• Asian-Australasian Journal of Animal Sciences
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• 제31권9호
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• pp.1420-1430
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• 2018

Objective: Epigallocatechin-3-gallate (EGCG) is a major ingredient of catechin polyphenols and is considered one of the most promising bioactive compounds in green tea because of its strong antioxidant properties. However, the protective role of EGCG in bovine oocyte in vitro maturation (IVM) has not been investigated. Therefore, we aimed to study the effects of EGCG on IVM of bovine oocytes. Methods: Bovine oocytes were treated with different concentrations of EGCG (0, 25, 50, 100, and $200{\mu}M$), and the nuclear and cytoplasmic maturation, cumulus cell expansion, intracellular reactive oxygen species (ROS) levels, total antioxidant capacity, the early apoptosis and the developmental competence of in vitro fertilized embryos were measured. The mRNA abundances of antioxidant genes (nuclear factor erythriod-2 related factor 2 [NRF2], superoxide dismutase 1 [SOD1], catalase [CAT], and glutathione peroxidase 4 [GPX4]) in matured bovine oocytes were also quantified. Results: Nuclear maturation which is characterized by first polar body extrusion, and cytoplasmic maturation characterized by peripheral and cortical distribution of cortical granules and homogeneous mitochondrial distribution were significantly improved in the $50{\mu}M$ EGCG-treated group compared with the control group. Adding $50{\mu}M$ EGCG to the maturation medium significantly increased the cumulus cell expansion index and upregulated the mRNA levels of cumulus cell expansion-related genes (hyaluronan synthase 2, tumor necrosis factor alpha induced protein 6, pentraxin 3, and prostaglandin 2). Both the intracellular ROS level and the early apoptotic rate of matured oocytes were significantly decreased in the $50{\mu}M$ EGCG group, and the total antioxidant ability was markedly enhanced. Additionally, both the cleavage and blastocyst rates were significantly higher in the $50{\mu}M$ EGCG-treated oocytes after in vitro fertilization than in the control oocytes. The mRNA abundance of NRF2, SOD1, CAT, and GPX4 were significantly increased in the $50{\mu}M$ EGCG-treated oocytes. Conclusion: In conclusion, $50{\mu}M$ EGCG can improve the bovine oocyte maturation, and the protective role of EGCG may be correlated with its antioxidative property.

• Applied Microscopy
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• 제29권4호
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• pp.417-426
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• 1999

농어의 피부는 크게 상피층과 진피층으로 구분된다. 상피층은 지지세포와 단세포선들로 구성된다. 지지세포는 표면세포, 중간세포 및 기저세포로 구분된다. 선세포는 점액분비세포와 곤봉상세포로 구분되는데, 이들의 분포밀도는 곤봉상세포가 더 높다. 표면세포는 편평형 또는 입방형이며 세포질에는 조면소포체가 잘 발달되어 있으며, microridge가 뚜렷하다. 표면세포들은 membrane interdigitation구조와 부착반에 의해서 다른 세포들과 연결되어 있다. 중간세포는 난형이며, 전자밀도는 다른 지지세포 보다 높다. 기저세포는 입방형이며, 세포소기관의 가운데 미토콘드리아의 발달이 현저하며, membrane interdigitation 구조들이 발달되어 있다. 점액분비세포는 막으로 싸인 다수의 분비과립들을 함유한다. 곤봉상세포의 세포질은 수질부와 피질부로 구분되는데, 수질부에서는 핵, 세포소기관, 중심공포가 관찰되며, 피질부에서는 발달된 장미세섬유들이 관찰된다. 곤봉상세포들은 잘 발달된 membrane interdigitation구조와 부착반에 의해서 다른 세포들과 연계되어 있다.

• 한국식품과학회지
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• 제27권6호
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• pp.915-920
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• 1995

This study was performed to investigate the effect of aluminum compound on the aluminum contents and histological change in brain tissue of rats. Seventy five male Sprague-Dawley strains were divided into five groups consisting of the control, 250 ppm $AlCl_3$ group, 500 ppm $AlCl_3$ group, 250 ppm $Al_2(SO_4)_3$ group, 500 ppm $Al_2(SO_4)_3$ group and kept on the diet for 2 weeks. The weight gain was increased by administration of $AlCl_3$ but decreased by administration of $Al_2(SO_4)_3$ as compared to control group. The aluminum contents in brain tissue of each group; 250 ppm $AlCl_3$ group, 500 ppm $AlCl_3$ group, 250 ppm $Al_2(SO_4)_3$ group and 500 ppm $Al_2(SO_4)_3$ group were 64.63, 102.21, 132.64 and 180.41 ppm, respectively. Aluminum accumulation in brain tissue was higher with administration of $Al_2(SO_4)_3$ than with administration of $AlCl_3$. In $AlCl_3$ administration group, multiple small intracytoplasmic granules and microvacuole were seen in large pyramidal cells of cortex and granulovacuolar degeneration. In $Al_2(SO_4)_3$ administration group revealed pollagis pallor, cellular pyknosis, microcavitation resulted from edema in deeper cortical layers were observed. Blue-pigmentation which represents the accumulation of aluminum was noted In granulovacuolar degeneration site in $Al_2(SO_4)_3$ administration group.

• Molecules and Cells
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• 제40권8호
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• pp.558-566
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• 2017

Regular monitoring on experimental animal management found the fluctuation of ART outcome, which showed a necessity to explore whether superovulation treatment is responsible for such unexpected outcome. This study was subsequently conducted to examine whether superovulation treatment can preserve ultrastructural integrity and developmental competence of oocytes following oocyte activation and embryo culture. A randomized study using mouse model was designed and in vitro development (experiment 1), ultrastructural morphology (experiment 2) and functional integrity of the oocytes (experiment 3) retrieved after PMSG/hCG injection (superovulation group) or not (natural ovulation; control group) were evaluated. In experiment 1, more oocytes were retrieved following superovulation than following natural ovulation, but natural ovulation yielded higher (p < 0.0563) maturation rate than superovulation. The capacity of mature oocytes to form pronucleus and to develop into blastocysts in vitro was similar. In experiment 2, a notable (p < 0.0186) increase in mitochondrial deformity, characterized by the formation of vacuolated mitochondria, was detected in the superovulation group. Multivesicular body formation was also increased, whereas early endosome formation was significantly decreased. No obvious changes in other microorganelles, however, were detected, which included the formation and distribution of mitochondria, cortical granules, microvilli, and smooth and rough endoplasmic reticulum. In experiment 3, significant decreases in mitochondrial activity, ATP production and dextran uptake were detected in the superovulation group. In conclusion, superovulation treatment may change both maturational status and functional and ultrastuctural integrity of oocytes. Superovulation effect on preimplantation development can be discussed.

• 한국한의학연구원논문집
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• 제8권1호
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• pp.93-104
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• 2002

The following is a list of morphologic and genetic characteristics of Pinellia tuber. 1. The original plant of Pinellia tuber is Pinellia ternata$(T_{HUNB})$$B_{REIT}$. With regards to its external morphology, it is smaller than other Araceae species and its spadix is longer than its leaves, which trifurcate. 2. As regards its internal morphology, its mucous cell is elliptical and the vessel is helical or annular-shaped. Granules exist in abundance and in various shapes. 3. Distribution and size of laticifers are the key criteria on which to differentiate between domestic and imported Pinellia tuber. Laticifers are mainly distributed in the epidermis in domestic Pinellia tuber and in the cortical parenchyma in imported Pinellia tuber. The size of laticifers is somewhere between 1,3 and $8{\mu}m$ in diameter in imported Pinellia tuber bigger than its domestic counterpart. 4. RAPD markers display a great similarity in bands between domestic and Chinese Pinellia tuber. However, RAPD primers 352, 358, 365, 368 and 374 are distinctive markers for domestic Pinellia tuber. In the meantime, North Korean Pinellia tuber, morphologically similar to domestic Pinellia tuber, is genertically distinctive from its domestic counterpart in primers 354, 358, 365, 368, 374 and 379, a finding that supports the postulation that North Korean Pinellia tuber is tuber of another Araceae species.

• 농업과학연구
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• 제48권3호
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• pp.607-615
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• 2021

Cytokines are protein mediators that possess the ability to assist cell-to-cell communication in immune system-related activities. In general, pathogen endotoxins activate the release of inflammatory mediators, and with time, there is an increase in the cytokine levels in the body. Interleukin (IL)-6 mediates the acute-phase inflammatory response, and elevated IL-6 levels have been reported in peritoneal fluids of women with pelvic inflammation and endometriosis, thereby associating it with oocyte quality and infertility. To overcome subfertility or infertility in humans and animals, the present study was done to examine the effect of recombinant IL-6 on porcine oocytes matured in vitro and subsequently to determine the fertilization rate and embryo development. Porcine oocytes were incubated with varying concentrations of IL-6 (0 - 2 ㎍·mL^-1) for 44 h followed by in vitro fertilization and culturing of the oocytes. The oocytes or embryos were fixed with 3.7% paraformaldehyde (PFA) and stained with fluorescence dyes, and the meiotic spindle, chromosome organization, fertilization status and embryo development were subsequently assessed under a fluorescence microscope. We observed induction of an abnormal meiotic spindle alignment in the oocytes incubated with IL-6 compared to the control oocytes incubated without IL-6. Moreover, significantly decreased fertilization rates and embryo development were observed for oocytes incubated with IL-6 (p < 0.05). Thus, an increased IL-6 level during oocyte maturation could be associated with fertilization failure due to an aberrant chromosomal alignment and a disruption of the cortical granules. Taken together, our results indicate that successful assisted reproduction can be achieved by controlling the levels of inflammatory cytokines.

• 한국임상수의학회지
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• 제26권6호
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• pp.586-590
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• 2009

철선충은 주로 사마귀와 같은 곤충에 기생하는 선충류로 사람이나 동물은 우연히 감염될 수 있는 것으로 알려져 왔다. 철선충의 생활사는 크게 곤충에 기생하는 기생생활기, 그리고 발육이 완료된 후에 숙주를 죽이고 외계로 나와 자유생활을 하는 자유생활기로 구분할 수 있다. 본 연구는 개의 구토물에서 오디흑연가시 수컷을 발견하여 이의 횡단면을 주사 및 투과 전자현미경으로 관찰한 것이다. 전자현미경하에서 표피는 4층(epicuticle, cuticle, epidermis, muscle)으로 이루어졌고, 내강에는 장, 배신경절이 발견되었다. 그러나 고환은 발견되지 않았다. 표피의 최상층에서 여러 형태의 areoles이 관찰되었으며, cuticle은 17열의 섬유로 이루어져 있었다. Epidermis는 1열로 구성되어 있었으며, muscle층은 여러 개의 근세포가 종으로 일정하게 배열되어 있었다. 그리고 내강에 배신경절이 충체의 정중앙 배측에서만 존재하는 것으로 확인되었다. 이러한 결과로 본 철선충은 기생생활기를 끝내지 못한 미성숙충으로 판단된다.

• Asian-Australasian Journal of Animal Sciences
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• 제32권8호
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• pp.1112-1121
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• 2019

Objective: Gram-negative bacteria lipopolysaccharide (LPS) has been reported to be associated with uterine impairment, embryonic resorption, ovarian dysfunction, and follicle retardation. Here, we aimed to investigate the toxic effects of LPS on the maturation ability and parthenogenetic developmental competence of bovine oocytes. Methods: First, we developed an in vitro model to study the response of bovine cumulusoocyte complexes (COCs) to LPS stress. After incubating germinal vesicle COCs in $10{\mu}g/mL$ of LPS, we analyzed the following three aspects: the expression levels of the LPS receptor toll-like receptor 4 (TLR4) in COCs, activities of intracellular signaling protein p38 mitogen-activated protein kinase (p38 MAPK) and nuclear factor-kappa B (NF-${\kappa}B$); and the concentrations of interleukin (IL)-$1{\beta}$, tumor necrosis factor (TNF)-${\alpha}$, and IL-6. Furthermore, we determined the effects of LPS on the maturation ability and parthenogenetic developmental competence of bovine oocytes. Results: The results revealed that LPS treatment significantly elevated TLR4 mRNA and protein expression levels in COCs. Exposure of COCs to LPS also resulted in a marked increase in activity of the intracellular signaling protein p-p38 MAPK and NF-${\kappa}B$. Furthermore, oocytes cultured in maturation medium containing LPS had significantly higher concentrations of the proinflammatory cytokines IL-$1{\beta}$, TNF-${\alpha}$, and IL-6. LPS exposure significantly decreased the first polar body extrusion rate. The cytoplasmic maturation, characterized by polar body extrusion and distribution of peripheral cortical granules, was significantly impaired in LPS-treated oocytes. Moreover, LPS exposure significantly increased intracellular reactive oxygen species levels and the relative mRNA abundance of the antioxidants thioredoxin (Trx), Trx2, and peroxiredoxin 1 in oocytes. Moreover, the early apoptotic rate and the release of cytochrome C were significantly increased in response to LPS. The cleavage, morula, and blastocyst formation rates were significantly lower in parthenogenetically activated oocytes exposed to LPS, while the incidence of apoptotic nuclei in blastocysts was significantly increased. Conclusion: Together, these results provide an underlying mechanism by which LPS impairs maturation potential in bovine oocytes.

• Applied Microscopy
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• 제7권1호
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• pp.21-43
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• 1977

This experiment was undertaken in order to localize the labeled dbcAMP (dibutyryl cyclic AMP) in oocytes whose development has been suppressed by cold dbcAMP for 6 or 19 hours in vitro. Mouse oocytes were obtained from the ovaries of 3-4 week old A strain female mice, by puncturing the Graafian follicles in the modified Krebs-Ringer bicarbonate salt solution under the dissecting microscope. Those oocytes which have intact germinal vesicle were cultured in the basic culture medium supplemented with 0.4% bovine serum albumin (BSA). Cultivation of the oocytes was carried out in a microtube developed by Cho (1974). The cultures were then incubated in a humidified 5% $CO_2$ incubator maintained at $37^{\circ}C$ for 6 or 19 hours (Donahue, 1968). DbcAMP was added to culture medium for a final concentration of 100ug/ml, and $^3H-dbc$ AMP (specific activity 13 Ci/mM) for a final concentration of $40{\mu}Ci/ml$ was also added to the medium. For electron microscopic autoradiography, those oocytes recovered from the culture were washed with phosphate buffer (pH 7.4), and immediately prefixed in a 2.5% glutaraldehyde overnight and postfixed for 2 hours at $4 ^{\circ}C$ in 1% osmium tetroxide in phosphate buffer with pH 7.4 (Palade, 1952). After fixation, the materials were dehydrated in graded alcohol series and embedded in Epon 812 mixture based on the standard procedures (Luft, 1961). The thin sections $600-700{\AA}$ thick were mounted on the grids of 200 meshes. The grids containing sections were coated with a nuclear emulsion Kodak NTB-3 and stored in a cold dark box (at $4^{\circ}C$) for 3 weeks. After exposure, the samples were developed with Kodak D-19 and stained with uranyl acetate and lead citrate. Routine observation was made with Hitachi HU-11E electron microsocope. The results of the observation were as followings: 1. It was found that the labeled dbcAMP penetrated the egg plasma membrane and dispersed at random in the cytoplasm. 2. It was also observed that most of the labeled dbcAMP was attached to microfibrillar lattices portion of the oocyte cytoplasm. There fore, it is presumed that the receptor of the dbcAMP is localized in the microfibrillar lattices of the oocyte. 3. It also seems that some other cell organells such as mitochondria, Golgi complex, cortical granules are not directly related to the action of the dbcAMP. 4. The labeled dbcAMP was neither observed in the membrane nor in the nucleus. Therefore, it seems that there is no relationship between the concentration of dbcAMP and the nuclear membranous permeability. 5. There was no difference in number of dbcAMP particles when oocytes were cultured for 6 hours and 19 hours. 6. However, it was observed that, in same of the oocytes suppressed in germinal vesicle by dbcAMP for 19 hours, cell organells were moved and concentrated to a small portion of the cytoplasm, and that the morphology of the organells greatly changed to an abnormal. form. Therefore, it is supposed that those oocytes were in the process of degeneration. From the above results, it is expected that dbcAMP penetrated the egg membrane and was bound to the receptor which seems to be located in the microfibrillar lattiees portion, and that this dbcAMP-receptor complex inhibited some enzyme system of the oocytes which are essential for the germinal vesicle breakdown.