• Reproductive and Developmental Biology
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• 제35권1호
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• pp.23-31
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• 2011

Two piglets and one juvenile pig were used to investigate closely what types of cells express green fluorescent protein (GFP) and if any, whether the GFP-tagged cells could be used for stem cell transplantation research as a middle-sized animal model in bone marrow cells of recloned GFP pigs. Bone marrow cells were recovered from the tibia, and further analyzed with various cell lineage markers to determine which cell lineage is concurrently expressing visible GFP in each individual animal. In the three animals, visible GFP were observed only in proportions of the plated cells immediately after collection, showing 41, 2 and 91% of bone marrow cells in clones #1, 2 and 3, respectively. The intensity of the visible GFP expression was variable even in an individual clone depending on cell sizes and types. The overall intensities of GFP expression were also different among the individual clones from very weak, weak to strong. Upon culture for 14 days in vitro (14DIV), some cell types showed intensive GFP expression throughout the cells; in particular, in cytoskeletons and the nucleus, on the other hand. Others are shown to be diffused GFP expression patterns only in the cytoplasm. Finally, characterization of stem cell lineage markers was carried out only in the clone #3 who showed intensive GFP expression. SSEA-1, SSEA-3, CD34, nestin and GFAP were expressed in proportions of the GFP expressing cells, but not all of them, suggesting that GFP expression occur in various cell lineages. These results indicate that targeted insertion of GFP gene should be pursued as in mouse approach to be useful for stem cell research. Furthermore, cell- or tissue-specific promoter should also be used if GFP pig is going to be meaningful for a model for stem cell transplantation.

• Environmental Analysis Health and Toxicology
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• 제19권2호
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• pp.141-151
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• 2004

We exposed juvenile puffer fish, Takifugu obscurus(30 days after hatching) to various aqueous pollutants including 4 kinds of inorganic metals (Ag, Cd, Cu and Hg), 2 organotin compound.; (tributyltin ［TBT］ and triphenyltin［TPhT］) and 5 polycyclic aromatic hydrocarbon (PAH) compounds (chrysene, fluoranthene, naphthalene, phenanthrene and pyrene) to estimate median lethal concentrations (LC50s) of each pollutant after the 96-hour acute exposure. Among the inorganic metals, Hg (52 $\mu\textrm{g}$/L; 96-h LC50) was most toxic to test animals and followed by Ag (164 $\mu\textrm{g}$/L), Cu (440 $\mu\textrm{g}$/L) and Cd (1180 $\mu\textrm{g}$/L). Aqueous TBT was more toxic between the two organotins; the 96-h LC50 for TBT (5.1 $\mu\textrm{g}$/L) was 3 times lower than that of TPhT (17.3 $\mu\textrm{g}$/L). The acute toxicity of PAH compounds was highest for chrysene (1.5 $\mu\textrm{g}$/L; 96-h LC50) and decreased in the order of pyrene (65 $\mu\textrm{g}$/L) > fluoranthene (158 $\mu\textrm{g}$/L) > phenanthrene (432 $\mu\textrm{g}$/L) > naphthalene (8690 $\mu\textrm{g}$/L). The toxicity of PAH compounds wat closely related to their physico-chemical characteristics such as $K_{ow}$ and water solubility, and well explained by simple QSAR relationship. The sensitivity of puffer fish to various inorganic and organic pollutants was generally comparable to various fish species widely used as standard test species in previous studies and further evaluation should be conducted to develop adequate testing procedure for T. obscurus when used in various toxicity tests.

• Asian-Australasian Journal of Animal Sciences
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• 제17권2호
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• pp.174-178
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• 2004

The objectives of this study were to investigate the ovarian responsiveness of juvenile calves to exogenous gonadotropin treatments and to establish the oocyte retrieval technique for prepubertal heifers. Three 78-day-old calves were treated with 4 doses (40, 30, 30 and 30 mg) of FSH (Folltropin V) at 12 h interval up to 229 day-old. Surgical oocyte retrieval was performed 24 h after the last injection of FSH. Calves with good ovarian responses to FSH treatment had an average ovarian size of $5{\times}3$ cm compared to $3{\times}2$ cm in the less-responsive animals. Large variations were observed in the number of total follicles ($51{\pm}45$), aspirated follicles ($39{\pm}36$), oocytes recovered ($23{\pm}25$) and usable oocytes recovered ($11{\pm}19$) during 78 to 229 day-old. Oocytes derived from prepubertal calves had significantly lower maturation rate than those from cows (34 vs. 100%, p<0.05). Mean diameters of calf oocytes ($144{\pm}1{\mu}m$) and ooplasm ($110{\pm}1 {\mu}m$) were significantly lower than those of cows ($149{\pm}1$ and $25{\pm}1{\mu}m$, respectively). The diameter of the ooplasm also increased significantly after in vitro maturation (IVM) ($108{\pm}1$ vs. $112{\pm}1{\mu}m$). However, further studies are required to optimize the IVP system for the oocytes derived from prepubertal heifers.

• 한국환경생물학회:학술대회논문집
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• 한국환경생물학회 2002년도 학술대회
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• pp.123-126
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• 2002

Pollutants that are persistent, bioaccurnulative, and toxic have been linked to numerous adverse effects in human and animals, PBTs include heavy metals, polychlorinated biphenyls (PCBs), dioxins, polycyclic aromatic compounds (PACs) in addition to pesticides. This study focuses on toxic effects of the PBTs except pesticides on insects. Eight PBTs were selected from subgroups: three heavy metals (Pb, Hg, and Cd), two PCB mixtures (Aroclor mixtures 1 and 2), 2,3,7,8-tetrachlorodibenzo-p-dioxin, two monophenols (4-octylphenol and 4-nonylphenol), and tetrabutyltin, Beet armyworm, Spodoptera exigua, was used as test target insect species. Three physiological markers (metamorphosis, immune reaction, and follicle patency) were assessed in each exposure to different doses of the PCBs. Heat-shock proteins as molecular markers were also analyzed in response to the PCBs. All tested PBTs were toxic to metamorphosis from larvae to pupae when they were applied with diet. Two PCB mixtures were the most toxic compounds in this assay by giving significant toxicity at 0.005 ppm, while others had from 10 to 1000 ppm. Dioxin (0.1 ppb), tetrabutyltin (0.1 ppb), Pb (10 ppb), and Hg (0,01 ppb) were potent to inhibit immune reactions analyzed by inducing phenoloxidase activity and blocked phospholipase $A_2$ enzyme, Tetrabutyltin and dioxin significantly induced follicle cell patency, but their effects were lower than that of endogenous juvenile hormone, Dioxin, Pb, Hg, and Cd could induce the expression of heat shock proteins that were detected by immunoblotting against human HSP70 monoclonal antibody. HSP78 and HSP80 were upregulated in response to the PBTs. This expression was detected from the fat body and epidermis at as fast as 4h after injection. All these results clearly suggest that PBTs give significant ecotoxicity to insects that are valuable organisms in our environment.

• Applied Microscopy
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• 제28권1호
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• pp.91-105
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• 1998

The present study is undertaken to investigate the hypertrophical changes of the corpus allatum (corpora allata, CA) after the ovariectomy in Blattella germanica. In particular, the ultrastructural aspects of the normal and ovariectomized conditions, and induced factors of the hypertrophic phenomenon are focused. Ultrastructure of the CA from an immediately emergent adult is similar to that of the last larval stage that has stopped secreting juvenile hormone. The CA is composed of undifferentiated cells, exhibiting an electron-lucent matrix, a few mitochondria and less smooth endoplasmic reticulum. The karyoplasm occupy most of the cytoplasm. Electron-dense materials are filled with the intercellular spaces and gap junctions are also found. Almost no ultrastructural changes have been noticed during seven days until the oviposition. However, considerable changes in structure have been detected soon after the oviposition. Mitochondria are increased dramatically in number and cristae, and changed to the filamentous form with a high electron density. In addition, Golgi complexes, microtubules, and polysomes are also increased. After an oviposition, the total volume of the CA are decreased again. The volume of the CA are increased continuously, hypertrophy, after the ovariectomy. Morphological aspects of the CA in an early stage after the removal are similar to the structure of the secondary egg maturation. Large and electron dense globules are observed in the ovariectomized CA cytoplasms and they are present in those cells for a long period of time. Yet such a hypertrophical phenomenon occur only in the specific cells. The hypertrophy are caused by hollowing the part of the CA cells and later filling such site with polysomes. In 42 days after the ovariectomy, the nuclear membranes disappear in the CA cells, thus, exhibiting the prokaryotic-like features. Some results of the current study will contribute to the establishment of the model that explain unusual changes accompanied by certain treatment in insects and/or further in animals.

• 한국동물위생학회지
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• 제45권4호
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• pp.249-252
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• 2022

Giardiasis is widespread all over the world, and it is a disease that causes both acute and chronic digestive symptoms. It is zoonotic disease that affects animals and humans. There are few studies on giardiasis in stray cats due to difficulties in catching and sampling. Therefore, this study evaluated the prevalence of giardiasis in stray cats in the Daejeon city because of increasing interest as zoonotic disease. The specimens were the feces of stray cats captured for the neutering project (TNR) in Daejeon; 30 fecal samples were collected from 2021 to 2022 in each of 5 districts in Daejeon. A total of 150 samples were collected. All samples were tested for giardiasis using the Giardia SNAP kit (SNAP^Ⓡ test, IDEXX Laboratories. Inc., Westbrook, ME). The overall prevalence rate was 46 out of 150 cats (30.7%). By age, 25 out of 71 juvenile cats (35.2%) were positive, and 21 out of 79 adult cats (26.6%) were positive. A total of 19 out of 69 cats (27.5%) with diarrhea were positive, and 27 out of 81 asymptomatic cats (33.3%) were positive. For gender, 38 out of 99 females (38.4%) were positive, and 8 out of 51 males (15.7%) were positive. The positive rate of giardiasis in stray cats was over 30%, which is high compared to other research results. It is necessary to increase the public's awareness of the value of deworming stray cats and the sanitation of people who have come into contact with them.

• 한국발생생물학회지:발생과생식
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• 제15권4호
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• pp.365-371
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• 2011

수온은 어류를 포함한 수중동물의 다양한 생물학적 사건에 영향을 미친다. 어류의 번식 및 발생도 수온의 변화와 밀접한 관련이 있다. 어류를 높은 수온에 노출시키면 HSP70 유전자 발현을 유도한다. 따라서, HSP70 유전자 발현을 효과적으로 차단할 수 있는 저해제의 개발은 온도에 민감한 생물학적 사건과 관련된 HSP70의 역할을 연구하는데 크게 기여할 수 있다. 본 연구에서는 고온($36^{\circ}C$)에 노출된 틸라피아 자어와 치어(전장 10~13 cm)에서 천연 flavonoid의 일종인 quercetin, 3,3',4',5,7-pentahydroxyflavon의 "고온 유도 HSP70 유전자 발현" 억제효과를 조사하였다. 고온 노출 전에 50 ${\mu}M$ 이상의 quercetin 용액에 6시간 동안 사전 침지한 실험어에서는 HSP70 유전자 발현이 유의하게 감소하였다(P<0.05). 특히, 100 ${\mu}M$로 침지 처리한 개체에서는 고온 처리를 하지 않은 실험어에서 만큼 낮은 HSP70 유전자 발현을 나타내었다. 한편, 0.1 $m{\ell}$씩의 0.5 mM, 5 mM 또는 20 mM의 quercetin 용액을 치어에 주사한 결과, 고수온 유도 HSP70 유전자 발현은 0.5 mM의 quercetin을 주사한 치어의 생식소와 뇌에서 모두 유의하게 감소되었다(P<0.05). 본 연구의 결과는 quercetin이 어류에서 고온에 의해 유도된 HSP70 유전자 발현을 효과적으로 차단할 수 있으며, 틸라피아 자어 침지의 경우 유효 농도 100 ${\mu}M$ 그리고 치어 주사의 경우 유효 농도 0.5 mM로 원하는 효과를 얻을 수 있음을 제시한다.

• 대한수의학회지
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• 제14권1호
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• pp.115-133
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• 1974

Although considerable research has been done on the changes associated with age in the blood picture of domestic and laboratory animals, little work has been made of the changes occurring at different age in the blood picture of goats. And a comprehensive survey of the bood picture of Korean native goats has not been made. The object of the present investigation was to suggest standards for the blood picture of Korean native goats at frequent intervals from birth to maturity. The goats were kept under average farming conditions in this country. Observations were made at the following ages: at birth; 2,4 and 7 days; 2, 3, 4, 5, 6, 7, 8 and 9 weeks; 2.5, 3,6 and 12 months. Blood samples were drawn from the jugular vein. It was taken between 8 and 9 a.m. except those for the at-birth period. Erythrocyte and leukocyte enumerations and, determinations of hemoglobin in blood and hematocrit value were made in the usual manner. Reticulocytes were enumerated per 1,000 erythrocytes in blood smears stained with briIliant cresyl blue and counterstained with Wright's stain. Erythrocytes counts declined from $8.7{\times}10^8/mm^3$ at birth to a low of 7.0 at 4 days of age. These values increased to 11.5 at 5 weeks and reached a maximum of 14.0 at 3 months of age; it then fell to 11.5 at 12 months of age. Concentrations of hemoglobin in blood and hematocrit values were not related to the changes of erythrocyte counts. The values at birth were higher than at any other period during the first year of life. These fell from highs of 12.3 g/100 ml and 38.0 ml/100 ml to lows of 9.2 and 29 at 4 weeks for concentration of hemoglobin in blood and hematocrit value, respectively. There was a common pattern for the hematocrit value and hemoglobin in blood which showed three phases-a fall during the first month, a rise to the third month, and a fall to the mature level at 12 months of age. Mean corpuscular volume and mean corpuscular hemoglobin showed a common pattern. The values were $44.2{\mu}m^3$ and 14.2 pg at birth and fell, at first slowly and then rapidly, to reach adult levels of 24.1 and 7.9 at 6 weeks of age for mean corpuscular volume and mean corpuscular hemoglobin, respectively. Mean corpuscular hemoglobin concentration was little affected by age. Reticulocyte was observed from birth to 4 weeks of age. Percentage of reticulocyte decreased from 0.85% at birth to 0.06% at 4 weeks of age. Total leucocyte counts increased from $5.64{\times}10^3/mm^3$ at birth to a maximum of 13.4 at 3 months; it then fell to 11.5 at 12 months of age. In differential counts myelocyte, juvenile and band form decreased with advancing age. No myelocyte and juvenile were seen after the age of 7 and 9 weeks, respectively, and band forms were rare after the age of 3 months. Percentage of mature neutrophil showed a quick decline from 52.5% at birth to reach a minimum level (34.5%) at 3 months of age; it then rose to 38% at 12 months of age. Percentage of lymphocyte increased from 39.2% at birth to maximum of 59% at 3 month of age; it then fell to 54.9% at 12 months of age. Percentage of monocyte was not affected with advance of age. Percentage of eosinophil and basophil were increased with advance of age to reach a maximum at 2 to 3 month of age. It then fell to adult level at 12 month of age.

• 한국환경생태학회지
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• 제28권5호
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• pp.523-530
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• 2014

2011년 6월 28일에 파주시 조리읍 장곡리 인근 도로에서 두부쪽이 손상되고, 해당연도에 태어난 맹금류 어린새를 발견하였고, 이를 DNA 바코드 기법으로 동정하였다. Mitochondrial DNA(mtDNA) cytochrome c oxidase I(COI) 유전자의 695bp 절편을 중합효소연쇄반응(polymerase chain reaction, PCR)으로 증폭하고 염기서열을 결정하였다. 결정된 염기서열을 BOLD systems과 NCBI의 BLAST에서 유사도 분석을 수행한 결과 총 5개체의 왕새매가 검색되었고, 염기서열의 동일성은 100%로 조사되었다. 또한, DNA 분자성판별 결과는 해당 개체가 암컷임을 나타내었다. 이러한 결과는 경기도 파주에서 1968년 이후 43년만에 왕새매의 번식이 확인된 중요한 정보로, 향후 광역야생동물구조센터는 야생동물의 사체 수거 시 인근 기탁등록보존기관과 연계하여 DNA시료를 확보하고 보다 정확한 종동정과 성판별 정보를 기록하는 등의 체계적인 관리시스템이 필요할 것으로 사료된다. 또한 이렇게 확보한 왕새매의 DNA 시료와 DNA 바코드 COI 유전자 서열은 유사종 연구의 참조표본(reference standard)로 이용될 수 있을 것이다.

• Applied Microscopy
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• 제36권2호
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• pp.93-99
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• 2006

Gemifloxacin은 대부분의 G(-) 세균들과 G(+) 세균들에 대해 강한 활성을 나타내며 미성숙동물에서 비교적 낮은 연골독성 가능성을 가진 합성 fluoroquinolone계 항생제이다. 본 연구는 미성숙 SD계 랫드의 아킬레스건에 gemifloxacin이 미치는 영향을 평가하기 위해 gemifloxacin을 생후 4주 후부터 0(vehicle), 600mg/kg body weight 용량으로 5일 동안 매일 한번씩 경구투여를 하여 ofloxacin과 비교하였다. 아킬레스건의 형태학적 변화를 투과전자현미경을 이용하여 관찰한 결과 부형제만 처치한 대조군과 비교하였을 때, gemifloxacin과 ofloxacin을 처치한 랫드로부터 채취한 샘플에서 초미세구조의 변화가 관찰되었다. quinolone을 처치한 군의 tenocyte에서는 다양한 퇴행성 변화가 관찰되었고, 세포외기질에서 분리된 세포들이 존재하였는데 이는 ofloxacin을 처치한 그룹에서 특히 높았고 gemifloxacin을 처치한 그룹에서는 매우 낮았다. 따라서, gemifloxacin이 아킬레스건과 같은 결합조직 구조물들의 퇴행성 변화를 보다 적게 야기한다는 것을 보여준다.