• Clinical and Experimental Pediatrics
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• v.50 no.10
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• pp.1011-1017
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• 2007

Purpose : We tried to assess the optimal conditions to improve low transduction efficiency and their effect on target cells. Methods : Cultured NIH 3T3 cells were incubated with retroviral vectors bearing an enhanced green fluorescent protein (eGFP) gene. We varied the ratio of viral vectors to target cells (1:1-1:8) and the number of transfections (${\times}1$, ${\times}2$), and compared transduction efficiencies. Also, the effects of polybrene on transduction efficiency and viability of target cells were assessed. Transduction of the eGFP gene was evaluated by observing NIH 3T3 cells under a fluorescence microscope and efficiencies were measured by the percentage of eGFP positive cells using FACscan. Results : As the ratio of retroviral vectors to target cells increased, transduction efficiency was greatly improved, from 7% (1:1) to 38% (1:4). However, transduction efficiency did not increase any more when the ratio increased from 1:4 to 1:8. Cells transfected twice showed higher transduction efficiencies than cells transfected once, at a ratio of 1:8. The eGFP gene transduced to NIH 3T3 cells sustained its expression during repeated passages. However, after the third passage (day 9), the percentage of eGFP positive cells began to decline. The degree of this decline in eGFP expression was lower in cells transfected twice than in cells transfected once (P<0.05). The addition of polybrene did not have any toxic effect on NIH 3T3 cells and greatly increased transduction efficiency (P=0.007). In addition to vector component, transduction efficiency was very sensitive to culture confluence. Cells cultured and transfected in 24-well plate showed higher transduction efficiency, although cells cultured in 6- well plate proliferated more (P=0.024). Conclusion : Our data could be used as a basis for retrovirus-based gene therapy. Further study will follow using human cells as target cells.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.32 no.6
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• pp.718-724
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• 1999

This study was investigated the changes of the properties of matter such as the gel formation of the combined fish based on the mixed rate between the ocean jumbo squid and Alaska pollock surimi, and compared the relationships between the gel formation and water holding capacity. The changes of the gel formation based on 20 min fish grinding time and $2.5\%$ salt concentration according to the mixed rate was thought as the optimal addition limit. There was no significant function of gel product more than $20\%$ Jumbo squid meat. The more squid meat in the mixed meat could make the lower breaking stress but 7:3 rate of pollock : squid could retain breaking strain. The effect of the moisture content on mixed fish meat was studied and the drastic decrease of the gel formation and water holding capacity was indicated in $78\%$.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.19 no.3
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• pp.219-226
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• 1986

To confirm the application of a newer in vitro assays to determining the nutritional value of marine crustaceans (mainly shrimps and crabs), which have been considered to be highly nutritive depending on their levels of the essential amino acids and digestibility, their C-PERs and DC-PERs were determined and studied the factors influencing their in vitro results. Four species of seawater shrimps and 2 species of seawater crabs were used in this experiment. The in vitro digestibilities showed $83{\sim}86\%$ for raw shrimps and the trypsin indigestibile substrate content (TIS) was ranged from 1.32 to 3.33 mg/g solid expressed quantitatively as mg of purified soybean trypsin inhibitor. The smaller size of shrimps revealed a greater in vitro digestibility and a lower contents of TIS. It was noted that the in vitro digestibility of raw blue crab meat was around $85\%$ while boiled tenner crab meat showed $86\%$ or above, and the leg meat had the greatest in vitro digestibility in the various parts of crab meats. The poor in vitro digestibilities for shrimp's and crab's meat, compared with that of the other seafoods as noted in previous reports, suggest that the drop in pH, due to the change in their freshness during harvesting and frozen storage, resulted in underestimating their digestibilities using four-enzyme digestion technique. The lysine contents in all samples were higher than that of ANRC casein but they contained a slightly lower sulfur-containing amino acids than those in ANRC casein. But the other EAA, such as valine, tyrosine and phenylalanine, were found to be a half as little as that in casein and played a key-factor in calculation of C-PER or DC-PER. It was observed that the value of C-PER and DC-PER for all samples ranged from 2.1 to 2.4, and the predicted digestibilities showed $90\%$ or above in all samples. It was a different results from the fact that the animal proteins bear a higher values and predicted digestibilities than those of C-PER values. The lack of correlation between C-PER and DC-PER values is attributable to the fact that the lower content of valine, tyrosine and phenylalanine, and drop in pH owing to the changes of freshness in marine crustacea proteins. Therefore, if a newer in vitro digestion technique-which are taken into account the pH drop before digestion, TIS content and released free amino acids and/or peptides-developed, C-PER assays can provide more advantages in assessing the protein nutritional value of marine crustacea than any other in vitro assays.

• Korean Journal of Animal Reproduction
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• v.24 no.4
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• pp.357-364
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• 2000

Members of the glycoprotein family, which includes CG, LH, FSH and TSH, comprise two noncovalently linked $\alpha$- and $\beta$-subunits. Equine chorionic gonadotropin (eCG), known as PMSG, has a number of interesting and unique characteristics since it appears to be a single molecule that possesses both LH- and FSH-like activities in other species than the horse. This dual activity of eCG in heterologous species is of fundamental interest to the study of the structure-function relationships of gonadotropins and their receptors. CG and LH $\beta$ genes are different in primates. In horse, however, a single gene encodes both eCG and eLH $\beta$ -subunits. The subunit mRNA levels seem to be independently regulated and their imbalance may account for differences in the quantities of $\alpha$ - and $\beta$-subunits in the placenta and pituitary. The dual activities of eCG could be separated by removal of the N-linked oligosaccharide on the $\alpha$-subunit Asn 56 or CTP-associated O-linked oligosaccharides. The tethered-eCG was efficiently secreted and showed similar LH-like activity to the dimeric eCG. Interestingly, the FSH-like activity of the tethered-eCG was increased markedly in comparison with the native and wild type eCG. These results also suggest that this molecular can implay particular models of FSH-like activity not LH-like activity in the eCG/indicate that the constructs of tethered molecule will be useful in the study of mutants that affect subunit association and/or secretion. A single-chain analog can also be constructed to include additional hormone-specific bioactive generating potentially efficacious compounds that have only FSH-like activity. The LH/CG receptor (LH/CGR), a membrane glycoprotein that is present on testicular Leydig cells and ovarian theca, granulosa, luteal, and interstitial cells, plays a pivotal role in the regulation of gonadal development and function in males as well as in nonpregnant and pregnant females. The LH/CGR is a member of the family of G protein-coupled receptors and its structure is predicted to of a large extracellular domain connected to a bundle of seven membrane-spanning a-helices. The LH/CGR phosphorylation can be induced with a phorbol ester, but not with a calcium ionophore. The truncated form of LHR also was down-regulated normally in response to hCG stimulation. In contrast, the cell lines expressing LHR-t631 or LHR-628, the two phosphorylation-negative receptor mutant, showed a delay in the early phase of hCG-induced desensitization, a complete loss of PMA-induced desensitization, and an increase in the rate of hCG-induced receptor down-regulation. These results clearly show that residues 632~653 in the C-terminal tail of the LHR are involved in PMA-induced desensitization, hCG-induced desensitization, and hCG-induced down-regulation. Recently, constitutively activating mutations of the receptor have been identified that are associated with familial male-precocious puberty. Cells expressing LHR-D556Y bind hCG with normal affinity, exhibit a 25-fold increase in basal cAMP and respond to hCG with a normal increase in cAMP accumulation. This mutation enhances the internalization of the free and agoinst-occupied receptors ~2- and ~17- fold, respectively. We conclude that the state of activation of the LHR can modulate its basal and/or agonist-stimulated internalization. Since the internalization of hCG is involved in the termination of hCG actions, we suggest that the lack of responsiveness detected in cells expressing LHR-L435R is due to the fast rate of internalization of the bound hCG. This statement is supported by the finding that hCG responsiveness is restored when the cells are lysed and signal transduction is measured in a subcellular fraction (membranes) that cannot internalize the bound hormone.

• Journal of Animal Science and Technology
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• v.44 no.6
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• pp.757-768
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• 2002

These study were conducted to determine the effects of a whole or steam-flaked corn based diet on rumen microbial fermentation in vitro and ruminal metabolism in the Korean Native Goat(KNG) in vivo. The experiments consisted of two dietary treatments: control, steam-flaked corn(SFC) based diet(80%) + rice straw mixed(20%)(SFCR); 100% whole corn based diet(WC). The first experiment was conducted to investigate the effect of whole corn on ruminal metabolism in vitro for 0 to 48 h. pH values were optimally maintained during incubation time, and were not significantly different between treatments. Gas production of SFCR was significantly higher than WC(p<0.01). $NH_3$-N concentration tended to increase for WC, but not significantly different between treatments. The mean value of total volatile fatty acid concentration of WC was significantly lower than SFCR(p<0.01), but SFCR and WC linearly increased as the time of incubation approached 48 h. Mean value of acetate concentration of SFCR was significantly higher than WC(p<0.01). Propionate concentration of WC for the total incubation time was significantly higher than SFCR(p<0.01). The digestibility of dry matter was not significantly different between treatments, but SFCR was somewhat higher than WC. The second experiment was conducted to effect of whole shelled corn based diet on rumen metabolism in KNG. pH values tended to decrease through all treatments. There was not a significantly difference between treatments. Microbial protein yield of SFCR was significantly higher than WC(p<0.01). $NH_3$-N concentration of WC was significantly (p<0.01) higher than SFCR. Total VFA and propionate concentration of WC was significantly higher than SFCR(p<0.01), but acetate concentrate of WC was not significantly higher than SFCR. The mean value of total lactate concentration was significantly(p<0.01) different but the value of SFCR and WC were lower than the average concentration of acidosis. In sacco DM disappearance rate of SFC was significantly(p<0.01) higher than WC.

• Journal of Animal Science and Technology
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• v.47 no.4
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• pp.565-572
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• 2005

An experiment was conducted to determine the effect of supplementation of insoluble dietary fiber (Vitacel®) on growth performance and nutrient digestibility in weanling pigs. A total of 96 pigs that averaged 6.49$\pm$0.52kg BW and 23$\pm$2.1d age were allocated in a randomized block design with two pigs per pen and 12 pens per treatment. Pigs and feeders were weighed 10-days interval for the 40-d trials to determine ADG, ADFI and feed:gain ratio(F:G). Pigs were fed one of four diets:1) Control diet (C) 2) C+0.3% insoluble dietary fiber(IDF) 3) C+0.6% IDF and 4) c+0.9% IDF. For the determination of fecal nutrients digestibility, pigs were fed diets(diet 2) with 1% Celite-545(Fluka) as a marker and feces were collected on $9^{th}$ day and $18^{th}$ day after feeding diet 2. During the whole experimental period, pigs fed diet with 0.3% IDF have significantly higher ADG than other dietary treatment groups(P<0.05). ADG of pigs fed diet with 0.6% IDF was higher than that of pigs fed control diet(P<0.05). However, there was no significant difference in ADG between control group and 0.9% IDF group(P>0.05). ADFI of pigs fed diet with 0.3% IDF was significantly higher than any other dietary treatment groups(P<0.05). There was no significant difference in ADFI among control group, 0.6% and 0.9% IDF supplementation groups (P>0.05). Digestibilities of organic matter, crude protein, crude fiber and energy were significantly higher in 0.3% IDF supplementation group than any other dietary groups. However, there was no difference in over all nutrient digestibilities between 0.6% and 0.9% IDF group. Feeding diets more than 0.6% IDF did not affect the rest of the nutrients digestibilities except for ADF digestibility compared to control diet. Dietary supplementation level of IDF showed a significant quadratic effect on performance improvement of piglets. This response of growth performance to IDF supplementation is, as expected, in agreement with that of nutrient digestibility. Our results showed that IDF supplementation to diet for weaned piglets might be beneficial in terms of growth and nutrient digestibility. However, there should be more study on the relationship between level of IDF supplementation and piglet response as well as the exact mode of action of IDF in weaned piglets.

• Journal of Animal Science and Technology
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• v.45 no.3
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• pp.387-396
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• 2003

This experiment was carried out to compare the effects of feeding commercial formula feed and rice straw separately (control) versus a total mixed ration (TMR) on productivity of Hanwoo steers in late stage of fattening and on ruminal fermentation characteristics and digestibilities. Ruminal digesta from the cannulated cattle were sampled at 0, 1, 2, 3, 5, 8 hour after feeding. The steers fed TMR consumed 7.4kg per day and there was no difference between feeding systems. Daily weight gain was not significantly (P>0.05) between feeding systems, however, TMR group showed lower daily gain than control group. The amount of feed consumption per kg weight gain was higher in TMR group than control group (10.5kg and 9.7kg, respectively), resulting in a greater efficiency of feed utilization for gain. In the result of appearance rates of quality grade A were 33% higher for TMR group than those in control group. Appearance rates of grade 1 showed 56% and 75% when fed the control and TMR, respectively. Digestibilities of dry matter, crude protein, crude fiber and gross energy for TMR treatment were significantly higher (P<0.01) than those of control. Prior to feeding (0 h) and each subsequent hour, the TMR resulted in higher rumen pH (P<0.05) when compared with control ration. The concentration of NH3-N for TMR treatment maintained at higher level up to 8hr after feeding, especially increased up to 28.2mg/$d\ell$ during 1-2 hour which was two times (P<0.05) more than control. The amount of total VFA showed same trends between feeding systems. However, the ratios of branched chained fatty acid such as iso-butyric acid and iso-valeric acid for TMR treatment were significantly(P<0.01) higher than control for 3-5hr. Results showed that TMR in these trials is effective feeding system for fattening Hanwoo steers in the respect of ruminal characteristics, total tract digestibility and productivities.

• Clinical and Experimental Pediatrics
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• v.50 no.3
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• pp.284-291
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• 2007

Purpose : Theophylline has recently been reported to have concurrent anti-inflammatory effects at low therapeutic plasma concentrations which are below the doses at which significants, clinically useful bronchodilatation is evident. Sustained-release formulation in capsule and dry syrup forms were developed to reduce its adverse effects and improve its clinical effects. We compared the therapeutic effects of theophylline dry syrup and capsules in children with mild asthma. Methods : Ninety children with mild asthma were randomized to receive either theophylline dry syrup (n=44) or theophylline capsules (n=46); 4 mg per kilogram of body weight, twice a day, for 12 weeks. Baseline and serial measurements of daytime and nighttime asthma symptom score were performed. Compliance scores, drug swallowing scores, and drug usability scores were measured every 4 weeks. Each scoring was rated on a scale of 0-4. Serum theophylline concentration were measured at 4 and at 12 weeks. To examine the anti-inflammatory effect of theophylline on asthma, Serum eosinophilic cationic protein as a marker of airway inflammation caused by eosinophil was measured 12 weeks pre- and post-administration. Results : The daytime and nighttime asthma symptom scores of the two groups after 4 weeks significantly improved over the baseline score. Daytime and nighttime asthma symptom scores in the dry syrup group were statistically lower at all time points except for the nighttime symptom scores at 4 weeks. Compliance scores, drug swallowing scores, and drug usability scores in the dry syrup group were significantly higher at the end time point. Only in the dry syrup group was the serum ECP at the end time point statistically lower than baseline. Conclusion : Low-dose sustained-release theophylline may be safe and effective in bronchial asthma and this effect may be mediated by its anti-inflammatory action mechanisms. Especially, when used in children with asthma, dry syrup formulation is recommended because of its higher compliance than capsule formulation.

• Journal of the Korea Organic Resources Recycling Association
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• v.9 no.1
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• pp.65-72
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• 2001

Four newly isolated bacteria from soil were used to manufacture microbial inoculum to compost food waste. The bacteria, GM103, V25, V31, and V35, were identified as Bacillus licheniformis, B. subtilis, B. stearothermophilius, and B, subtilis, respectively. The bacterial strains were efficient to degrade protein and starch and also able to inhibit the growth of plant pathogenic fungus Rhizopus stronifer. The GM103 showed distinct capability in degrading starch, but grow only aerobically. The other three bacterial strains. V25, V31, and V35, could grow both aerobically as well as anaerobically, in 10%(w/v) salt, at $50^{\circ}C$, and had good viability and survival rate in soil. These characteristics of the bacterial strains are very adquate in Korean food composting containing high concentration of salt, especially at home. By mixing the 4 bacterial culture broth with molasses, beet pulp, zeolite, The bacterial inoculum for food waste composting-BIOTOP-CLEAN-was made. The performance of food waste composting by the BIOTOP-CLEAN was compared with that by control(not treated) and HS(other demestic company's inoculum product for food waste composting). The maximum temperature of the food waste during the composting with the BIOTOP-CLEAN was $50^{\circ}C$, while those of the control and HS were $30^{\circ}C$ and $35^{\circ}C$, respectively. The BIOTOP-CLEAN gave the good smell and showed dark brown color, while the control gave bad smell and HS gave less bad smell. These indicates that the food waste composting by the BIOTOP-CLEAN had been well accomplished. The culture broth of V25, V31, V35 were sparyed to the plants of tomato, chinese cabbage, raddish, red pepper every month and the spraying the culture broth to these plant significantly improved the production yield of the crops, due to the control effect of the bacterial strains against the plant pathogens.

• Proceedings of the Korean Society for Applied Microbiology Conference
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• 2005.06a
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• pp.154-164
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• 2005

Saccharomyces cerevisiae KNU5377 is a thermotolerant strain, which can ferment ethanol from wasted papers and starch at 40$^{\circ}C$ with the almost same rate as at 30$^{\circ}C$. This strain showed alcohol fermentation ability to convert wasted papers 200 g (w/v) to ethanol 8.4% (v/v) at 40$^{\circ}C$, meaning that 8.4% ethanol is acceptable enough to ferment in the industrial economy. As well, all kinds of starch that are using in the industry were converted into ethanol at 40$^{\circ}C$ with the almost same rate as at 30$^{\circ}C$. Hyperthermic cell killing kinetics and differential scanning calorimetry (DSC) revealed that exponentially growing cells of this yeast strain KNU5377 were more thermotolerant than those of S. cerevisiae ATCC24858 used as a control. This intrinsic thermotolernace did not result from the stability of entire cellular components but possibly from that of a particular target. Heat shock induced similar results in whole cell DSC profiles of both strains and the accumulation of trehalose in the cells of both strains, but the trehalose contents in the strain KNU5377 were 2.6 fold higher than that in the control strain. On the contrary to the trehalose level, the neutral trehalase activity in the KNU5377 cells was not changed after the heat shock. This result made a conclusion that though the trehalose may stabilize cellular components, the surplus of trehalose in KNU5377 strain was not essential for stabilization of whole cellular components. A constitutively thermotolerant yeast, S. cerevisiae KNU5377, was compared with a relatively thermosensitive control, S. cerevisiae ATCC24858, by assaying the fluidity and proton ATPase on the plasma membrane. Anisotropic values (r) of both strains were slightly increased by elevating the incubation temperatures from 25$^{\circ}C$ to 37$^{\circ}C$ when they were aerobically cultured for 12 hours in the YPD media, implying the membrane fluidity was decreased. While the temperature was elevated up to 40$^{\circ}C$, the fluidity was not changed in the KNU5377 cell, but rather increased in the control. This result implies that the plasma membrane of the KNU5377 cell can be characterized into the more stabilized state than control. Besides, heat shock decreased the fluidity in the control strain, but not in the KNU5377 strain. This means also there's a stabilization of the plasma membrane in the KNU5377 cell. Furthermore, the proton ATPase assay indicated the KNU5377 cell kept a relatively more stabilized glucose metabolism at high temperature than the control cell. Therefore, the results were concluded that the stabilization of plasma membrane and growth at high temperature for the KNU5377 cell. Genome wide transcription analysis showed that the heat shock responses were very complex and combinatory in the KNU5377 cell. Induced by the heat shock, a number of genes were related with the ubiquitin mediated proteolysis, metallothionein (prevent ROS production from copper), hsp27 (88-fold induced remarkably, preventing the protein aggregation and denaturation), oxidative stress response (to remove the hydrogen peroxide), and etc.