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Studies on the In Vitro Development of Cloned Embryos by Somatic Cell Nuclear Transfer in Korean Native Goats (재래산양의 체세포 핵이식에 의한 복제수정란의 체외발달에 관한 연구)

  • Park H. S.;Kim T. S.;Jung S. Y.;Lee Y. H.;Jung J. Y.
    • Journal of Embryo Transfer
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    • v.20 no.2
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    • pp.105-112
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    • 2005
  • The present study was conducted to examine some factors affecting in vitro development of oocytes from somatic cell nuclear transfer (SCNT) in Korean native goats. Recipient oocytes were surgically collected after superovulation by using CIDR and FSH, PMSG, hCG and estrous synchronization in Korean Native goats. For nuclear transfer, the fibroblasts from caprine ear cells and fetal fibroblasts were surgically harvested and were cultured in vitro until cell confluency in serum-starvation condition (TCM-199 + $0.5\%$ FBS) for 3 to 5 days. The zona pellucidae of matured oocytes were partially drilled by laser irradiation. A single somatic cell was individually transferred into each enucleated oocyte. The reconstructed oocytes were then electrically fused and activated. Activated NT embryos were cultured in mSOF medium supplemented with $0.8\%\;BSA\;6\~7\;day\;at\;39^{\circ}C,\;5\%\;CO_2,\;5\%\;O_2,\;90\%\;N_2$ in air. There were no significant difference in the number of embryos cleaved and 4-cell development between the fibroblast nuclei from mature ear cells and fetal cells, but the rate of 8-cell development was higher (P<0.05) in ear cells $(40.5\%)$ than in fetal cells $(55.5\%)$. However, the embryo development to morula or blastocyst was not significantly different between both the groups$(6.7\%\;vs\;16.0\%)$, respectively. The number of embryo cleaved $(79.0\%)$ were higher (P<0.05) in the oocytes activated with ionomycin+6-DMAP than in the oocytes activated electrically $(9.5\%)$. The development of fused embryos to morula or blastocyst was found $15.6\%$ in ionomycin+6-DMAP, but no morula or blastocysts were developed in electrical stimulation. The development rate of SCNT embryos to morula or blastocyst was love. (P<0.05) in SCNT embryos $(19.0\%\;vs\;0.0\%)$ than that in parthenotes $(66.1\%\;vs\;59.1\%)$. In the parthenotes, the cleavage rate and development to morula or blastocyst were significantly higher (P<0.05) as $86.8\%\;and\;50.0\%$ in ovulated oocytes than in follicular oocytes $(69.0\%\;vs\;23.6\%)$, respectively. These results suggest that some factors Including superovulation treatment, oocyte source, maturation of follicular oocytes, activation method and culture condition may affect in vitro developmental capability of embryos produced by somatic cell nuclear transfer in Korean Native goats, and the fusion rate be greatly low compared with other species.

In Vitro Development and Survival Following Cryopreservation of Bovine Embryos according to Ovary Transport Temperature (난소 수송 온도에 따른 소 체외 수정란의 발육 및 동결-융해 후의 생존성)

  • Cho S.R.;Choi S.H.;Kim H.J.;Choe C.Y.;Jin H.J.;Son D.S.
    • Journal of Embryo Transfer
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    • v.21 no.2
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    • pp.163-168
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    • 2006
  • The present study was carried out to investigate in vitro development and post-thawed survivability of bovine embryos according to different ovary transport temperatures. Bovine ovaries were collected at a local slaughterhouse and were transported at 4 different temperature categories to laboratory: $7{\sim}10^{\circ}C\;(T1),\;11{\sim}17^{\circ}C\;(T2),\;18{\sim}25^{\circ}C\;(T3)$ and above $26^{\circ}C$ (control group). The cumulus-oocyte-complexes aspirated from ovaries were in vitro matured, fertilized and cultured. The rates of maturation (to metaphase II), cleavage and development to blastocysts were compared among treatment groups. Furthermore, frozen-thawed blastocysts were in vitro cultured to compare the survivability among groups. The maturation rates in the T1, T2 and T3 groups ($60.0{\sim}68.2%$) were significantly lower than that in the control group (81.8%, p<0.05). The cleavage rates in the T1 and T2 groups (52.6 and 54.5%) were significantly lower than that in the control group (83.6%, p<0.05). However, there was no difference in the development rate to blastocysts among all groups ($27.9{\sim}33.0%$, p>0.05). The survivability of frozen-thawed embryos was significantly lower in the T1 group (46.2%) than those in the T2, T3 and control groups ($68.8{\sim}7.13%$, p<0.05). In conclusion, the results suggest that ovary transport temperature at $26^{\circ}C$ may be optimal for the better in vitro development and the survival of frozen-thawed embryos produced in vitro Furthermore, exposure of ovary to temperature below $10^{\circ}C$ during transport may significantly decrease both in vitro development and survivability of frozen-thawed blastocysts.

Studies on the Application of Cattle Slurry in Grassland III. Grazing preference as affected by application time of cattle slurry on pasture (혼파초지에서 액상구비 시용에 관한 연구 III. 방목이용 초지에서 액상구비 시용시기에 따른 가축기호성 비교)

  • Kim, Jae-Kyu;Seo, Sung
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.13 no.1
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    • pp.43-48
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    • 1993
  • This experiment was carried out to determine the effects of application time of cattle slurry on grazing pasture on the grass growth, dry matter yield, nutritive value, animal preference, and efficiency of pasture utilization. Four different application times of slurry(right now after grazing, 5th day, 10th day, and 15th day after grazing), and a no-slurry(control) plot were designed, 1991∼1992. Annual application amount of slurry was 40 ㎥/ha, and slurry was applicated in spring(2 times), summer and autumn. Total NPK fertilizer applicated were same as 280, 200 and 240 kg/ha in all treatment, respectively. The pasture was grazed with growing beef cattle(initial body wt.; ca. 250 kg) at every 30∼35 cm of plant height. The grass height, dry matter yield, crude protein yield, and other nutritive value were not different among treatments. However, the animal preference and efficiency of pasture utilization showed significant difference by application time of slurry. The highest grazing preference (1: the worst∼9: the best). and utilization efficiency (0∼100%) were observed by application time of right now after grazing (7.8, 76%), which was same to control(7.8, 77%), and followed by 5th day after grazing(7.3, 74%) and l0th day after grazing(6.9, 71%). But the preference and pasture utilization were greatly decreased at application time of cattle slurry of 15th day alter grazing(5.5, 60%). From the above results, the optimum application time of cattle slurry on grazing pasture to enhance animal preference, and efficiency of pasture utilization was within 5 days after grazing (less than 13.6 cm of plant height), and within 10 days after grazing (less than 18.5 cm of plant height) at least.

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Effects of Probiotic Extracts of Kimchi on Immune Function in NC/Nga Mice (김치 추출 프로바이오틱스 섭취가 아토피 동물모델 NC/Nga mice에서 면역 지표에 미치는 영향)

  • Lee, In-Hoe;Lee, Sun-Hee;Lee, In-Seok;Park, Yoo-Kyoung;Chung, Dae-Kyun;Choue, Ryo-Won
    • Korean Journal of Food Science and Technology
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    • v.40 no.1
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    • pp.82-87
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    • 2008
  • Atopic dermatitis (AD) is characterized by chronic relapsing inflammation and is associated with hyper-production of immunoglobulin E (IgE). Recent studies have suggested that one of the treatments to alleviate symptoms of AD could be a supplementation of probiotics, Lactobacillus, Rhamnosus, Bifidus, etc. The purpose of this study was to evaluate the effects of probiotics on immune parameters in NC/Nga mice treated with 1-chloro-2,4-dinitro-benzene (DNCB). To induce atopic dermatitis, DNCB was treated to the back of mice for 2 weeks. Then, NC/Nga mice were divided into the four experimental groups randomly. Probiotics fragment, probiotics with other complex (Lactobacillus rhamnosus GG, Bifidobacterium lactis Bb-12LbL, L. plantarum K8, L. plantarum K8 fragment, ${\gamma}$-linolenic acid), antihistamine, and distilled water were administrated orally to the NC/Nga mouse for 4 weeks of experimental period. The groups were probiotics fragment group (DPF), probiotics with other complex group (DPOC), antihistamine group (DAH) and distilled water group (DDW) as a control group. The levels of serum IgE, interlukin-4 (IL-4), interlukin-5 (IL-5), interferon-gamma (IFN-${\gamma}$) and spleenocyte IgE were measured. The levels of serum IgE were significantly different among the four experimental groups. Before the treatment, there was no differences among the groups. However, from the first through the third week of the treatments, the levels of serum IgE in the probiotics (DPF, DPOC) and antihistamine (DAH) groups were lower than those of control group (p < 0.05). The levels of serum IL-4 of DPOC group was significantly lower than that of control group (p < 0.05) and serum IL-5 levels of DPF, DPOC, and DAH groups were significantly lower than that of control group. The levels of serum IFN-${\gamma}$ were not different among the four experimental groups. The levels of serum IgE in supernatant of spleen lymphocytes were not significantly different among the groups. These results suggest that probiotics supplementation showed partial effectiveness in the DNCB treated NC/Nga mice via modulation of IgE level and IL-4, IL-5 production. Based on these findings, probiotics exhibited the inhibitory effect via IL-4 production thereby inhibited the production of IgE in atopic animal model NC/Nga mice.

Current Situation on Signing Advance Medical Directives and Actual Life-sustaining Treatment Given at a University Hospital (일개 대학병원의 연명치료 선택 및 사전의료의향서 작성 현황)

  • Yoon, Ho-Min;Choi, Youn-Seon;Hyun, Jong-Jin
    • Journal of Hospice and Palliative Care
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    • v.14 no.2
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    • pp.91-100
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    • 2011
  • Purpose: This study was performed to investigate patients' preferences on receiving life-sustaining treatments (LST) and to analyze the relationship between patients' characteristics and LST selection. We also examined any discrepancy between LST patients' choices regarding medical intervention and actual medical intervention given/not given within 48 hours before death. Methods: This cross-sectional study was performed from March 1, 2008 to August 31, 2008 in the Palliative Care Unit of Korea University Hospital. Electric medical records (EMR) of 102 hospice cancer patients were reviewed, and 74 patients with Glasgow coma scale (GCS) ${\geq}$10 at the time of signing the advance medical directives (AMD) were selected for the first analysis. Then, patients alive at the end of this study, transferred to other hospitals or dead within 48 hours were excluded, and the remaining 42 patients were selected for the second analysis. Results: Preferred LST included antibiotics, total parenteral nutrition, tube feeding, transfusion, and laboratory and imaging studies. The relationship between patients' characteristics and LST could not be analyzed due to skewed preferences. LST chosen at the time of signing the AMD and actual medical intervention given/not given in the last 48 hours showed discrepancy in most cases. Conclusion: When making AMD in hospice cancer patients, it is important to consider the time and possibility of changing the choices. Above all, patients must fully understand the AMD. Thus, LST should always be provided with careful consideration of all possibilities, because legal and social aspects of AMD have not been established yet.

Proper Light Intensity, Potting Media, and Fertilization Level for Potted Orostachys iwarenge for. magnus (울릉연화바위솔 분화재배를 위한 적정 광도, 분용토 및 시비 수준)

  • Jeong, Kyeong-Jin;Chon, Young-Shin;Choi, Kyeong-Ok;Ha, Su-Hyeon;Yun, Jae-Gill
    • Horticultural Science & Technology
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    • v.30 no.4
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    • pp.357-362
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    • 2012
  • Proper light intensity, shading tolerance, potting media, and fertilization level were investigated to develop Orostachys iwarenge for. magnus (Korean name, Ullungyeonwhabawisol) as a potted ornamental plant. The plants were grown under different light intensity (52, 82, 90, and 97% shading). The best growth was shown at 52% shading, which indicated that the proper light intensity for O. iwarenge for. magnus is less than 52% of shading. Plant growth decreased severely at 82% shading and leaf color became lighter as the shading rate increased, which indicated that O. iwarenge for. magnus has no tolerance against low light intensity. To select a proper potting media, decomposed granite (DG), fertilizer-amended media (FAM), river sand (RS) were used as potting medium with different ratio of 60:20:20 (DG:FAM:RS, v/v/v), 80:20 (DG:FAM, v/v), 60:40 (DG:FAM, v/v), and 20:80 (FAM:RS, v/v). DG:FAM:RS (60:20:20) showed the highest values in shoot fresh weight, plant width, and number of runner in potted O. iwarenge for. magnus. Fresh weight of shoot part was 16 g in DG:FAM:RS (60:20:20), which was about 2 folds of those at the other medium. At the experiment for selection of proper fertilization level, plants showed a better growth as the concentration of hyponex solution and application frequency increased. Once drenching 1 week interval of hyponex solution diluted by 1,000 folds brought the highest results in fresh weight, plant width, and runner number. Particularly, fresh weight of shoot part was 35 g at once drenching per week of 1,000 folds solution, indicating 84% improvement comparing with non treatment (19 g).

A Study on Hygiene and Safety of Sanitary Wet Towel (물수건의 위생실태 및 안전성 연구)

  • Kim, Young-Sug;Kim, Yang-Hee;Kim, Young-Su;Kim, Dae-Hwan;Ryu, Kyong-Shin;Yoon, Mi-Hye
    • Journal of Food Hygiene and Safety
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    • v.31 no.4
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    • pp.258-263
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    • 2016
  • The risks of sanitary indicative bacteria, heavy metals and chlorinated derivatives in 94 cases of sanitary wet towels used in food services (39 from sanitary wet towel treatment business, 55 from food services) were assessed in the present study. Lead was detected in the range of N.D.~0.41 mg/kg (75 cases were not detected), N.D.~0.25 mg/kg of arsenic (93 cases were not detected), N.D.~0.01 mg/kg of cadmium (7 cases were lower than limit of quantitation; 87 cases were not detected), 0.003 mg/kg ~ 0.09 mg/kg of mercury. And chromium (VI) was not detected from all samples. The level of lead was the highest among the tested heavy metals, and the highest concentration of lead was 0.41 mg/kg. However, it was only 2.1% of legal limit (less than 20 mg/kg). The average moisture content of the samples was 61.9% (50.0% ~ 77.0%) and it showed no relevance to the detection of bacterial counts. Escherichia coli was not detected. Bacterial counts were detected 43 cases and among them, 24 cases were exceeded the legal limit. It was verified that the packaging conditions of sanitary wet towel (whether it is packed by a piece or not and sealed or not) are critical factors to cause the germ contamination and cross contamination in the wet towels. The chlorinated derivatives (chlorites and chlorates) were detected in 17 (19.3%) out of 88 cases. The results would be used as preliminary information to establish the programs of "Safety education for manufacturers and public policy of safety".

Effects of Whey Protein Hydrolyzates Fractionated by Molecular Weight on the Growth of Bifidobacterium bifidum Bb-11 (분자량에 따라 분획된 유청단백분해물이 Bifidobacterium bifidum Bb-11의 생장에 미치는 영향)

  • 김완섭;박승용;이범진;김평현;고준수
    • Food Science of Animal Resources
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    • v.22 no.1
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    • pp.59-65
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    • 2002
  • This study was carried out to evaluate the effect of whey protein concentrate-80%(WPC-80) and whey protein isolate(WPI) on the growth of B. bifidum Bb-11. Whey proteins($\alpha$-lactalbumin, $\beta$-lactoglobulin) were digested with trypsin, then their hydrolyzates were separated into three fractions (>10,000Da, 3,000∼10,000Da, <3,000Da) by two-step ultrafiltration process with Centriprep 10 and Centricon-30. These three fractions by molecular weight were evaluate growth-promoting effects for the B. bifidum Bb-11. The results obtained were summarized as follows; The growth rate of B. bifidum Bb-11 tended to increase by supplementation of WPC-80 to basal medium, but decreased by supplementation of WPI. Two whey proteins were hydrolyzed by trypsin at 40$\^{C}$ for 6 hrs, and three fractions were collected by UF treatment and concentrated by Centricon-30. Collected concentrations of protein of F-I and F-II and F-III from $\alpha$-lactalbumin were 11.53mg, 7.79mg, and 5.21 mg and those of protein from $\beta$-lactoglobulin were 4.13mg, 5.30mg, and 9.351mg, respectively. Three fractions of $\alpha$-lactalbumin hydrolyzates promoted the growth rate of B. dbifidum Bb-11. Growth promoting activities of hydrolyzates(F-I and F-II) with molecular weight below 10,000Da were stronger than that of hydrolyzate(F-III) above 10,000Da. However, there was no significant difference between the hydrolyzate F-I and F-II. Three fractions of $\beta$-lactoglobulin hydrolyzates improves the growth rate of B.bifidum Bb-ll. The growth of B.bifidum Bb-ll was decreased after 24 hr incubation by supplementation of either F-II or F-III fraction compared to basal Whey medium, but maintained the enhancement by supplementation of F-I.

Effects of Milk Production, Postparient Days or Seasons on In Vivo Embryo Production by Superovulation in Holstein Cows (유우의 과배란 처리에 있어서 산유량, 분만 후 처리시기 및 계절이 체내수정란 생산에 미치는 영향)

  • Lim, Kwang-Taek
    • Journal of Embryo Transfer
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    • v.24 no.1
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    • pp.33-37
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    • 2009
  • Multiple ovulation and embryo transfer (MOET) has the potential to increase the rates of genetic improvement in cattle. Thus this study was performed to investigate several factors influencing in vivo embryo production in Holstein cattle under field conditions. The donors were superovulated with Folltropin-V and $PGF_2{\alpha}$ combination method. From Day 10 onward, donors were superovulated by i.m., twice daily, administration of 400mg Folltropin-V given in a series of decreasing doses over a 4-day period: on the first day, 3.5ml; on the second day, 3.0ml; on the third day, 2.0ml; and on the fourth day, 1.5ml (20ml in total, equivalent to 400mg of NIH-FSH-P1). Estrus was induced by i.m. administration of 25mg prostaglandin $F_2{\alpha}$ on the sixth and seventh of FSH treatment. Estrus detection was performed twice daily beginning 24h after the first prostaglandin $F_2{\alpha}$ injection. Donor cows were artificially inseminated 12 and 24 h after first standing estrus with semen from a proven Holstein sire. Embryos used in this study were recovered Day 7.5 of the cycle (Day 0: first standing estrus). From 195 superovulated dairy cows, 2,104 eggs were recovered, of which 1,172 were classified as transferable embryos based on morphological evaluation of quality. The results are summarized as follows: 1. The numbers of recovered and transferable embryos did not significantly differ among the capacity of milk production that were < 10,000kg/305days (group 1), $10,000{\sim}12,000\;kg$/305days (group 2) or > 12,000kg/305 days (group 3) (p>0.05, Table 1). 2. No differences in the numbers of recovered and transferable embryos were found among the donor's postparient days (p>0.05, Table 2). 3. Also, the numbers of recovered and transferable embryos of each superovulation seasons did not significantly differ among the four groups (p>0.05, Table 3).

Relativeness between Growth and Bio-informations of Aeroponically Grown Tomato as Influenced by Spray Intervals of Nutrient Solution (양액의 분무간격에 따른 분무경재배 토마토의 생장 및 생체정보와의 관련성)

  • 정순주;소원온;지전영남;영목방부
    • Journal of Bio-Environment Control
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    • v.1 no.2
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    • pp.154-161
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    • 1992
  • This experiment was carried oui to determine the relativeness between growth, yield characters and bio-informations as influenced by the spray and rest time intervals of nutrient solution. Tomato(Lycopersicon esculentum Mill.) were grown in aeroponic system on a misting schedule of continuously 60 sec, 30 sec and 10 sec at 10 min intervals with full strength Yamazaki's solution recommended for tomato production. The results obtained were as follows : 1. Leaf area was highest in the plot of 30 sec spray and 10 min rest while the forest one was the plot of 60 sec spray and 10 min rest. Growth characteristics in terms of dry weight of each organ, number of flower, number of flower setted and fruit dry weight were greater in the plot of 30 sec spray and 10 min rest than the other treatments. 2. The number of flower increased with decreasing dry weight but number of flower sorted was not significantly different among treatment except for the plot of 60 sec spray and 10 min rest. 3. Leaf dry weight and fruit dry weight were highly correlated so that 30 sec spray and 10 min rest plot which is the highest fruit dry weight showed the largest leaf area. Continuously sprayed plot reduced markedly the fruit dry weight compared with leaf area. Optimum spray and rest time of nutrient solution in the range of this experiment was determined as 30 sec spray and 10 min rest. 4. Solar radiation within glasshouse during daytime reduced severely compared with outdoor one and air temperature within greenhouse was higher than the leaf temperature of tomato plant. The changes of environmental factors, solar radiation, temperature were accompanied with the sensitive change of bio-informations of tomato leaf Especially differences of spray intervals of nutrient solution affected greatly to the changes of bio-informations : leaf water potential, stomatal resistance and leaf temperature etc. 5. The changing patterns of leaf growth as influenced by the spray and rest intervals of nutrient solution were closely related to the leaf water potential, stomatal resistance and leaf temperature. Feasibility was demonstrated that measurement of bio-information of tomato leaf as influenced by the change of environmental factors could be expected to the amount of growth and fruit yield.

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