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Effects of Different Types of Dietary Fat on Muscle Atrophy According to Muscle Fiber Types and PPAR${\delta}$ Expression in Hindlimb-Immobilized Rats (지방의 종류가 다른 식이의 섭취가 하지고정 흰 쥐의 근 섬유별 근 위축과 PPAR${\delta}$ 활성에 미치는 영향)

  • Lee, Ho-Uk;Park, Mi-Na;Lee, Yeon-Sook
    • Journal of Nutrition and Health
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    • v.44 no.5
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    • pp.355-365
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    • 2011
  • This study investigated how dietary fat affects muscle atrophy and lipid metabolism in various muscles during hindlimb immobilization in rats. Twenty-four male Sprague?Dawley rats had their left hindlimb immobilized and were divided into four groups by dietary fat content and composition. The contralateral hindlimb (control) was compared with the immobilized limb in all dietary groups. Rats (n = 6/group) were fed a 4% corn oil diet (CO), 2.6% corn oil + 1.4% fish oil diet (FO), 30% corn oil diet (HCO), or a 30% beef tallow diet (HBT)after their hind limbs were immobilized for 10 days. Data were collected for the gastrocnemius, plantaris and soleus muscles. Muscle atrophy was induced significantly after 10 days of hindlimb immobilization, resulting in significantly decreased muscle mass and total muscle protein content. The protein levels of peroxisome proliferator activated receptor ${\delta}$ (PPAR${\delta}$) in the plantaris, gastrocnemius, and soleus increased following hindlimb immobilization irrespective of dietary fat intake. Interestingly, the PPAR${\delta}$ mRNA level in the plantaris decreased significantly in all groups and that in the FO group was lower than that in the other groups. The soleus PPAR${\delta}$ mRNA level decreased significantly following hindlimb immobilization in the FO group only. Muscle carnitine palmitoyl transferase 1 (mCPT1) mRNA level was not affected by hindlimb immobilization. However, the mCPT1 mRNA level in the FO group was significantly lower in the plantaris but higher in the soleus than that in the other groups. The pyruvate dehydrogenase kinase 4 (PDK4) mRNA level in the plantaris decreased significantly, whereas that in the soleus increased significantly following hindlimb immobilization. The plantaris, but not soleus, PDK4 mRNA level was significantly higher in the FO group than that in the CO group. The increased PPAR${\delta}$ protein level following hindlimb immobilization may have suppressed triglyceride accumulation in muscles and different types of dietary fat may have differentially affected muscle atrophy according to muscle type. Our results suggest that ${\omega}$-3 polyunsaturated fatty acids may suppress muscle atrophy and lipid accumulation by positively affecting the expression level and activity of PPAR${\delta}$ and PPAR${\delta}$-related enzymes, which are supposed to play an important role in muscle lipid metabolism.

Research and Development Trends on Omega-3 Fatty Acid Fortified Foodstuffs (오메가 3계 지방산 강화 식품류의 연구개발 동향)

  • 이희애;유익종;이복희
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.26 no.1
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    • pp.161-174
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    • 1997
  • Omega-3 fatty acids have been major research interests in medical and nutritional science relating to life sciences since after the epidemiologic data on Green3and Eskimos reported by several researchers clearly showed fewer per capita deaths from heart diseases and a lower incidence of adult diseases. Linolenic acid(LNA) is an essential fatty acid for human beings as well as linoleic acid(LA) due to the fact that vertebrates lack an enzyme required to incorporate a double bond beyond carbon 9 in the chain. In addition the ratio of omega-6 and 3 fatty acids seems to be important in terms of alleviation of heart diseases since LA and LNA competes for the metabolic pathways of eicosanoids synthesis. High consumption of omega-3 fatty acids in seafoods may control heart diseases by reducing blood cholesterol, triglyceride, VLDL, LDL and increasing HDL and by inhibiting plaque development through the formation of antiaggregatory substances like PGI$_2$, PGI$_3$ and TXA$_3$ metabolized from LNA. Omega 3 fatty acids also play an important role in neuronal developments and visual functioning, in turn influence learning behaviors. Current dietary sources of omega-3 fatty acids are limited mostly to seafoods, leafy vegetables, marine and some seed oils and the most appropriate way to provide omega-3 fatty acids is as a part of the normal dietary regimen. The efforts to enhance the intake of omega-3 fatty acids due to several beneficial effects have been made nowadays by way of food processing technology. Two different ways can be applied: one is add Purified and concentrated omega-3 fatty acids into foods and the other is to produce foods with high amounts of omega-3 fatty acids by raising animals with specially formulated feed best for the transfer of omega-3 fatty acids. Recently, items of manufactured and marketed omega-3 fatty acids fortified foodstuffs are pork, milk, cheese, egg, formula milk and ham. In domestic food market, many of them are distributed already, but problem is that nutritional informations on the amounts of omega-3 fatty acids are not presented on the labeling, which might cause distrust of consumers on those products, result in lower sales volumes. It would be very much wise if we consume natural products, result in lower sales volumes. It would be very much wise if we consume natural products high in omega-3 fatty acids to Promote health related to many types of adult diseases rather than processed foods fortified with omega-3 fatty acids.

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A Study on Garden Design Principles in "Sakuteiki(作庭記)" - Focused on the "Fungsu Theory"(風水論) - (「사쿠테이키(作庭記)」의 작정원리 연구 - 풍수론(風水論)을 중심으로 -)

  • Kim, Seung-Yoon
    • Journal of the Korean Institute of Landscape Architecture
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    • v.41 no.6
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    • pp.1-19
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    • 2013
  • This study tries to review 'Sakuteiki(作庭記)', the Book of Garden Making, compiled at the end of the 11th Century during the Heian Period of Japan, from the East-Asian perspective. 'Sakuteiki' is a Garden Theory Book, the oldest in the world as well as in Asia, and it contains the traditional knowledge of Japanese ancient garden culture, which originated from the continent(Korea and China). Traditional knowledge related to East-Asian garden culture reviewed in this paper is "Fungsu Theory"(風水, Asian traditional ecology: Fengshui in Chinese; Fusui in Japanese), stemmed from the culture to seek sound and blessed places to live in. Viewed from modern landscape architecture, the Fungsu Theory corresponds to ecology(science). The Fungsu Theory was established around the Han Dynasty of China together with the Yinyangwuxing(陰陽五行) Theory and widely used for making human residences including gardens. It was transmitted to Japan via Korea as well as through direct transaction between Japan and China. This study reinterprets garden design principles represented in Sakuteiki, which were selected in 5 key words according to the Fungsu Theory. The 5 key words for the Fungsu Theory are "the place in harmony of four guardian gods(四神相應地)", "planting trees in the four cardinal directions", "flow of Chi(氣)", "curved line and asymmetry", and "mountain is the king, water is the people". Garden design principles of "the place in harmony of four guardian gods(四神相應地)" and "planting trees in the four cardinal directions" are corresponding to "Myeongdang-ron(明堂論, Theory of propitious site)". The place in harmony of four guardian gods mentioned in Sakuteiki is a landform surrounded by the flow of water to the east, the great path to the west, the pond to the south, and the hill to the north. And the Theory originated from Zhaijing(宅經, Classic of dwelling Sites) of China. According to this principle, the city was planned and as a miniature model, the residence of the aristocrat during the Heian period was made. At the residence the location of the garden surrounded by the four gods(the flow of water, the great path, the pond, and the hill) is the Myeongdang(明堂, the propitious site: Mingtang in Chinese; Meido in Japanese). Sakuteiki explains how to substitute for the four gods by planting trees in the four cardinal directions when they were not given by nature. This way of planting originated from Zhaijing(宅經) and also goes back to Qiminyaoshu (齊民要術), compiled in the 6th Century of China. In this way of planting, the number of trees suggested in Sakuteiki is related to Hetu(河圖) and Luoshu(洛書), which are iconography of Yi(易), the philosophy of change, in ancient China. Such way of planting corresponds to that of Yongdoseo(龍圖墅, the villa based on the principle of Hetu) presented in Sanrimgyeongje (山林經濟), an encyclopedia on agriculture and living in the 17th Century of Korea. And garden design principles of "the flow of Chi(氣)", "curved line and asymmetry" is connected to "Saenggi Theory(生氣論, Theory of vitality)". Sakuteiki explains the right flow of Chi(氣) through the proper flow and the reverse flow of the garden stream and also suggests the curved line of the garden stream, asymmetric arrangement of bridges and stones in the garden, and indented shape of pond edges, which are ways of accumulating Chi(氣) and therefore lead to "Saenggi Theory" of the Fungsu Theory. The last design principle, "mountain is the king, water is the people", is related to "Hyeongguk Theory(形局論, Theory of form)" of the Fungsu Theory. Sakuteiki explains the meaning of garden through a metaphor, which views mountain as king, water as the people, and stones as king's retainers. It compares the situation in which the king governs the people with the help of his retainers to the ecological phenomena in which mountain(earth) controls water with the help of stones. This principle befits "Hyeongguk Theory(形局論, Theory of form)" of the Fungsu Theory which explains landform on the analogy of social systems, people, animals and things. As above, major garden design principles represented in Sakuteiki can be interpreted in the context of the Fungsu Theory, the traditional knowledge system in East Asia. Therefore, we can find the significance of Sakuteiki in that the wisdom of ancient garden culture in East-Asia was integrated in it, although it described the knowhow of a specific garden style in a specific period of Japan.

The Absorption and Purification of Air Pollutants and Heavy Metals by Selected Trees in Kwangju (광주지역(光州地域)에서 주요(主要) 수목(樹木)의 대기오염물질(大氣汚染物質)과 중금속(重金屬) 흡수(吸收) 정화기능(淨化機能)에 관(關)한 연구(硏究))

  • Cho, Hi Doo
    • Journal of Korean Society of Forest Science
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    • v.88 no.4
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    • pp.510-522
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    • 1999
  • The air pollutants ; $SO_2$, $SO{_4}^{-2}$, $NO{_3}^-$, $Cl^-$ are absorbed into soils through falling with dusts and rain from the atmosphere. The sources of heavy metal contaminants in the environments are agricultural and horticultural materials, sewage sludges, fossil fuel combustion, metallurgical industries, electronics and waste disposal etc.. The soils and hydrosphere can be polluted on the way of the circulation of these heavy metals. Studied pollutant anions are $SO{_4}^{-2}$, $NO{_3}^-$ and $Cl^-$ and heavy metals are Se, Mo, Zn, Cd, Pb, Mn, Cr, Co, V, As, Cu and Ni which are the elements to be concerned with the essentials for plants, with animal and human health. This study is with the aim of selecting the species of roadside trees and green space trees which have excellent absorption of air pollutants and heavy metals from the atmosphere and the soils in the urban area. Two areas are designated to carry out this study : urban area ; Kwangju city and rural area ; the yard of Forest Environment Institute of Chollanam-do, at Sanje-ri, Sampo-myum, Naju city, Chollanam-do (23km away from Kwangju). This study is carried out to understand the movement of anions and heavy metals from the soils to the trees in both areas, the absorption of anions and heavy metals from atmosphere into leaves and the amounts of anions and heavy metals in leaves and fine roots(< 1mm dia.) of roadside trees and green space trees in Kwangju and trees in the yard of Forest Environment Institute of Chollanam-do. The tree species selected for this study in both areas are Ginkgo biloba, Quercus acutissima, Cedrus deodara, Platanus occidentalis, Robinia pseudoacacia, Alnus japonica. Metasequoia glyptostroboides. Zekova serrata. Prunus serrulata var. spontanea, and Pinus densiflora. The results of the study are as follows : 1. $SO{_4}^{-2}$, $NO{_3}^-$ and $Cl^-$ concentrations are higher in the soils of the urban area than in those of the rural area, and $NO{_3}^-$ and $SO{_4}^{-2}$ are higher in the leaves than in the roots due to the absorption of the these pollutants through the stomata. 2. Ginkgo biloba, Robinia pseudoacacia. Zekova serrata, Quercus acutissima, and Platanus occidentalis can be adequated to the roadside trees and the environmental trees due to their good absorption of $NO{_3}^-$ and $SO{_4}^{-2}$. 3. Heavy metals in the soils of both areas are in the order of Mn > Zn > V > Cr > Pb > Ni > Cu > Mo> Cd, and in the leaves and roots of the trees in the both areas are in the order of Mn>Zn>Cr>Cu>V>Ni. Both orders are similar ones except V. There are more in the urban soils than in the rural soils in amount of Mn, Zn, Pb, V, Cu. 4. It is supposed that there is no antagonism between Mn and Zn in this study. 5. Se, Co and As are not detected in the soils, the leaves and the roots in both areas. Sn, Mo, Cd and Pb are also not detected in the leaves and roots in spite of considerable amount in the soils of both areas.

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Comparison of Nutrient Intakes of Korean Elderly People Living in Rural Area Between 24-hour Recall and Food Frequency Method (24시간회상법과 식품섭취빈도조사법을 이용한 농촌지역 노인의 영양소 섭취수준 비교)

  • Cho, Young-Eun;Lee, Soo-Lim;Cho, Eun-Hyun;Lomeda, Ria-Ann L;Kwak, Eun-Hee;Kim, Yang-Ha;Kwun, In-Sook
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.35 no.6
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    • pp.698-707
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    • 2006
  • Precise and correct assessment of nutrient intake using appropriate dietary intake method is necessary to improve the nutritional status of the target people group. It is more likely so in elderly people since their dietary habits and inadequate nutrient intakes are of special concern even worse in rural areas. Two popular methods of gathering dietary intake are 24-hour recall and food frequency method (or food frequency questionnaire). In the present study, we conducted both 24-hour recall and food frequency method on 120 rural elderly people $(aged\;65{\sim}74\;years)$, 62 male and 58 female. The dietary intakes were analyzed and the correlation of the dietary intake between 24-hour recall and food frequency method to determine the reliability between two methods. Also each nutrient intake was compared to recommended dietary allowance (RDA) or nutrient reference value (NRV) as appropriate. Even correlation did not show significance between the two methods except for dietary fiber intake (0.65 at p<0.01) and energy-yielding nutrients which mostly showed similar results with nutrient intake level. These support the possible reliability between 24-hours record and food frequency method that is also shown on the cases of most mineral and vitamin intakes. Thus, the food frequency method used in this study for the assessment of nutrient intakes of the elderly people would be reliable for the assessment of the nutrient intake along with the 24-hour recall. Also, the present study results suggested that rural female elderly people would be under optimal nutriture status due to the lower food intake measured by 24-hour recall. The optimal food frequency method would be considered to be developed for the precise nutrient assessment.

Studies on the Viability of Frozen Removed Seminal Plasma by Saline(RSP-S) and Tris-buffer(RSP-T) Semen of Small Spcies Dogs (소형 개 RSP-S와 RSP-T 정액의 동결 융해후의 생존성에 관한 연구)

  • 김상근
    • Korean Journal of Animal Reproduction
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    • v.25 no.3
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    • pp.269-275
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    • 2001
  • This study was carried out to investigate the general characteristics such as volume, sperm concentration, sperm motility, sperm abnormality on whole semen, RSP-S and RSP-T semen and fractional semen of small size dogs, and the effect of temperature and preservation time and cryoproservation on motility of whole and RSP-S and RSP-T semen. Multiple ejaculates were collected from small dogs by the digital manipulation of penis. 1. The volume per ejaculate semen, sperm of concentration and motility and abnormal sperm rate of 1st fractional semen were 0.65$\pm$0.09 $m\ell$, 4.52$\pm$0.35$\times$10$^{6}$ cells/$m\ell$, 15.64$\pm$3.85% and 5.50$\pm$0.62%. Also, 2nd fractional semen were 1.25$\pm$0.20$m\ell$, 3.35$\pm$0.48$\times$10$^{6}$ cells/$m\ell$, 96.25$\pm$4.65% and 4.24$\pm$0.46%. And 3rd fractional semen were 1.45$\pm$0.21$m\ell$, 3.55$\pm$0.52$\times$10$^{6}$ cells/$m\ell$, 92.82$\pm$4.24% and 4.66 $\pm$0.58%, respectively. 2. The sperm of concentration and motility and abnormal sperm rates of whole, RSP-S and RSP-T semen were 5.45$\pm$0.82$\times$10$^{6}$ cells/$m\ell$, 95.55 $\pm$4.65%, 4.58$\pm$0.45% and 4.82$\pm$0.36$\times$10$^{6}$ cells/$m\ell$, 90.10$\pm$3.42%, 6.48$\pm$0.68% and 4.55$\pm$0.45$\times$10$^{6}$ cells/$m\ell$, 93.25$\pm$3.85%, 4.82$\pm$0.58%, respectively. 3. The motility of whole, RSP-S and RSP-T semen were higher at 4$^{\circ}C$ than at 38$^{\circ}C$. When preservation temperature was at 4$^{\circ}C$, survival rates of RSP-S and RSP-T sperm were 97.54~6.25% at 1~72 hrs, 97.40~5.62% at 1~100 hrs, respectively. 4. The survival rates of slow and rapid frozen 2nd fraction, RSP-S and RSP-T semen were 67.3$\pm$4.45%, 88.8$\pm$4.46% and 46.4$\pm$3.84%, 74.4$\pm$4.20%, respectively. Survival rates was significantly higher in frozen RSP-S and RSP-T semen than that in control group(8.5$\pm$2.12%).

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Effect of Cryoprotectant Kinds and Cell Stages on the Viability of Mouse Embryos Cryopreserved by OPP Vitrification (동결보호제의 종류 및 배발달단계가 OPP Vitrification 동결보존시 생쥐수정란의 생존성에 미치는 영향)

  • 공일근;조성균;조성근
    • Korean Journal of Animal Reproduction
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    • v.23 no.1
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    • pp.85-92
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    • 1999
  • This study was designed to determine effect of cryoprotectant kinds and cell stages on OPP vitrification method in mouse embryos. The freezing speed, cryoprotectants and cell stage could affect of embryo viability following various vitrification methods. The vitrification solution used were consisting of 40% (v/v) ethylene glycol, 18% (w/v) Ficoll, 0.3 M sucrose solution in holding medium (D-PBS supplemented with 5% FCS: HM) (EFS) or 16.5% ethylene glycol , 16.5% dimethyl sulfoxide, 0.5 M sucrose in HM (EDS). The embryos were collected from oviduct at 18 h after hCG injection and then washed and cultured in mHTF medium until use. In experiment 1, the blastocysts were vitrified by OPP straw to determine the optimal vitrification solution of EFS or EDS. The post-thaw survival rates at re-expanded stage rates were significantly different between EFS and EDS (95.0 vs 100%), but at hatching stage was not different between EFS and EDS (90.0 vs 95.0%). respectively. In experiment 2, zygotes, 2-, 4-cell, morula and blastocysts were vitrified by OPP method to determine the acceptable of early stage embryos. The development rates to expanded blastocyst in zygote (70.0%) were significantly lower rather than those in 2-, 4- 8-cell, compacted morula or blastocyst (89.7, 90.0, 92.8, 97.6 or 97.5%), respectively. However, the cell number of post-thaw developed to expanded blastocyst in blastocyst and control blastocyst stage (39.6$\pm$2.81, 35.7$\pm$2.98) were significanty higher than those in zygote, 2-, 4-, 8-cell, compacted morula (29.8$\pm$3.21, 31.3$\pm$3.83, 29.3$\pm$3.58, 28.9$\pm$3.21 or 30.8$\pm$2.93). In experiment 3, the zygotes were exposed in VSl for 1, 2, and 3 min to the optimal exposed time. The cleavage rates (91.6, 88.5, 88.9%) and develop mental rates to blastocyst (83.3, 74.3 and 69.4%) depends on the exposed time in VSl were not significantly different among 1, 2, or 3 min, respectively. The cell number also were not significantly different among exposed time in VS1. respectively. These results indicate that OPP method could be useful for vitrification either EFS or EDS vitrification solution. The post-thaw survival rates at zygote were significantly lower than those at 2-, 4-, 8-cell, morula or blastocyst, respectively. The zygote stage were more sensitive rather than late stage embryos. The exposing time in VS1 for 1 min was better than that for 2 or 3 min, even it was not significantly different. The OPP vitrification method could be useful of mouse embryos either with EFS or EDS vitrification solution.

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Effect on In­Vitro Fertilization of Pig Oocytes Matured in Different In­Vitro Maturation Media according to Sperm Concentration of Liquid Boar semen (돼지 액상정액의 정자 주입농도가 서로 다른 체외성숙배지에서 배양된 난포란의 체외수정에 미치는 영향)

  • 박창식;이영주;고현진;양창범;손동수;서길웅;이규승
    • Korean Journal of Animal Reproduction
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    • v.26 no.1
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    • pp.1-7
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    • 2002
  • The present study was carried out to investigate the effects of maturation media on penetrability of pig oocytes by liquid boar sperm coincubated with different sperm concentrations in a modified Tris­buffered medium (mTBM). Follicular oocytes collected from ovaries of prepubertal gilts were matured in a modified TCM-199 (mTCM-199) medium, modified Waymouth MB 752/1 (mWaymouth MB 752/1) medium or NCSU-23 medium. Oocytes (30~40) were transferred into each well of a Nunc 4-well multidish containing 0.5 $m\ell$ maturation medium. The sperm­ich portion of ejaculates with greater than 90% motile sperm were used in the experiment. The semen was cooled 22 to 24$^{\circ}C$ over 2 h period. The semen was diluted with Beltsville Thawing Solution (BTS) extender at room temperature to give 2$\times$10$^{8}$ sperm/$m\ell$ in 100 $m\ell$ plastic bottle. Liquid boar semen of 30 $m\ell$ in 100 $m\ell$ plastic bottle was kept at 17$^{\circ}C$ far 5 days. The sperm with greater than 70% motility after day 5 of storage were used for in-vitro fertilization (IVF). After 44 h maturation of immature oocytes in 5% $CO_2$in air at 38.5$^{\circ}C$, cumulus cells were removed and oocytes (30~40) were coincubated for 6 h in 0.5 $m\ell$ mTBM fertilization medium with five different (1$\times$10$^{6}$ , 2$\times$10$^{6}$ , 4$\times$10$^{6}$ , 6$\times$10$^{6}$, 10$\times$10$^{6}$ $m\ell$) sperm concentrations. At 6 h after IVF, oocytes were transferred into 0.5 $m\ell$ NCSU-23 culture medium fur further culture of 6 h. At 12 h after IVF, sperm penetration, polyspermy and male pronuclear formation of oocytes were evaluated. Oocytes of NCSU-23 maturation medium decreased polyspermy and increased male pronuclear formation compared to those of mTCM­199 and mWaymouth MB 752/1 maturation media. Of oocytes matured in NCSU-23 medium and inseminated in mTBM medium with 2~4$\times$10$^{6}$ $m\ell$ sperm concentrations, 50.8~50.9% showed sperm penetration, 13.3~19.5% polyspermy and 43.9~45.4% male pronuclear formation. In conclusion, we found out that oocytes matured in NCSU­23 medium and inseminated in mTBM medium showed superior in­vitro fertilization compared to those matured in mTCM­199 and mWaymouth MB 752/1 maturation media and inseminated in mTBM medium. The optimum sperm concentrations for in-vitro fertilization of oocytes matured in NCSU-23 medium by liquid boar semen stored at 17$^{\circ}C$ for 5 days were 2~4$\times$10$^{6}$ $m\ell$.

In Vitro Fertilization of Pig Oocytes Matured In­Vitro by liquid Boar Spermatozoa (체외성숙 돼지 난포란의 액상정액을 이용한 체외수정)

  • 박창식;이영주
    • Korean Journal of Animal Reproduction
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    • v.26 no.1
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    • pp.17-23
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    • 2002
  • The present study was carried out to investigate the effects of the maturation media such as a modified TCM-199 (mTCM-199) medium, modified Waymouth MB 752/1 (mWaymouth MB 752/1) medium or NCSU-23 medium on penetrability of pig oocytes by liquid boar sperm. Oocytes (30~40) were transferred into each well of a Nunc 4-well multidish containing 0.5 $m\ell$ maturation medium. When immature pig oocytes were cultured in mTCM-199, mWaymouth MB 752/1 and NCSU-23 maturation media for 44 h in 5% $CO_2$, in air at 38.5$^{\circ}C$, the germinal vesicle breakdown (CVBD) rates of the oocytes were 95.6, 94.1 and 94.9%, respectively, and the maturation rates (metaphase II) of oocytes were 92.5, 90.1 and 91.1%, respectively. No differences were observed among the maturation media. The spermrich portion of ejaculates with greater than 90% motile sperm were used in the experiment. The semen was cooled 22 to 24$^{\circ}C$ over 2 h period. The semen was diluted with Beltsville Thawing Solution (BTS) extender at room temperature to give 2$\times$10$^{8}$ sperm/$m\ell$ in 100 $m\ell$ plastic bottle. Liquid boar semen of 30 $m\ell$ in 100 $m\ell$ plastic bottle was kept at 17$^{\circ}C$ for 5 days. The sperm with greater than 70% motility after day 5 of storage were used for in-vitro fertilization (IVF). After 44 h maturation of immature oocytes, cumulus cells were removed and oocytes (30~40) coincubated far 6 h in 0.5 $m\ell$ mTCM-199 and mTBM fertilization media with 2$\times$1061$m\ell$ sperm concentration. At 6 h after IVF, oocytes were transferred into 0.5 $m\ell$ mTCM-199 and NCSU-23 culture media for further culture 6 or 42 h. Sperm penetration, polyspermy and male pronuclear formation of oocytes at 12 h after IVF, and developmental ability of oocytes at 48 h after IVF were evaluated. The oocytes in combination with NCSU-23 medium for maturation and mTBM medium for IVF increased male pronuclear formation (48.0%) compared to those in combination with mTCM-199 media for maturation and IVF, and mWaymouth MB 752il medium for maturation and mTCM-199 medium far IVF. The rates of cleaved embryos (2~4 cell stage) at 48 h after IVF were 24.1% in combination with mTCM-199 media for maturation, IVF and culture, 43.6% in combination with mWaymouth MB 75211 medium fur maturation and mTCM-199 media for IVF and culture, and 71.2% in combination with NCSU-23 medium for maturation, mTBM medium for IVF and NCSU-23 medium for culture. In conclusion, we found out the oocytes matured in vitro were fertilized by liquid boar sperm stored in BTS extender at 17$^{\circ}C$ for 5 days. We recommend the simple defined NCSU-23 medium for nuclear maturation, mTBM medium and liquid boar sperm for IVF, and NCSU-23 medium for embryo culture.

Radioprotective Effects of Granulocyte-Colony Stimulating Factor in the Jejunal Mucosa of Mouse (생쥐에서 과립구 집락형성인자(Granulocyte-Colony Stimulating Factor)의 공장점막에 대한 방사선 보호효과)

  • Ryu, Mi-Ryeong;Chung, Su-Mi;Kay, Chul-Seung;Kim, Yeon-Shil;Yoon, Sei-Chul
    • Radiation Oncology Journal
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    • v.19 no.1
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    • pp.45-52
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    • 2001
  • Purpose : Granulocyle-colony stimulating factor (G-CSF) has been widely used to treat neutropenia caused by chemotherapy or radiotherapy. The efficacy of recombinant human hematopoietic growth factors in improving oral mucositis after chemotherapy or radiotherapy has been recently demonstrated in some clinical studies. This study was designed to determine whether G-CSF can modify the radiation injury of the intestinal mucosa in mice. Materials and Methods : One hundred and five BALB/c mice weighing 20 grams were divided into nine subgroups including G-CSF alone group $(I:10\;{\mu}g/kg\;or\;II:100\;{\mu}g/kg)$, radiation alone group (7.5 or 12 Gy on the whole body), combination group with G-CSF and radiation (G-CSF I or II plus 7.5 Gy, G-CSF I or II plus 12 Gy), and control group. Radiation was administered with a 6 MV linear accelerator (Mevatron Siemens) with a dose rate of 3 Gy/min on day 0. G-CSF was injected subcutaneously for 3 days, once a day, from day -2 to day 0. Each group was sacrificed on the day 1, day 3, and day 7. The mucosal changes of jejunum were evaluated microscopically by crypt count per circumference, villi length, and histologic damage grading. Results : In both G-CSF I and II groups, crypt counts, villi length, and histologic damage scores were not significantly different from those of the control one (p>0.05). The 7.5 Gy and 12 Gy radiation alone groups showed significantly lower crypt counts and higher histologic damage scores compared with those of control one (p<0.05). The groups exposed to 7.5 Gy radiation plus G-CSF I or II showed significantly higher crypt counts and lower histologic damage scores on the day 3, and lower histologic damage scores on the day 7 compared with those of the 7.5 Gy radiation alone one (p<0.05). The 12 Gy radiation plus G-CSF I or II group did not show significant difference in crypt counts and histologic damage scores compared with those of the 12 Gy radiation alone one (p>0,05). Most of the mice in 12 Gy radiation with or without G-CSF group showed intestinal death within 5 days. Conclusion : These results suggest that G-CSF may protect the jejunal mucosa from the acute radiation damage following within the tolerable ranges of whole body irradiation in mice.

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