}$ pool in cat esophageal smooth muscle cells.

Cancer Incidence in Kangwha County(1986-1992) (강화지역 암의 발생률(1986-1992))

  • Kim, Soh-Yoon;Kang, Hyung-Gon;Kim, Suk-Il;Yi, Sang-Wook;Ohrr, Hee-Choul
    • Journal of Preventive Medicine and Public Health
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    • v.32 no.4
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    • pp.482-490
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    • 1999
  • Objectives : This paper presents the information on the incidence of cancer from the population-based cancer resistry in Kangwha County. Material and methods : This investigation is based on Kangwha cancer registry. The data included cases of cancer diagnosed from 1986 through 1992. The diagnosis of cancer was confirmed by a team of physicians and nurses with the medical records kept in the clinics and hospitals based on the diagnostic criteria recommended by WHO. Home visitings were also made to cancer patients confirmed in every 6 months for the follow up and for the collection of relevant information directly from the patients. Results : A total of 992 cancer cases were registered during 1986-1992. The age-adjusted cancer incidence rate of all site is 201.7 in men and 110.7 in women. The most common cancer is the stomach cancer in both sexes. The age-adjusted incidence rate of the stomach cancer is 65.9 in men and 25.0 in women per 100,000 population. The lung cancer(33.8) and liver cancer(27.7) are next common cancers in men. The cervical cancer(21.8) and lung cancer(8.4) are next in women. Conclusion : The most common cancer is the stomach cancer in both sexes. The annual age-adjusted incidence rate of the stomach cancer is 65.9 in men and 25.0 in women per 100,000 population.

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A Case Study on Teachers' Teaching Professionalism for Secondary Science-Gifted Students (중등 과학영재 지도교사의 수업 전문성에 관한 사례연구)

  • Pae, Mi-Jung;Kim, Heui-Baik
    • Journal of The Korean Association For Science Education
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    • v.30 no.4
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    • pp.412-428
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    • 2010
  • The purpose of this study is to identify the teaching professionalism of the two teachers for secondary science-gifted students in perspective of teaching orientations. Both teachers have been taught biology to secondary science-gifted students for more than six years and they have received in-service training in gifted education. Teachers' orientations were investigated through in-depth interviews and observing lessons. For the data collection, videotapes were recorded during two lessons and two in-depth interviews for each participant were conducted. All recorded data were carefully transcribed and analyzed. Some unique characteristics of teaching were identified from the class instruction of each participant. Both teachers revealed 'Open Inquiry' orientation. This was reflected by their educational goals and beliefs that they should help science-gifted students to grow themselves as scientists by enhancing their inquisitiveness and creative problem solving ability. However, each teacher had shown different teaching orientations such as 'Academic Rigor' and 'Discovery' that seemed to have influence on the level or the range of subject matter covered in the classes. Teachers' science teaching orientations have been changed by their subject of teaching and their experiences in teaching the gifted. In the process, teachers' educational philosophy about gifted education plays an important role in teaching orientation. Based on the ongoing teachers' efforts for enhancing his/her professionalism, the teachers seemed to critically review theoretical knowledge of the science teachers in gifted education through self-reflection on their own teaching methods. When teacher's educational philosophy about gifted education was established and internalized, science teaching orientations in teaching practice seemed to be consistent with his/her goal of teaching.