• 대한수의학회지
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• 제45권4호
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• pp.545-552
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• 2005

The major protocol features of the rodent uterotrophic assay have been evaluated using a range of reference chemicals. The protocol variables considered include the selection of the test species and route of chemical administration, the age of the test animals, the maintenance diet used, and the specificity of the assay for estrogens. The rodents were ovariectomized under general anesthesia via bilateral flank incisions and randomly assigned to groups of 5 animals. Chemicals were DEHP, DBP, BPA and NP, were injected sc once daily with combinations of chemicals treatments for 3 days. In the results, the reported estrogenic chemicals DEHP and DBP were both negative in the single dose treatments. But, in the combinations of chemicals treatments, DEHP and DBP increased in bud number of mammary gland. Treatment of ovariectomized mice with combinations of other chemicals resulted in uterine and vaginal hyperplasia. The additive estrogenic effects were seen with the combinations of $17{\beta}$-Bestradiol and DBP treatment. the competitive estrogenic effects were seen with the combinations of $17{\beta}$-Bestradiol and nonylphenol, $17{\beta}$-Bestradiol and bisphenol-A treatments. These results offers a sysmatic and mechanistically informative approach to assessing estrogenicity. it provides a useful profile of activity using a reasonable amount of resources and is compatible with the study of individual chemicals as well as the investigation of interactions among combinations of chemicals. The results described illustrate the intrinsic complexity of evaluating chemicals for estrogenic activities and conform the need for rigorous attention to experimental design and criteria for assessing estrogenic activity.

• Reproductive and Developmental Biology
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• 제29권4호
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• pp.235-239
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• 2005

This study was conducted to test whether the transgenic cattle pass the transgene to their progeny through germ cells, and whether the transgene is expressed in the mammary gland of ransgenic cows. Two male ransgenic calves were born from IVF-derived embryos injected with bovine $\beta-casein/human$ lactoferrin fusion gene and then grew up to be reproducible. Semen was collected from a transgenic bull after 18 mon of age and then frozen. Bovine oocytes matured in vitro were fertilized with spermatozoa of the transgenic bull and cultured in $50\;{\mu}L$ drops of CRlaa medium supplemented with 3 mg/mL BSA. After 48 h of culture, cleaved embryos were determined for the presence of transgenes by DNA polymerase chain reaction (PCR). Proportion of transgene positives among bovine embryos fertilized with sperm of the transgenic bull was $20.9\%$ (28/134). One of transgenic bulls did not produce transgenic sperm. Out of 34 calves produced from recipient heifers inseminated with semen of the other bull, 3 $(8.8\%)$ were transgenic animals (2 females and 1 male). Thus, one transgenic bull showed a low transmission frequency below Mendelian levels in both the IVF-derived embryos and his progeny. It was demonstrated by Southern blot that copy numbers of the transgene in the transgenic progeny enhanced about 1.8 times as compared to those of the founder bull The results demonstrate that the transgenic bull carrying human lactoferrin gene could pass his transgene to the progeny through germ cells, although he is a germ-line mosaic.

• 약학회지
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• 제47권2호
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• pp.57-68
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• 2003

Under the active search for biologically active novel agents for cancer prevention and treatment, some agents have been found from plants which are easily available. Our previous research on them revealed that C. annuum L. var. angulosum Mill have high antiproliferating effect on cancer cells. However, it has not been known whether the anticancer efficacy is different according to each part of C. annuum L. var. angulosum Mill or whether it can be changed by timing of harvest or solvent for extraction. Thus we compared the efficacy of each part of C. annuum L. var. angulosum Mill and assessed how much difference in the efficacy can be made according to the time of harvest or solvents for extraction. We observed the morphologic change and apoptosis 48 hr after treatment with the extract of each part of C. annuum L. var. angulosum Mill in MCF-7 mammary gland adenocarcinoma cells and human hepatoma cells. We also counted cancer cells by trypan blue method and MTT method to check the cytotoxicity. The leaf extract showed the highest anticancer effect among all the parts of C. annuum L. var. angulosum Mill; 50％ and 70％ reduction in the number of cancer cells was observed at 25 $\mu\textrm{g}$/mι and 50 $\mu\textrm{g}$/mι, respectively. It was more than 2 times as potent as 5-fluorouracil (5-FU). We found chromosomal fragmentation, clumping, and destuction by PI staining, and DNA fragmentation by electrophoresis. In conclusion, this study suggests that leaf extraction using water as solvent has the highest antiproliferative and apoptotic activity in cancer cells compared with other parts of extraction.

• Journal of Animal Science and Technology
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• 제61권2호
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• pp.61-68
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• 2019

The hG-CSF (human Granulocyte Colony-Stimulating Factor) is a growth and stimulation factor capable of inducing the proliferation of bone marrow cells, several types of leukocytes, among other hematopoietic tissue cells. hG-CSF is used in used to treat anomalies that reder a small number of circulating white blood cells, which may compromise the immune defenses of the affected person. For these reasons, the production of hG-CSF in a bioreactor system using the mammary gland of genetic modified animals is a possibility of adding value to the bovine genetic material and reducing the costs of hG-CSF production in pharmaceutical industry. In this study, we aimed the production of transgenic hG-CSF bovine through the lipofection of bovine primary fibroblasts with an hG-CSF expression cassette and cloning these fibroblasts by the somatic cell nuclear transfer (SCNT) technique. The bovine fibroblasts transfected with the hG-CSF cassette presented a stable insertion of this construct into their genome and were efficiently synchronized to G0/G1 cell cycle stage. The transgenic fibroblasts were cloned by SCNT and produced 103 transferred embryos and 2 pregnancies, one of which reached 7 months of gestation.

• 한국임상수의학회지
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• 제38권6호
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• pp.285-289
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• 2021

An eight-year-old, 3.4 kg, spayed female poodle dog was referred to Gyeongsang National University Animal Medical Center with obstinate bilateral retroperitoneal subcutaneous abscess. The medical history revealed ovariohysterectomy (OHE), lipoma, and mammary gland tumor excision performed two years ago. The dog presented with left retroperitoneal subcutaneous abscess discharge five months back and was treated by abscess drainage and antibiotics in a local veterinary hospital, but a relapse was reported on discontinuing antibiotics. On physical examination, the dog was febrile (40℃) and both retroperitoneal masses showed typical inflammatory reaction and purulent discharge. On serum chemistry analysis, hyperproteinemia, hyperglobulinemia, and elevated levels of C-reactive protein were observed. Cytologic examination showed degenerative neutrophils and phagocytic macrophages. Radiological examination revealed encapsulated hypoechoic subcutaneous masses in bilateral abdominal flank and the left flank mass was connected with the caudal pole of left kidney through fistula. Based on the diagnostic examination, exploratory celiotomy was recommended and left nephrectomy, and resection of bilateral masses were performed. During procedure, fistula between caudal pole of left kidney and left subcutaneous abscess was detected and suture material was identified. Resected tissue was histologically examined and diagnosed as suppurative abscess caused by the suture material. This case report describes chronic suppurative foreign body reaction including caudal pole of left kidney and bilateral retroperitoneal subcutaneous masses induced by the suture material used in OHE two years ago and the necessity of computed tomography examination to identify character of mass and extent of surgical resection.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2001년도 춘계학술발표대회
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• pp.60-60
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• 2001

Human thrombopoietin (hTPO) is a cytokine that plays a central role in megakaryopoiesis by influencing on the development and maturation of megakaryocyte and platelet production. To induce hTPO production in the mammary gland, expression vector was constructed by combining the promoter of bovine beta-casein gene, cDNA of hTPO and neomycine resistance gene for transfection into fibroblasts. Bovine fibroblast cells derived from female ear skin were transfercted with the expression vector using Lipofectamine (Life Technology, NY). Transected cells resistant to G4l8 treatment (600 $\mu\textrm{g}$/$m\ell$) were recovered and colony formation was initiated at 13 days. The colonies with about 1 cm diameter were picked and analysed by PCR. Single transfected cells were individually transferred to enucleated oocytes. After electrofusion, the reconstructed embryos were exposed to calcium ionophore (5uM) for 5 min followed by treatment with 6-DMAP (2.5 mM) for 4h. The nuclear transfer embryos were cultured in CRlaa medium at 38.5C, 5% $CO_2$ for 7 days. Twenty three of 29 (79.3%) colonies were proved to be hTPO transfectants by PCR. The colonies were further passaged and used to produce transgenic embryos using nuclear transfer. Cleavage and developmental rates of reconstructed embryos to the blastocyst stage were 65.1% and 39.4%, respectively Of 22 blastocysts that developed from reconstructed embryos with the transfected cell, 20 embryos (90.9%) were positive for hTPO by using PCR analysis. The results suggest that somatic cell nuclear transfer is efficient for production of transgenic embryos.

• 대한수의학회지
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• 제63권4호
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• pp.34.1-34.9
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• 2023

Dogs exhibit patterns of health issues that vary by life stage. An understanding of disease incidence with respect to breed and age/life stage could be an important component of canine health management and welfare. This study aimed to describe the age-specific disease incidence of 3 small dog breeds that attended veterinary clinics in the Republic of Korea, based on data from electronic veterinary medical records (EVMRs). A total of 40,785 EVMRs from Maltese (n = 21,355), Miniature Poodle (n = 11,658) and Shih Tzu dogs (n = 7,772) were analyzed. Common health problems in 3 small dog breeds were 'diseases of the skin' and 'diseases of the ear,' respectively. Among dogs aged ≤ 3 years, 'preventive medicine' was the most common cause cited for veterinary clinic visits. Among dogs aged 4 to 8 years, the most frequent health problems were 'diseases of the skin.' Among dogs aged 9 to 13 years, 'heart diseases,' 'kidney diseases,' 'mammary gland tumor,' and 'neoplasia (unspecified)' were considerably more frequent, compared to the rates in dogs ≤3 years. Among dogs aged ≥ 14 years, 'heart diseases' and 'sneezing/cough' were the main health problems. In all breeds, the frequencies of 'diseases of circulatory system,' 'diseases of respiratory system,' 'diseases of the nervous system,' 'endocrine' and 'neoplasia' increased rapidly with aging. This surveillance could inform strategies for disease screening tests and management based on life stage in these dog breeds and enable more effective health management.

• Journal of Dairy Science and Biotechnology
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• 제16권2호
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• pp.119-136
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• 1998

젖은 모든 새로 태어난 동물의 많은 필수영양소와 에너지를 공급해 주며, 유제품의 가공에 영향을 주는 독특한 물리적 특성을 지니고 있다. 본 연구에서는 동물의 유지방에 존재하는 주요 혹은 미량 성분의 조성과 구조를 비교하고, 갖 태어난 어린 동물의 영양요구량과 관련된 연구결과를 검토하고자 한다. 젖의 내용물과 조성은 동물의 종류에 따라 매우 다양하며, 이는 다양한 환경에서 적응하기 위한 진화과정에 의한 것으로 알려져 있다. 동물에 따라 젖의 지방종의 분포는 유사하지만 지방산의 조성은 일반적으로 매우 복잡하고 특이하다. 이는 각종 동물의 식이에서 오는 지방산과 유선에서 생합성된 지방산의 성격이 서로 다르기 때문이다. 특징적으로 젖의 지방산은 다른 조직에서는 발견되지 않는 짧은 사슬지방산과 중간 사슬지방산을 함유하고 있으나, 대부분의 동물에서 젖의 주요 지방인 중성지방은 구조적으로 매우 유사한 경향을 보이고 있다. 우유는 어린 동물에 많은 필수영양소와 에너지를 공급해준다. 이러한 영양소들은 우유가공 중 많은 물리적 특성에 영향을 준다. 우유의 주요물질과 미량물질을 비교함으로써 신생동물에 영양요구량을 예견할 수도 있다. 우유의 조성분은 동물에 따라 상당히 다르며 그것은 주로 환경에 적응하기 위해서 진화된 과정 중에 생겨난 것이다. 우유의 지방종 분포는 비슷하지만 지방산의 조성은 일반적으로 매우 복잡하고 특성이 있다. 이들 지방산 유선에서 합성되거나 사료섭취에 의한 것이다. 특징적으로 우유에서 발견되는 짧은 지방산과 긴 사슬의 지방산은 다른 조직에서는 잘 발견되지 않는다. 우유에 주요 지방인 중성지질은 동물의 종류에 따라서 매우 다양하지만 구조는 비슷하다. 실제로 우유 중의 지방구 형성과정은 잘 알려져 있지 않으나 세포의 ER로 부터 유례된 TG를 함유한 소포체가 지방구의 핵을 이룬다는 지적이 있다. 최근에는 각종 혈관질환관련 질병의 증가로 우유의 지방산을 해양동물에서 온 EPA나 DHA와 같은 불포화지방산으로 대체하고자 하는 연구가 많이 이루어지고 있다.

• 한국식품위생안전성학회지
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• 제13권2호
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• pp.106-111
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• 1998

본 연구는 bisphenol A의 실제 식품을 통한 사람의 노출농도에서 에스트로젠 효과를 알아보기 위하여 사람의 유방암 세포인 MCF-7와 난소를 적출한 무흉선 마우스를 이용하여 살펴보았다. MCF-7세포를 이용한 genistein 과 bisphenol A가 용매대조군과 무처치대조군에 비하여 genistein(10nM, 100nM, 1uM, 10uM)과 bisphenol A(2ng/ml, 4ng/ml, 8ng/ml, 16ng/ml)의 세포 성장촉진 효과가 용량의 증가에 따라 증가하는 경향을 보였다. genistein의 경우 1uM을 정점으로 세포성장촉진이 극에 달하다가 10uM 투여군에서 1uMxndurns에 비하여 약 40% 정도 세포성장이 감소하였으나 대조군에 비하여 여전히 세포의 성장을 촉진하였다. 이러한 결과는 bisphenol A와 genistein이 에스트로젠과 같이 에스트로젠 수용체와 결합하는 경로로 MCF-7세포의 성장을 촉진한 것으로 생각된다. 따라서 이러한 결과를 확인하기 위하여, 에스트로젠 의존적으로 유전자가 발현되는 유전자 중의 하나인 pS2 유전자의 발현을 살펴보았다. Genistein과 bispheno A가 pS2 유전자의 발현을 유도하는 것이 northern blot 분석에 의하여 관찰되었다. 또한, 21일령의 무흉선 마이스를 이용한 in vivo 연구에서, 무처치 대조군에 비하여 genistein과 bisphenol A를 투여한 마우스 군에서 유선의 발달과 둥-bud 의 형성이 유의하게 증가됨이 관찰되었다(p<0.05). 결론적으로, bispheno A와 genistein이 MCF-7 세포에서 에스트로젠 효과가 매우 낮은 농도에서도 나타날 수 있다는 것은 식품위생상 큰 의미를 갖고 있으며, 이러한 물질들이 실제 사람에서의 위해성 특히 성장기 유아의 위해성에 대하여 좀더 면밀히 연구되어야 할 것이다.광에 따른 입자의 분쇄보다는 마광시 구형의 입자가 소성변형으로 인해 flake형상으로 변하여 체질입도분석시 입도의 증가를 초래하였으며, 반면 철산화물은 마광에 따른 입자의 미세화가 발생함을 볼 수 있었다. 나. 철분구 외수 실험 1) 광양의 dust를 40분간 마광하여 심강(분급)실험을 행했을 때 Fe 99.17% 품위 철분말을 37.8% 회수할 수 있었다. 2) 재항의 C/F dust를 40분간 마광하여 심강(분급)실험을 행했을 때 Fe 98.38% 품위의 철분말을 44.42% 회수할 수 있었다. 3) 70 gauss 자석을 사용하여 자력선별을 행했을 때 +65-200 mesh 사이에서 Fe 품위 98% 이상의 철분말을 회수 할 수 있으나 회수율(14%)이 낮다.0^{\circ}C$의 경우 20시간(20時間)에 가장 색도(色度)가 높아 갈변반응속도(褐變反應速度)가 0.2로 나타났다. the esophageal mucous cells pf Bryzoichthys lysimus contained small amount of neutral mucin, while on the other hand a feww mucous cells contained small amount of neutral mucin and minimal amount of sialomucin. But the esophageal mucous cells of Takifugu pardalis contained considerable amount of neutral mucin only.분해가 더욱 촉진되었으며, 30℃에서 교반 처리를 행한 경우가 10℃에서 교반 처리를 행한 경우 보다 지방분해가 더욱 촉진되었다. 산양유 원유는 30℃에서 교반 처리 시간이 연장되어도 지방분해는 뚜렷한 증가를 나타내지 않았다.와 표준체중군 여자에게서 가장 높게 나타났다. 5. 남자의 53.9%, 여자의 83.2%가 체중 조절에 관심이 있다고

• 생명과학회지
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• 제26권3호
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• pp.275-281
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• 2016

인체치료용 당단백질은 바이오의약품에 있어서 중요한 요소이며 특히, 단백질 말단에 결합되어있는 시알산은 의약품의 체내 활성이나 안정성에 큰 영향을 미친다. 최근 돼지유즙은 바이오의약품을 생산하기 위한 생체반응기로써 주목받고 있다. β-1,4-N-acetylglucosaminyltransferase A (MGAT4A)는 재조합단백질이 가지는 시알산 함량을 늘리기 위한 필수 효소 중 하나이다. 그러나 돼지 MGAT4A는 아직도 그 서열이나 기능이 불분명하다. 따라서 이번 연구에서는 돼지 MGAT4A의 서열 동정 및 기능분석을 하였다. 돼지 MGAT4A는 535개의 아미노산을 코딩하는 1,638개의 염기서열로 이루어져 있으며, 일반적인 당전이효소들의 특징인 type II membrane topology의 특성을 가지고 있다. Real-time PCR분석법을 통하여, 지금까지 알려진 것과는 조금 다르게 MGAT4A 유전자가 간이나 유선에서 많은 발현을 보였으나 소장이나 위, 방광에서는 그 발현량이 적다는 것도 확인하였다. 또한, 효소의 기능 분석을 위하여 PK-15 세포주에서 MGAT4A효소의 과발현을 유도하였다. 과발현이 확인된 세포주로 렉틴을 이용한 면역형광염색법과 ELISA방법으로 MGAT4A효소의 과발현이 β-1,4-N-acetylglucosamine의 함량을 증가시켰음을 확인하였다. 또한 기질반응을 통해 bi-antennary structure에 대한 반응성도 확인하였으며, MGAT4A의 과발현이 유도된 세포에서 약 3배 이상의 높은 기질 반응성을 보였다. 이러한 연구는 생체반응기로써 돼지를 이용하는데 있어서 중요한 기반이 될 것이라고 생각한다.