• Title/Summary/Keyword: Transgenic animal

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Development of Reversing the Usual Order of Somatic Cell Nuclear Transfer in Mice

  • Kang, Ho-In;Sung, Ji-Hye;Roh, Sang-Ho
    • Journal of Embryo Transfer
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    • v.26 no.1
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    • pp.85-89
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    • 2011
  • Somatic cell nuclear transfer (SCNT) is a useful tool for reproducing genetically identical animals or producing transgenic animals. Many reports have demonstrated that the efficiency of animal cloning by SCNT requires reprogramming of the somatic nucleus to a totipotent like-state. The SCNT-related reprogramming might mimic the natural reprogramming process that occurs during normal mammalian development. However, recent evidence indicates that the reprogramming event by SCNT is incomplete. In this study, the traditional SCNT procedure (TNT) was modified by injecting donor nuclei into recipient cytoplasm prior to the enucleation process to expose the donor nucleus before removing the karyoplast containing the chromosomes of the oocytes which might possess additional reprogramming factors, and this modified technique was named as reversing the usual order of SCNT (RONT). Other procedures including activation and in vitro culture were the same as TNT. Contrary to expectations, the rate of blastocyst development was not different significantly between RONT and TNT (8.6% and 7.9%, respectively). However, duration of micromanipulation performed by the same technician and equipments was remarkably reduced because the ruptured oocytes after nuclear injection were excluded from the enucleation process. This study suggests that RONT, a simplified SCNT protocol, shortens the duration of SCNT procedure and this less time-costing protocol may enable the researchers to perform murine SCNT easier.

The Efficacy of Ozone Ointment Therapy on Pododermatitis of Dairy Cows (젖소 족피부염(足皮膚炎)에 대한 오존연고의 치료(治療) 효과(效果))

  • Lee, Soo-Jin;Jun, Moo-Hyung;Cho, Sung-Whan;Kim, Duck-Hwan;Park, Chang-Sik;Kim, Myung-Cheol
    • Journal of Veterinary Clinics
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    • v.23 no.3
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    • pp.279-285
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    • 2006
  • Thirty dairy cows with pododermatitis were selected and treatment effect of ozone ointment for bovine pododermatitis was investigated. In addition, bactericidal effect of ozone ointment on etiological agent of bovine pododermatitis was evaluated. The pathohistological examination for the pododermatitis, according to application with ozone ointment was investigated. Thirty dairy cows were divided two groups: control group(vaseline group: 15 cows), treatment group(ozone ointment group: 15 cows). Various parameters were evaluated in terms of the lameness score, swelling score, lesion score, WBC, neutrophil, pathohistological finding, and antimicrobial action. As compared with vaseline group, ozone ointment group revealed significant decrease of lameness(p<0.05), swelling(p<0.01) and lesion score(p<0.05) were shown in hoof lesions on 14 days after application. In hematological findings, WBC count revealed slightly high values within normal range before treatment, however, this was 1Corresponding author improved on 14 days after application of ozone ointment. The number of neutrophils was slightly higher than that of normal, however, this was improved on 14 days after application of ozone ointment. In pathohistological findings, normal dermal tissue was found in tissues with pododermatitis on 14 days after application of ozone ointment. In antimicobial action, marked decrease rate of bacteria was observed in feet of all cases treated with ozone ointment. The decreasing rate of bacteria in anaerobic culture was higher than that in aerobic culture.

The Efficacy of Ozonated Water Therapy on Pododermatitis of Dairy Cows (젖소 족피부염(足皮膚炎)에 대한 오존수의 치료(治療) 효과(效果))

  • Lee, Soo-Jin;Cho, Sung-Whan;Jun, Moo-Hyung;Kim, Duck-Hwan;Park, Chang-Sik;Han, Hong-Ryul;Kim, Myung-Cheol
    • Journal of Veterinary Clinics
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    • v.23 no.3
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    • pp.272-278
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    • 2006
  • This study was carried out to determine therapeutic the effect of ozonated water therapy on bovine pododermatitis. In addition, bactericidal effect of ozonated water on etiological agent of bovine pododermatitis was examined. The pathohistological examination for the pododermatitis, according to application with ozonated water and ozone ointment was investigated. Thirty healthy cattle were divided two groups(each of 15) : control group(povidone group), treatment group(ozone solution group). Various parameters were evaluated in terms of the lameness score, swelling score, lesion score, WBC count, neutrophil count, pathohistological finding, and antimicrobial action. The decrease of lameness and lesion score were shown in hoof lesions on 14 days after application of ozonated water. Significant decrease of swelling was shown in hoof lesions on 14 days 1Corresponding author after application of ozonated water(p<0.01). In hematological findings, WBC count revealed values within normal range. The number of neutrophils was slightly higher than that of normal, however, this was improved on 14 days after application of ozonated water. In pathohistological findings, recovery was rapid macroscopically and microscopically in the treatment with ozonated water on the hoof lesions and ozonated water was effective. In antimicobial action, bactericidal effect was observed in treatment with ozonated water on the hoof lesions and ozonated water was effective.

Antioxidant Effects of Ascorbic Acid on Renal-Ischemia Reperfusion Injury in Rabbit Model

  • Kim, Jong-Man;Lee, Jae-Yeon;Kim, Duck-Hwan;Jeong, Seong-Mok;Park, Chang-Sik;Kim, Myung-Cheol
    • Journal of Veterinary Clinics
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    • v.25 no.3
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    • pp.165-169
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    • 2008
  • Renal ischemia-reperfusion (I/R) injury is great clinical important because viability of the organ depends on the tolerance to ischemia-reperfusion injury, an inevitable processing during surgery. The purpose of this study was to investigate the effects of premedicated ascorbic acid alone in I/R injury model induced by cross-clamping of renal vessels. In the rabbit models, 2-4 kg New Zealand white rabbits were subjected to 30 minutes of warm unilateral renal ischemia followed by removal of contralateral kidney and then divided into five groups, control (2) arid treatment groups (3). In control group 1, the rabbits only received right nephrectomy. In control group 2, the rabbits received I/R on left kidney after the right nephrectomy. In treatment group 1, the rabbits received ascorbic acid 50 mg/kg IV before the operation. In treatment group 2, the rabbits received ascorbic acid 100 mg/kg IV before the operation. In treatment group 3, the rabbits received ascorbic acid 200 mg/kg IV before the operation. Blood samples were collected from these rabbits for measurement of kidney function tests at the 0, 1 st, 3rd and 7th day and antioxidant enzyme( SOD, GSHPx, CAT) at 24 hours. Kidney function tests (serum creatinine and BUN) showed a significant difference between group 2 and group 4, 5. Activity of antioxidant enzymes in plasma were significant decrease in group 4, 5 compare to group 2. The result of this study suggested that the exogenous ascorbic acid had a role of attenuation of renal I/R injury in rabbit model.

Study on germline transmission by transplantation of spermatogonial stem cells in chicken

  • Lee, Young-Mok;Han, Jae-Yong
    • Proceedings of the Korea Society of Poultry Science Conference
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    • 2006.11a
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    • pp.43-58
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    • 2006
  • As a bioreactor, bird has proved to be most efficient system for producing useful therapeutic proteins. More than half of the egg white protein content derives from the ovalbumin gene with four other proteins(lysozyme, ovomucoid, ovomucin and conalbumin) present at levels of 50 milligrams or greater. And the naturally sterile egg also contains egg white protein at high concentration allowing for a long shelf life of recombinant protein without loss in activity. In spite of these advantages, transgenic procedures for the bird have lagged far behind because of its complex process of fertilized egg and developmental differences. Recently, a system to transplant mouse testis cells from a fertile donor male to the seminiferous tubules of an infertile recipient male has been developed. Spermatogenesis is generated from transplanted cells, and recipients are capable of transmitting the donor haplotype to progeny. After transplantation, primitive donor spermatogonia migrate to the basement membrane of recipient seminiferous tubules and begin proliferating. Eventually, these cells establish stable colonies with a characteristic appearance, which expands and produces differentiating germ cells, including mature spermatozoa. Thus, the transplanted cells self-renew and produce progeny that differentiate into fully functional spermatozoa. In this study, to develop an alternative system of germline chimera production that operates via the testes rather than through developing embryos, the spermatogonial stem cell techniques were applied. This system consisted of isolation and in vitro-culture of chicken testicular cells, transfer of in vitro-maintained cells into heterologous testes, production of germline chimeras and confirmation of germline transmission for evaluating production of heterologous, functional spermatozoa.

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Plant-based production of therapeutic antibodies (식물기반 치료용 항체생산)

  • Kim, Young-Kwan;So, Yang-Kang;Park, Da-Young;Kim, Hyun-Soon;Jeon, Jae-Heung;Choo, Young-Kug;Ko, Ki-Sung
    • Journal of Plant Biotechnology
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    • v.37 no.3
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    • pp.262-268
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    • 2010
  • Antibodies are powerful and versatile tools to play a critical role in the diagnosis and treatment of many diseases. Their application has been enhanced significantly with the advanced recombinant DNA and heterologonous expression technologies, allowing to produce immunotherapeutic proteins with improved biofunctional properties. However, with currently available technologies, mammalian cell-based therapeutic antibody production, as an alternative for production in humans and animals, is often not plentiful for passive immunotherapeutics in treatment of many diseases. Recently, plant expression systems for therapeutic antibodies have become well-established. Thus, plants have been considered to provide an attractive alternative production system for therapeutic antibodies, as plants have several advantages such as the lack of human pathogens, and low cost of upstream production and flexible scale-up of highly valuable recombinant glycoproteins. Recent advances in modification of posttranslational processing for human-like glycosylation in transgenic plants will make it possible that plant can become a suitable protein expression system over the animal cellbased current production system. This review will discuss recent advances in plant expression technology and issues for their application to therapeutic antibody production.

Development of Detection Method of Unapproved Genetically Modified Potato (EH92-527-1) in Korea using Duplex Polymerase Chain Reaction (Duplex PCR을 이용한 국내 미승인 유전자변형 감자(EH92-527-1)의 검사법 개발)

  • Yoo, Myung-Ryul;Kim, Jae-Hwan;Yea, Mi-Chi;Kim, Hae-Yeong
    • Korean Journal of Food Science and Technology
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    • v.45 no.2
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    • pp.156-160
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    • 2013
  • A duplex polymerase chain reaction (PCR) method was developed to detect unapproved genetically modified (GM) potato (EH92-527-1) in Korea. The UDP-glucose pyrophosphorylase (UGP) gene was selected as an endogenous reference gene for potato and used to validate the specificity for 14 different crops. The primer pair EH92-F/R was designed to amplify the junction sequence between the genome and transgenic region introduced in GM potato. Its specificity was also validated using several different GM events. The detection limit of the duplex PCR method is approximately 0.05%. This duplex PCR method could be useful for monitoring cultivation of unauthorized GM potato in Korea.

Comparative study of wound healing in canine stomach with CO2 laser and scalpel incisions (CO2 레이저와 scalpel을 이용한 위 절개 시 개에서의 창상 치유 평가)

  • Lee, Jae-Yeon;Hwang, You-Sun;Kim, Han-Kyul;Choi, Hyun-Suk;Jeong, Seong-Mok;Cho, Sung-Whan;Park, Chang-Sik;Kim, Myung-Cheol
    • Korean Journal of Veterinary Research
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    • v.50 no.2
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    • pp.155-160
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    • 2010
  • The objective of this study was to compare wound healing in stomach with $CO_{2}$ laser and scalpel incision by measuring the extent of bleeding, the ease of gastric incision, incision time, degree of adhesion and wound healing degree in dogs. Sixteen healthy dogs were used. Two symmetrical incisions were made in ventral aspect of the stomach between the greater and lesser curvatures were made with scalpel and 0.2 mm spot diameter $CO_{2}$ laser (8W, continuous wave) in sixteen dogs. And then each wound was closed with absorbable suture in a two-layer inverting seromuscular pattern. At 3, 7, 14 and 21 days after initial wounding, each wound was taken for histological observation. On surgery, the extent of bleeding, the ease of incision and incision time showed significant differences between the groups. The $CO_{2}$ laser provided better hemostasis (p < 0.05) and smaller postoperative adhesion compared with the scalpel. However, the scalpel produced faster speed of incision and was easier to handle than the $CO_{2}$ laser (p < 0.05). Although there was no considerable difference between the two groups in histological observation, necrosis and calcium deposit tended to be larger in the $CO_{2}$ laser than in the scalpel.

Nonspecific Mouse Hepatitis Virus Positivity of Genetically Engineered Mice Determined by ELISA

  • Han, Dae Jong;Kim, Hyuncheol;Yeom, Su-Cheong
    • Biomedical Science Letters
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    • v.21 no.1
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    • pp.9-14
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    • 2015
  • Mouse hepatitis virus (MHV) is a major pathogen in laboratory mice that usually leads to fatal diseases, such as hepatitis, multiple sclerosis, encephalitis, and respiratory disease. MHV has a high infection rate, and it needs to be detected as soon as possible to prevent its spread to other facilities. However, MHV detection by enzyme-linked immunosorbent assay (ELISA) often gives false positives; thus, it is very important that the results are confirmed as true positives in the early infection stage or distinguished as false positives with more accurate, reliable methods. Under microbiological screening, MHV ELISA-positive mice were found in four GFP-tagging transgenic mice. To verify the detection of the MHV antigen directly, reverse transcription polymerase chain reaction (RT-PCR) was performed, and the mice were determined to be MHV negative. Additional serum antibody-based screening was conducted with three different ELISA kits, and multiplexed fluorometric immunoassay (MFIA) was performed to confirm their accuracy/sensitivity. In brief, the ELISA kit for A59 nucleocapsid protein (MHV-A59N) revealed MHV ELISA positivity, while other ELISA kits (MHV-S lysate and MHV-JHM lysate) demonstrated MHV negativity. In MFIA, only the test for the recombinant A59 nucleocapsid antigen was MHV positive, which was consistent with the ELISA results. These results suggest that the ELISA kit with the recombinant A59 nucleocapsid antigen might induce non-specific MHV ELISA positivity and that confirmation is therefore essential.

Histological Observation of Canine Acupoints (개에서 경혈의 조직학적 관찰)

  • Kim Myung-Cheol;Nam Tchi-Chou;Kim Moo-Kang;Kim Jong-Man;Kim Duck-Hwan;Lee Kyoung-Youl;Song Chi-Won;Park Chang-Sik
    • Journal of Veterinary Clinics
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    • v.23 no.2
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    • pp.102-104
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    • 2006
  • The purpose of this study was to document the histology of canine acupoints. Acupunctural needles were inserted by 0.5 to 1 cm depth into acupoints Nei-guan (Inner pass, PC06), Gan-shu (Liver Association Point, BL18), Shen-shu (Kidney Association Point, BL23) and Pangguang-shu (Bladder Association Point, BL28) in 4 dogs, and the tissues around the acupoints were examined. Light microscopy was used to observe the surrounding structures of each point. Nerve fibres, small vessels and muscle spindles were found around the tip of the needle in every case, although they occurred not so often at nonacupoint in nearby region or nonacupoint in the areas, such as skin, subcutaneous tissue and muscle. Therefore, it is suggested that nerve fibres, small vessels and muscle spindles may be potential acupoint receptors.