The main goal of this study was to produce transgenic piglets by the method of injection of sperm-mediated exogenous DNA. Spermatozoa (1$\times$106 sperm of final concentration) obtained from caudal epididymis were mixed with pBC1-hEPO (20 ng/${mu}ell$) or pcDNA3 LAC Z (20 ng/${mu}ell$), and followed by electroporation (500 V, 25 ㎌). Matured oocytes having the first polar body and dense cytoplasm were selected and centrifuged at 12,000g for 6 min. After sperm injection, the oocytes were activated electrically (1.7 ㎸/cm, 30 $\mu$ sec, single pulse) in 0.3 M mannitol solution. Eggs injected sperm were cultured in NCSU 23 medium (0.4% BSA) at 39$^{\circ}C$, 5% $CO_2$ in air for 192 h. This study were comprised 3 experiments. Experiment 1 compared the developmental efficiencies between the sperm-injected oocytes (Group 1) and further activated electrically (Group 2). Experiment 2 compared the expression of pcDNA3 LAC Z in the embryos produced by Group 1 and Group 2. Finally, experiment 3 carried out transfer of embryos (1-8 cell stage) transfected with pBC1 -hEPO into surrogate recipients synchronized by injection of combination of PG600 with hCG. The rates of cleavage and development into blastocyst stage in Group 2 were significantly higher than those of Group 1 (71.3% and 28.1% vs. 43.3% and 10.3%, respectively, p<0.05). Thirty (24.2%) out of 124 embryos analyzed in Group 2 were positive by X-gal. Similarly, in Group 1, 16.3% (8/49) were positive. After transfer of 789 embryos to 7 recipient gilts, three out of them examined by ultrasound became pregnant. One recipient is in day 50 pregnancy. On day 54 of gestation, two were carried out uterotomy in order to confirm the pregnancy One had 7 and another had 2 fetuses. We conclude that injection of sperm-mediated gene transfer will be used as a valuable tool for the production of transgenic piglets.
In the present study the effect of exercise on the conversion rate of ingested glucose to glycogen in the different types of hindlimb skeletal muscles in Sprague-Dawley male rats was studied. The amounts of synthetized glycogen from ingested glucose of fast-twitch white(WV), fast-twitch red(RV), mixed type of fast-twitch white and red(EDL), and slow-twitch(SOL) muscles were determined at 30 and 90 min. after ingestion of 25% glucose solution which contained $^{14}C$-glucose($2m{\ell}(1uCi)$/100gm B. W.)in control and exercise loaded rats. The result was summarized as follows : The about 55% at 30 min. and 70% at 90 min. after glucose ingestion were absorbed from gastrointestinal tract. This result shows no effects of exercise on absorption rate from gastrointestinal tract. The amount of synthetized glycogen of SOL from ingested glucose at 30 and 90 min. after glucose ingestion were highest, whether WV were lowest in hindlimb skeletal muscles in control and exercise loaded rats. In the exercise loaded rats, the amounts of synthetized glycogen of SOL, RV, and EDL at 90 min. after glucose ingestion was much higher than control rats, but not different in WV between exercise-loaded and control rats. At 30 min. after glucose ingestion, only SOL of exercise loaded rats was higher than control rats. In the control rat, the synthesis of glycogen was almost completed during initial 30 minutes. On the other hand, in the exercise loaded rat, except WV was opposite result of control rats, i. e., amounts of synthetized glycogen were major during late period. The amount of synthetized glycogen of liver at 30 and 90 min. after glucose ingestion in exercise loaded rats was higher than control rats. The rate of glycogen synthesis in control and exercise loaded rats were higher between 30-90 minute than initial 30 minute.
Sunagawa, K.;Ooshiro, T.;Nakamura, N.;Nagamine, I.;Shiroma, S.;Shinjo, A.
Asian-Australasian Journal of Animal Sciences
/
v.18
no.10
/
pp.1414-1420
/
2005
The purpose of this research was to determine whether or not feeding induced hypovolemia (decreases in plasma volume) and decreases in plasma bicarbonate concentration caused by loss of $NaHCO_3$ from the blood, act to suppress feed intake and saliva secretion volumes during the initial stages of feeding in goats fed on dry forage. The animals were fed twice a day at 10:30 and at 16:00 for 2 h each time. Prior to the morning feeding, the collected saliva (3-5 kg) was infused into the rumen. During the morning 2 h feeding period (10:30 to 12:30), the animals were fed 2-3 kg of roughly crushed alfalfa hay cubes. At 16:00, the animals were fed again with 0.8 kg of alfalfa hay cubes, 200 g of commercial ground concentrate and 20 g of sodium bicarbonate. In order to compensate for water or $NaHCO_3$ lost through saliva during initial stages of feeding, a 3 h intravenous infusion (17-19 ml/min) of artificial mixed saliva (ASI) or mannitol solution (MI) was begun 1 h prior to the morning feeding and continued until the conclusion of the 2 h feeding period. The physiological state of the goats in the present experiment remained unchanged after parotid gland fistulation. Circulating plasma volume decreases caused by feeding (estimated by increases in plasma total protein concentration) were significantly suppressed by the ASI and MI treatments. During the first 1 h of the 2 h feeding period, plasma osmolality in the ASI treatment was the same as the NI (non-infusion control) treatment, while plasma osmolality in the MI treatment was significantly higher. In comparison to the NI treatment, cumulative feed intake levels for the duration of the 2 h feeding period in the ASI and MI treatments increased markedly by 56.6 and 88.3%, respectively. On the other hand, unilateral cumulative parotid saliva secretion volume following the termination of the 2 h feeding period in the ASI treatment was 50.7% higher than that in the NI treatment. MI treatment showed the same level as the NI treatment. The results of the present experiment proved that the humoral factors involved in the suppression of feeding and saliva secretion during the initial stages of feeding in goats fed on dry forage, are feeding induced hypovolemia and decrease in plasma $HCO_3^-$ concentration caused by loss of $NaHCO_3$ from the blood.
Journal of Korean Society of Environmental Engineers
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v.22
no.3
/
pp.523-533
/
2000
A two-phase anaerobic reactor with submerged membrane system was developed for increasing acidogen concentration and methane recovery. The membrane used was mixed esters of cellulose of $0.5{\mu}m$ pore size and $0.8m^2$ of effective surface area. The methanogenic reactor comprised of UASB (Upflow Anaerobic Sludge Blanket) and AF (Anaerobic Filter). COD removal efficiency was 70~80% and the methane content in the biogas increased up to 90% for the submerged membrane system in the anaerobic reactor. As the cake resistance of membrane caused a serious problem, stainless steal prefilters (40, 53, $63{\mu}m$) and air backwashing methods were applied to minimize the cake resistance effectively. Among the tested prefilters. the $63{\mu}m$ prefilter showed the best performance for reduction of cake resistance and a successful long-tern operation. By cleaning with alkali first and acidic solution later. the permeate flux decreased by long term operation was recovered to 89% of that with a new membrane.
Journal of Korean Society for Atmospheric Environment
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v.29
no.1
/
pp.97-104
/
2013
Methyl alcohol is one of the basic intermediates in the chemical industry and is also being used as a fuel additive and as a clean burning fuel. In this study, conversion of carbon dioxide to methyl alcohol was investigated using catalytic chemical methods. Ceramic monoliths (M) with $400cell/in^2$ were used as catalyst supports. Monolith-supported CuO-ZnO catalysts were prepared by wash-coat method. The prepared catalysts were characterized by using ICP analysis, TEM images and XRD patterns. The catalytic activity for carbon dioxide hydrogenation to methyl alcohol was investigated using a flow-type reactor under various reaction temperature, pressure and contact time. In the preparation of monolith-supported CuO-ZnO catalysts by wash-coat method, proper concentration of precursors solution was 25.7% (w/v). The mixed crystal of CuO and ZnO was well supported on monolith. And it was known that more CuO component may be supported than ZnO component. Conversion of carbon dioxide was increased with increasing reaction temperature, but methyl alcohol selectivity was decreased. Optimum reaction temperature was about $250^{\circ}C$ under 20 atm because of the reverse water gas shift reaction. Maximum yield of methyl alcohol over CuO-ZnO/M catalyst was 5.1 mol% at $250^{\circ}C$ and 20 atm.
The purpose of this study is to examine trends and achievements of domestic gamification studies and to suggest further research directions. For this purpose, I collect references in domestic gamification and then analyze them through a systematic literature review. Through search DB, 131 references were collected, the references were analyzed by year, purpose, research type, research methods, and research results. The results showed that the amount of gamification research has increased steadily, and academic discussion in the non-game context has increased sharply since 2015. The results of analysis by research purpose showed that the most research was conducted to evaluate the effectiveness of the strategy, program, and design of gamification. The results of research type showed solution, evaluation, position paper, and validation study in that order. In terms of research methods, literature research was the most common, followed by quantitative, qualitative and mixed research. In the analysis of 45 papers that reported the research results, it was found that there were 21 articles about psychological effects, 17 articles about behavioral effects, and 7 articles about psychological and behavioral effects. Further research directions are suggested based on the review results.
This study was conducted to develop an intrauterine inseminator (IUI) to deposit of frozen semen into uterus and to evaluate the results obtained after artificial insemination by IUI. Two Japanese spitzs (2 to 4 years of age) were used as semen donors. Semen was collected by manual masturbation into sterile glass collection tubes and separated into 3 fractions with only the sperm-rich fractions retained for further examination. Sperm motility >70%, sperm concentration of 200 to $400{\times}10^6 cells/ml$$\times$g for 5 min and poured out the suspended solution, and then diluted with 2 ml Tris-buffer which was consisted of 2.4 g Tris, 1.4 g citric acid, 0.8 g glucose, 0.1 $\mu\textrm{g}$/ml streptomycin, 100 IU/ml penicillin, 20 ml egg yolk to 100 ml mili-Q water (Ext I) or supplemented with 8 ml glycerol and 1 ml Equex STM paste to 100 rnl (Ext II). The diluted semen was cooled to 5$^{\circ}C$ in cold room, where the temperature in the sample reached 5$^{\circ}C$. Two h after beginning the cooling procedure, 2 ml of Ext II, also at 5$^{\circ}C$, was added and mixed by gently reversing the tubes several times during 1 h. The final sperm concentration for freezing was approximately $50{\times}10^6 cells/ml$. After equilibration, the semen was loaded into 0.5 ml straw and frozen on the liquid nitrogen vapour in styrofoam box. The straws were thawed at 7$0^{\circ}C$ for precisely 6 sec. After thawing of each straw, the frozen semen can survived over 50% motility. All the females were inseminated twice with 1 ml of $25{\times}10^6 cells/ml$
Park, Cheol-Ho;Yu, Chang-Yeon;Seo, Jeong-Sik;Kim, Ki-Sik;Park, Seung-Ue;Chang, Byoung-Ho;Ahn, Sang-Deuk
Korean Journal of Medicinal Crop Science
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v.3
no.3
/
pp.207-216
/
1995
This study was conducted to establish technologies for plug seedling production using somatic embryos-derived regenerants and their seeds in Bupleurum falcatum L. Among distilled water, GA (0.1mg/l) and putrescine (0.lmg/l) treated to regenerants for acclimatization, GA was most effective to develop shoots and roots, 1/2X MS medium and NAA 0.1mg/l + BA 0. 5mg/l enhanced the growth rates of the regenerants and increased dry weight. Activated charcoal effected to grow markedly leaves and roots of the regenerants at the level of 0.4 %. Regenerants increased their plant height, root length and dry weight at $30^{\circ}C$. Plug seedlings originated from seeds of the tissue culture regenerants showed the maxium growth on the mixture of peatmoss soil (2) and mountain sand (1) .Root length, leaf area and dry weight of plug seedlings increased significantly when No.1, 2 and 3 of Wondergrow solution were mixed in the ratio of 1.3 - 0.9 - 0.1. Light supplement (4%) and high temperature $(30^{\circ}C)$ promoted the growth of plug seedlings as well as dry weight. Ninety days seedlings were more vigorous and adaptable for transplanting than other seedlings.
Objectives : The purpose of this study is to evaluate effect of suppressing the expression of cyclo-oxygenase-type-2 (COX-2) as a consequence of inhibition macrophage migration inhibitory factor (MIF) activation by $Sambucus$$williamsii$$Hance$ (SWH) pharmacopunctureon rheumatoid arthritis (RA). Methods : In vitro test, synoviocytes extracted from type II collagen-induced arthritis (CIA) mouse's knee joint were cultivated After that, each well of synoviocytes was mixed with the extract of SWH at the dosage of $0.4mg/m{\ell}$, $0.6mg/m{\ell}$, $0.8mg/m{\ell}$, and $1.0mg/m{\ell}$ respectively, and cultivated for 24 hours after the addition. Reverse transcriptase - polymerase chain reaction (RT-PCR) is used to investigate the expression of MIF, Tumor necrosis factor (TNF)-${\alpha}$, COX-2 mRNA. $In$$vivo$ test, thirty DBA female mice were used, and each ten mice were allocated into three group; normal group, CIA-elicitated group, and group treated with SWH pharmacopuncture on it's the point of $ST_{35}$ after CIA elicitation. It is investigated that change of mice foot thickness, histologic change of sliced synovial joint of mouse, and extent change of MIF, TNF-${\alpha}$, COX-2 in synovial membrane. Results : $In$$vitro$ test, the expressions of cytokine(MIF, TNF-${\alpha}$, COX-2) mRNAs related to RA were dose-dependent decreased. In the SWH pharmacopuncture group, foot thickness and histologic change of sliced synovial joint were decreased comparing with CIA-elicitated group's change. In the SWH pharmacopuncture group, the suppression of MIF, TNF-${\alpha}$, COX-2 in synovial membrane was clearly shown comparing with CIA-elicitated group's change. Conclusions : It might be suggested that SWH pharmacopuncture mitigate tissue damage originated from rheumatoid arthritis by suppressing the expression of COX-2 as a consequence of inhibition MIF activation.
Journal of the Korean Crystal Growth and Crystal Technology
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v.23
no.5
/
pp.241-245
/
2013
Granular bamboo-derived active carbons (AC) were impregnated (or coated) with $TiO_2$ nano crystalline powders. The photocatalytic activity of the $TiO_2$-impregnated active carbons ($TiO_2$/AC) were determined on the basis of the degradation rate of methylene-blue aqueous solution under UV irradiation. The active compounds of $TiO_2$ were impregnated onto the AC under moderate hydrothermal conditions (${\leq}200^{\circ}C$, pH 11). The mean size of $TiO_2$ particles calculated from BET surface area were found to be as 50 nm. The $TiO_2$ precipitates were coated on the cavities or pores on the surfaces of highly activated carbons. Since the hydrothermal process led to a lowering of the on-set temperature of the anatase-to-rutile transition of $TiO_2$ as low as $200^{\circ}C$, $TiO_2$ crystallites of a pure anatase or a mixed form with rutile were successfully coated on the AC depending on the synthesis temperatures.
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