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Development of Transgenic NT Embryos Using Bovine Fetal Fibroblasts Transfected with hFSH Gene (hFSH 유전자가 도입된 소 태아섬유아세포를 이용한 형질 전환 복제 수정란의 발달)

  • Yang B.C.;Im G.S.;Kim D.H.;Min K.S.;Yoon D.H.;Park H.S.;Kim S.W.;Hwang I.S.;Seo J.S.;Seong H.H.;Yang B.S.
    • Journal of Embryo Transfer
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    • v.21 no.1
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    • pp.13-20
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    • 2006
  • The purpose of this study was to develope the transgenic cattle expressing hFSH into the urine using the nuclear transfer. To produce the interest gene in urine, the specific vector was ligated with hFSH gene undo. maUII promoter. The fetal fibroblast cells (KbFF) were isolated from a 45-day male fetus. The hFSH gene was co-transfected with pcDNA3 (neo) vector to KbFF cells by electroporation. The gene-transfected cells were cultured with G-418 selection medium for 2 weeks. Selected colonies were confirmed by PCR. For nuclear transfer, enucleated bovine oocytes were transferred with hFSH transfected or nontransfected fetal fibroblasts. The cleavage and blastocyst formation rates were significantly lower (p<0.05) in cloned embryos transfected with hFSH gene (68.7% and 15.7%) than in those non-transfected (67.6% and 24.5 %), respectively. Apoptosis analysis showed no difference between hFSH transfected and non-transfected blastocysts (p>0.05). The blastocysts were transfected to 77 (control 24, hFSH 53) recipient cows. Two calves were born (1.9%) following transfer with NT embryos transfected with hFSH gene, but they were confirmed not to be transgenic calves. This result shows that the hFSH colonies were mixed with transfected and non transfected cells. Further research will be needed for selection and establishment of gene transfected cells.

Characteristics and Immunomodulating Activity of Lactic Acid Bacteria for the Potential Probiotics (Probiotics로서의 젖산균주의 특성 및 면역활성)

  • Seo, Jae-Hoon;Lee, Ho
    • Korean Journal of Food Science and Technology
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    • v.39 no.6
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    • pp.681-687
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    • 2007
  • This study was designed to examine the suitable characteristics of potential probiotic bacteria. Possible probiotic bacteria, including Lactobacillus acidophilus DDS-1, Lb. acidophilus B-3208, Bifidobacterium bifidum KCTC 3357, Lb. plantarum, Leuconostoc mesenteroides ssp. mesenteroides ATCC 8293, and Lactococcus lactis ssp. lactis ATCC 7962 were selected. We then measured their acid and bile tolerances, adhesion properties in the gastrointestinal tract, antimicrobial activity against pathogenic bacteria, and immunomodulation activity. The acid tolerances of Lb. acidophilus DDS-1, Lb. acidophilus B-3208, Lb. plantarum, and Leu. mesenteroides ssp. mesenteroides ATCC 8293, in PBS (pH 2.5) for 2 hr, were high enough that 50% of the inocula survived. The bile tolerances of all bacteria, except Lc. lactis ssp. lactis ATCC 7962, were also observed at a 3% oxgall concentration in MRS broth. The results of the adhesion property assay showed that the total binding affinities of Lb. acidophilus DDS-1, Lb. acidophilus B-3208, and B. bifidum were about three times higher than those of the other bacteria. In testing their antimicrobial activities against pathogens, Lb. acidophilus B-3208, B. bifidum KCTC 3357, and Lb. plantarum inhibited the growth of pathogenic bacteria. For their immunomodulation activity, the cell wall fractions from Lb. acidophilus DDS-1 and Lb. acidophilus B-3208 showed the highest bone marrow cell proliferation activities. However, the cell wall fractions of Lb. acidophilus DDS-1 and B. bifidum, and the cytosol fraction of Lc. lactis ssp. lactis ATCC 7962 showed higher macrophage stimulation activities than those of the other bacteria. Since Lb. acidophilus DDS-1 and Lb. acidophilus B-3208 satisfy the requirements for probiotics, they can be considered suitable probiotic bacteria.

Quality Changes of a Fully Ripe Korean Native Pumpkin, Yangsan, during Long-term Storage, and High Temperature and Pressure Treatment (장기저장 및 고온고압 처리에 따른 한국재래종 호박 '양산'의 품질변화)

  • Youn, Sun-Joo;Jeong, Byeong-Ryong;Kang, Sun-Chul
    • Applied Biological Chemistry
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    • v.47 no.4
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    • pp.409-413
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    • 2004
  • We have studied quality changes of fully ripe fruit of Korean native pumpkin 'Yangsan' regarding the following parameters: pH, sugar content, weight, water content, contents of crude protein and amino acids during 60 days storage at room temperature. As the results, there was no changes in sugar contents according to the storage period, but pH was changing to a little acidic direction with slight decrease in weight and water content. Contents of total crude proteins and comprising amino acids were increased during the storage period. The main contents of amino acids of the Korean native pumpkin, Yangsan, were glutamic acid (15.5%), aspartic acid (10.1%), lysine (8.7%), valine (7.5%), leucine (7.1%) and alanine (6.6%), which were not highly influenced during storage period. Additionally we have investigated the content of free amino acids and color changes during processing of Yangsan under high temperature at $121^{\circ}C$ and high pressure at $1\;kg/cm^2$. In fully ripe fruits, a total of 29 kinds of free amino acids were detected including 8 kinds of essential amino acids (histidine, isoleucine, leucine, lysine, phenylalanine, methionine, threonine and valine). More than 35% of total free amino acids were aspartic acid (20.3%) and asparagine (15.4%); ornithine, citrullin, and arginine, which are related to Ornithine cycle, were also detected in fully ripe fruits. But when treated with high temperature and high pressure, glutamic acid and arginine were decreased rapidly whereas ammnonium chloride was relatively increased. Moreover "b" value as yellow color indicator was decreased from 17.45 to 9.14 while treated for 60 minutes with high temperature and pressure, caused by the degradation of ${\beta}-carotene$ and other yellowish pigments in Yangsan.

Effects of Feeding Levels and Particle Size of Germanium Biotite on Pig Performance (돼지 생산성에 있어 게르마늄흑운모의 첨가수준 및 입자도의 효과)

  • Lee, W.B.;Kim, I.H.;Hong, J.W.;Kwon, O.S.;Min, B.J.;Shon, K.S.;Jung, Y.K.
    • Journal of Animal Science and Technology
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    • v.45 no.5
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    • pp.787-796
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    • 2003
  • Two experiments were conducted to investigate the effect of germanium biotite (GB) on growth performance, nutrient digestibility and fecal gas emission in pigs. In Exp 1., a total of one hundred nursery pigs (initial body weight 13.12${\pm}$0.15kg) were used in a 21 d growth assay. The five treatments were control (CON, basal diet), GB0.5-200 (basal diet + 0.5% GB, 200mesh), GB1.0-200 (basal diet + 1.0% GB, 200mesh), GB0.5-325 (basal diet + 0.5% GB, 325mesh), GB1.0-325 (basal diet + 1.0% GB, 325mesh). Each treatment had four replicates with five pigs per replicate. ADG, ADFI and gain/feed were not significantly different among the treatments. Fecal NH3-N concentration of pigs fed the GB325 diet was lower than that of pigs fed the GB200 diet (P=0.01). The GB treatments reduced fecal volatile fatty acids significantly compared to the CON (propionic acid, P=0.01; butyric acid, P=0.01; acetic acid, P=0.02). Especially, fecal propionic acid concentration of pigs fed the GB325 diets was lower than that of pigs fed the GB200 diets (P=0.02). In Exp 2., a total of seventy five pigs (initial body weight 21.18${\pm}$0.15kg) were used in a 28 d growth assay. The treatments were same as described for Exp. 1. Each treatment had five replicates with three pigs per replicate. The GB1.0 treatments significantly increased the ADG compared to the GB0.5 treatments (P=0.03). The DM and N digestibility of pigs fed the GB1.0 diets were higher than that for pigs fed the GB0.5 diets (P=0.01). Also, the Ca digestibility of pigs fed the GB diets was higher than that for pigs fed the CON diets (P=0.01). The fecal NH3-N concentrations for the GB treatments were lower than that for the CON (P=0.01). The GB325 treatments significantly decreased the fecal NH3-N concentration compared to the GB200 treatments (P=0.03). The fecal butyric acid concentration for the GB325 treatments was lower than that for the GB200 treatment (P=0.04). In conclusion, the results obtained from these feeding trials suggest that the dietary GB for nursery pigs affects fecal noxious gas emission. In growing pigs, dietary GB was effective to improve ADG and decrease fecal noxious gas emission.

Cold Tolerance of Ground Cover Plants for Use as Green Roofs and Walls (옥상 및 벽면녹화용 지피식물의 내한성 비교)

  • Ryu, Ju Hyun;Lee, Hyo Beom;Kim, Cheol Min;Jung, Hyun Hwan;Kim, Ki Sun
    • Horticultural Science & Technology
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    • v.32 no.5
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    • pp.590-599
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    • 2014
  • This study was conducted to compare the cold tolerance of seven ground cover species, Orostachys japonica, Sedum oryzifolium, S. kamtschaticum 'SG1', S. reflexum, S. rupestre 'Blue Spruce', S. spurium 'Green Mental', and S. takesimense, which have been used for green roof and wall systems in Korea. Plants were grown in 10-cm pots and 1 g of tissues at stem-end and crown of each species were kept under either light or dark condition, respectively. For cold tolerance tests, plants were initially left at $4^{\circ}C$ and linearly cooled to 0, -4, -8, -12, -16, and $-20^{\circ}C$ at $-2^{\circ}C{\cdot}h^{-1}$ rate. Low temperature injury and regrowth rates were visually evaluated and assessed by image analysis, respectively. The lethal temperature ($LT_{50}$) of plant species was determined using electrolyte leakage measurements. S. reflexum was the most cold tolerant, showing the most survival at $-16^{\circ}C$, whereas S. oryzifolium and S. takesimense showed low temperature injury at $-8^{\circ}C$. Similar results were found with electrolyte leakage measurements at the stem end. For each species, the crown (Mean $LT_{50}:\;-12.15^{\circ}C$) was more cold tolerant than the stem end (Mean $LT_{50}:\;-10.47^{\circ}C$). In conclusion, S. reflexum and S. rupestre 'Blue Spruce' are recommended for planting in the central region of Korea during late fall and early winter, as they were more cold tolerant and showed more vigorous regrowth than the other tested plant species.

Evaluation of Macrophage Activity and Repeated Oral Dose Toxicity in Sprague-Dawley Rats on Multivitamin (종합비타민의 랫드에서 반복투여독성 시험과 대식세포 기능 활성 평가)

  • Kim, Hye-Ri;Jang, Hye-Yeon;Lee, Hae-Nim;Park, Young-Seok;Park, Byung-Kwon;Kim, Byeong-Soo;Kim, Sang-Ki;Cho, Sung-Dae;Nam, Jeong-Seok;Choi, Chang-Sun;Chang, Soon-Hyuk;Jung, Ji-Youn
    • Journal of Food Hygiene and Safety
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    • v.28 no.4
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    • pp.360-366
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    • 2013
  • The objective of this study is to investigate the effect of multivitamin on macrophage activity in Raw 264.7 cell and repeated oral dose toxicity in Sprague-Dawely rat of multivitamin. Raw 264.7 cells were treated with 50 and $100{\mu}g/mL$ multivitamin for 24 h. To measure the activity of macrophages, NO and TNF-${\alpha}$ assays were performed in Raw 264.7 cells. Treatment with 50 and $100{\mu}g/mL$ multivitamin for 24 h significantly increased production of NO and TNF-${\alpha}$ compared with control groups, indicating activation of macrophages. The female rats were treated with multivitamin of control group, low group (0.24 g/kg), medium group (1 g/kg) and high group (2 g/kg) intragastrically for 4 weeks, respectively. We examined the body weight, the feed intake, the clinical signs and serum biochemical analysis. We also observed the histopathological changes of liver, ovary, brain, adrenal gland, spleen, kidney, heart and lung in rats. No significant differences in body weights, feed intake, biochemical analysis and histopathological observations between control and multivitamin treatment group were found. In conclusion, multivitamin is physiologically safe and improve macrophage activity.

Retrograde Tracer Studies of Tecto-Reticulospinal Pathway and Dorsal Lateral Geniculate Nucleus on GluR1- and GluR4-Immunoreactive Neurons in the Hamster Superior Colliculus (Tecto-reticulospinal pathway (TRS)와 dorsal lateral geniculate nucleus (dLGN)에서 역행성이동추적물질 이용 햄스터 상구에서 GluR1-, GluR4- 면역반응 신경세포 연구)

  • Choi, Jae-Sik;Lee, Jea-Young;Jang, Yu-Jin;Lee, Eun-Shil;Jeon, Chang-Jin
    • Journal of Life Science
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    • v.20 no.1
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    • pp.1-8
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    • 2010
  • We recently reported the distributions of AMPA ($\alpha$-amino-3-hydroxyl-5-methyl-4-isoxazole-propionate) receptor subtypes glutamate receptors (GluR) 1 and GluR4 in the superior colliculi (SC) of hamsters with antibody immunocytochemistry and the effect of enucleation on these distributions. We also compared these labelings to those of calcium-binding proteins calbindin D28K, calretinin, parvalbumin, and GABA. In the present study, we investigated whether the GluR1- and GluR4-immunoreactive (IR) neurons are interneurons or projection neurons by injection of the retrograde tracer horseradish peroxidase (HRP) into one of each major ascending and descending pathways of the SC. HRP injections were made into a tecto-reticulospinal pathway (TRS) and dorsal lateral geniculate nucleus (dLGN). Animals were then allowed to recover and to survive for 48 hr before perfusion. Sections containing retrograde-labeled neurons were then treated for GluR-immunoreactivity. HRP injections proved that only a small population of the GluR1-IR cells project into TRS (1.4%) and dLGN (2.6%). However, a large subpopulation of GluR4-IR cells project into TRS (32.7%). The differential compositions of inter/projection neurons, along with our previous studies on the separate distribution of the GluR subunits, its differential co-localization with calcium-binding proteins and GABA, and differential reactions to enucleations, strongly imply the functional variety of the receptor subunits in visual behavior responses.

Detoxicating Effects of Oriental Herb Extract Mixtures on Nicotine and Dioxin (생약재 추출물의 nicotine 및 dioxin 해독효과)

  • Park, Ki-Moon;Hwang, Jin-Kook;Shin, Kyoung-Min;Kim, Hyun-Suck;Song, Jae-Hwan
    • Korean Journal of Food Science and Technology
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    • v.35 no.5
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    • pp.980-987
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    • 2003
  • In this report, we investigated the detoxication effects of Saururus chinenis, Geranium nepalense, Lonicera japonica, Cassia obtusifolia, Glycyrrhiza uralensis, or their mixtures by employing acute toxicity tests for nicotine and dioxin. When fatal doses $(LD_{100}\;=\;42\;mg/kg)$ of nicotine were injected into the abdominal cavities of ICR mice, those treated with OHEM showed delayed paralysis, half the duration of hyperactivity, and a 73 % survival rate. The results revealed the strong detoxicating effects of the mixtures. We also measured the amount of the degradation product of nicotine and cotinine in humans. Consumption of OHEM promoted (he more specific) the metabolic pathways of nicotine, increasing continine excretion by 1.5 times. As a result the amount of cotinine in urine was reduced to less than 5% after treatment with OHEM. In order to test the toxicity of dioxin, we used TcnN(SD)BR rats exposed to TCDD. While TCDD treatment reduced the blood levels of hemoglobin and platelet, OHEM consumption relieved these effects and, furthermore, helped to recover the number of platelet to the normal level (p<0.05). Moreover, neutrophils (%) and monocytes (%), which were reduced by the injection of TCDD, recovered to normal levels upon treatment with OHEM. The amount albumin reduced by TCDD (p<0.05) normalized, while the activities of GOT and GTP increased by TCDD were reduced. Increases in total cholesterol and neutral fatty acids induced by TCDD were also reduced by OHEM injection (p<0.05). In the kidney, TCDD-induced rises in creatinine were suppressed by OHEM treatment, while decreases in iron levels from TCDD were raised to normal. The treatment of TCDD had more toxic effects in the blood and pancreas than on the liver, kidney and heart. On the other hand, the detoxication of OHEM had significant effects on the liver and pancreas. The normalization by OHEM of various clinical abnormalities induced by TCDD demonstrates the detoxicating effect of OHEM that ameliorates systemic metabolism not properly functioning.

Antioxidative and Biological Activity of Hot Water and Ethanol Extracts from Phellinus linteus (상황버섯 열수 및 에탄올 추출물의 항산화 효과 및 생리활성)

  • Kim, Jung-Ok;Jung, Mee-Jung;Choi, Hyang-Ja;Lee, Jin-Tae;Lim, Ae-Kyoung;Hong, Joo-Heon;Kim, Dae-Ik
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.37 no.6
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    • pp.684-690
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    • 2008
  • To investigate the applicability of hot water extract (PLW) and ethanol extract (PLE) from Phellinus linteus as functional food and cosmeceutical materials, its total flavonoids content, total phenolics content, electron donating ability (EDA), nitrite-scavenging ability (NSA), SOD-like activity, inhibitory effect of tyrosinase and elastase were examined. Total flavonoids contents of PLW and PLE were 17.31 mg/g and 42.61 mg/g, respectively, and total phenolics contents were estimated as 149.92 mg/g for PLW and 432.42 mg/g for PLE. The EDA of PLW and PLE were $6.49{\sim}92.98%$ and $22.61{\sim}94.28%$. The EDA and total phenolics contents had a high correlation of 0.83. The NSA was pH dependent, and was highest at pH 1.2 and lowest at pH 6.0. The NSA of PLE was higher than that of PLW. The SOD-like activities of PLW and PLE were $14.36{\sim}35.21%$ and $17.27{\sim}81.84%$, respectively, and the activity was dependent on the sample concentration. The tyrosinase inhibitory activity was the highest in PLE ($10.51{\sim}80.93%$) while that of PLW was $4.77{\sim}43.69%$. Finally, the elastase inhibitory activity was $10.01{\sim}76.02%$ at PLE. Based on the above results, we deemed that the ethanol extract of Phellinus linteus was the most pertinent for use as functional food and cosmeceutical materials.

The Influences of Obstructive Apneas on Changes of Cardiovascular Function in Anesthetized Dogs with $\alpha$-chloralose ($\alpha$-chloralose로 마취한 개에서 폐쇄성 무호흡이 심혈관계 기능변화에 미치는 영향)

  • Jang, Jae-Soon;Kang, Ji-Ho;Lee, Sang-Haak;Choi, Young-Mee;Kwon, Soon-Seog;Kim, Young-Kyoon;Kim, Kwan-Hyoung;Song, Jeong-Sup;Park, Sung-Hak;Moon, Hwa-Sik
    • Tuberculosis and Respiratory Diseases
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    • v.48 no.3
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    • pp.347-356
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    • 2000
  • Background : Patients with obstructive sleep apnea syndrome are known to have high long-term mortality compared to healthy subjects because of their cardiovascular dysfunction. The observation of hemodynamic changes by obstructive apneas is helpful when attempting to understand the pathophysiological mechanism of the development of cardiovascular dysfunction in those patients. Therefore, we studied the changes in cardiovascular function with an animal model and tried to obtain the basic data for an ideal experimental model (this phrase is unclear), a requirement for a more advanced study. Methods : Sixteen anesthetized dogs with ${\alpha}$-chloralose delete were divided into two groups : 8 dogs of room air breathing group and 8 dogs of oxygen breathing group. We measured $PaO_2$, $PaCO_2$, heart rate, cardiac output, mean femoral artery pressure, and mean pulmonary artery pressure at specified times during the apnea-breathing cycle before endotracheal tube occlusion (baseline), 25 seconds after endotracheal tube occlusion (apneic period), 10 seconds (early phase of postapneic period, EPA) and 25 seconds (late phase of postapneic period, LPA) after spontaneous breathing. Results : In room air breathing group, the heart rate significantly decreased during the apneic period compared to that at baseline (P<0.01) and increased at EPA and LPA compared to that during the apneic period (P<0.01). But, the heart rate showed no significant changes during apneic and postapneic periods in the oxygen breathing group. Cardiac output tended to decrease during apneic period compared to that at baseline, but was statistically significant. Cardiac output significantly decreased at LP A compared to at baseline (P<0.01). Mean femoral artery pressure was significantly decreased at during apneic period compared to that at baseline (P<0.05). Conclusion : Through this experiment, we were partially able to understand the changes of cardiovascular function indirectly, but delete new experimental animal model displaying physiological mechanism close to natural sleep should be established, and the advanced study in the changes of cardiovascular function and their causes should be continued.

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