• KSBB Journal
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• v.31 no.4
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• pp.284-290
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• 2016

In this research, recombinant human cytotoxic T-lymphocyte antigen 4-immunoglobulin (hCTLA4Ig) was produced by transgenic rice cells. RAmy3D promoter was used for overcome the limitation of low expression level in transgenic plant cells, and the secretion of target protein was accomplished by signal peptide. However, the RAmy3D promoter system which can be induced only by sugar starvation causes the decrease of cell viability. As a result, cell death promotes the release of protease which degrades the target proteins. The protein stability and productivity can be significantly influenced by proteolysis activity. Therefore, development of new strategies are necessary for the in situ recovery of target proteins from cell culture media. In this study, in situ recovery was performed by various strategies. Direct addition of Protein A resin with nylon bag leads to cell death by increased shear stress and decrease in production of hCTLA4Ig by protease. Medium exchange through modified flask could recover hCTLA4Ig with high cell viability and low protease activity, on the other hand, the productivity was lower than that of control. When in situ recovery was conducted at day 7 after induction in air-lift bioreactor, 1.94-fold of hCTLA4Ig could be recovered compared to control culture without in situ recovery. Consequently, in situ recovery of hCTLA4Ig from transgenic rice cell culture could enhance productivity significantly and prevent degradation of target proteins effectively.

• Journal of Life Science
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• v.12 no.5
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• pp.535-543
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• 2002

To investigate the possibility of processing semi-dried jellyfish and to examine its keeping quality, the factors such as moisture content, pH, volatile basic nitrogen (VBN), viable cell counts, color value and texture were analyzed. For processing of the semi-dried product, optimum conditions of drying temperature and drying time were $25^{\circ}C$ and 90min, respectively, under 60 $\pm$ 5% of relative humidity and 2 m/sec of air blast speed. The shelf-life of the products, vacuum-packing method in nylon/polyethylene/linear low density polyethylene (0.015/0.045/0.040 mm) laminated film bag was at least 6 weeks during storage at 4$^{\circ}C$, while that of air packaged one was at least 4 weeks. In case of 2$0^{\circ}C$ storage the shelf-life of the products was at least 3 weeks in both air and vacuum packaged ones.

• Asian-Australasian Journal of Animal Sciences
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• v.13 no.3
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• pp.313-316
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• 2000

Three Holstein steers, fitted with ruminal and duodenal cannulas, were used in a replicated $3{\times}3$ Latin square design to determine the digestibility of dietary nitrogen in total digestive tract by three methods, 1) mobile nylon bag (MNB); 2) total fecal collection (TFC); and 3) washed fecal sample after freezing and thawing through a sieve with a pore size of $45{\mu}m$ (WFS). A basal diet of oaten hay-barley was supplemented with one of the following protein sources; soybean meal, fish meal or blood meal. Steers were fed at a level of 2% of body weight. The experimental diets were contained approximately 1.85% nitrogen. There were no differences (p>0.05) among the diets on DM, NDF and nitrogen disappearances, and the diet results were pooled to assess the methods. Total tract disappearances of dry matter and neutral detergent fiber were 61.6, 71.1 and 78.9 and 25.3, 63.2 and 64.6 for MNB, TFC and WFS methods, respectively. The lower digestibility of DM and NDF in the MNB method could be a result of low ruminal incubation time. The TFC method had the lower (p<0.05) determination of nitrogen disappearance in the total digestive tract than the MNB and WFS methods. On the other hand, nitrogen disappearance in the total digestive tract determined by the WFS technique was comparable to that in MNB technique, as there was no significant difference (p>0.05) between the methods. It is shown that the disappearance of dietary nitrogen in the total digestive tract could be estimated in the intact animals by using washed fecal sample prior to freezing and thawing.

• Asian-Australasian Journal of Animal Sciences
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• v.7 no.3
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• pp.389-396
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• 1994

Effects of chemical treatments on in sacco and in vitro digestibility of barley straw by rumen fluid and pure cultures of cellulolytic bacteria were studied to evaluate the pretreatment and to improve the poor quality feed. Chemicals were applied by dissolving them in water equivalent to 40% of the weight of the straw (dry matter basis). Pretreatment with 5% NaOH yielded the largest increase in sacco digestion followed by pretreatment with 2% $(NH_4)_2SO_3$, 2.6% $NH_4OH$, 1.6% $NaHSO_3$ and untreated straw (control). In sacco dry matter digestibility of straw treated with NaOH and $(NH_4)_2SO_3$ continued to increase as the concentration of chemical increased (1 to 7.5%), as it was the in vitro dry matter loss by leaching. Treatment of barley straw with 5% NaOH enhanced significantly (p < 0.01) in vitro digestibility by rumen fluid, Fibrobacter suceinogenes and Ruminococcus albus though the fermentation products by cellulolytic bacteria were low, whereas the treatment with 5% $(NH_4)_2SO_3$ inhibited in vitro digestibility by F. succinogenes and R. albus together with lower fermentation products. Dry matter loss by leaching and bacterial digestion from barley straw treated with NaOH and $(NH_4)_2SO_3$ suggested the effect of pretreatment with these chemicals were based on leaching, and the cellulolytic bacteria had little to do with digestion.

• Asian-Australasian Journal of Animal Sciences
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• v.19 no.2
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• pp.181-188
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• 2006

The objective of this research was to determine the effects of synchronizing the rate of dietary energy and nitrogen release on: ruminal fermentation, microbial protein synthesis, blood urea nitrogen, and nutrient digestibility in beef cattle. Four, two-and-a-half year old Brahman-Thai native crossbred steers were selected for the project. Each steer was fitted with a rumen cannula and proximal duodenal cannula. The steers were then randomly assigned in a $4{\times}4$ Latin square design to receive four dietary treatments. Prior to formulation of the dietary treatments, feed ingredients were analyzed for chemical composition and a nylon bag technique was used to analyze the treatments various ingredients for degradability. The treatments were organized in four levels of a synchrony index (0.39, 0.50, 0.62 and 0.74). The results showed that dry matter digestibility trend to be increased (p<0.06), organic matter and acid detergent fiber digestibility increased linearly (p<0.05), while crude protein and neutral detergent fiber digestibility were not significantly different (p>0.05). Higher concentration and fluctuation of ruminal ammonia and blood urea were observed in the animal that received the lower synchrony index diets. As the levels of the synchrony index increased, the concentrations of ruminal ammonia nitrogen and blood urea nitrogen, at the 4 h post feeding, decreased linearly (p<0.05). Total volatile fatty acid and bacteria populations at the 4 h post feeding increased linearly (p<0.05). Microbial protein synthesis trend to be increase (p<0.08). The results of this research indicate that synchronizing the rate of degradation of dietary energy and nitrogen release improves ruminal fermentation, microbial protein synthesis and feed utilization.

• Asian-Australasian Journal of Animal Sciences
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• v.19 no.8
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• pp.1120-1126
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• 2006

Dry matter (DM) degradation of leaves from Quercus cercis, Quercus libari, Quercus branti, and Quercus coccifera was determined using two different techniques: (i) in vitro gas production and (ii) the nylon bag degradability technique. In vitro gas production in the presence or absence of PEG and in situ DM disappearance were measured at 3, 6, 12, 24, 48, 72 and 96 h. In situ and in vitro DM degradation kinetics were described using the equation y = a+b ($1-e^{-ct}$). At all incubation times leaves from Quercus branti incubated with or without PEG gave significantly higher gas production than the other oak leaves except for 3 and 6 h incubation when leaves from Quercus branti without PEG supplementation only gave higher gas production than Quercus cercis and Quercus coccifera. At all incubation times except at 3, 6 and 12 h the DM disappearance from Quercus branti was significantly higher than the other species. Generally, PEG supplementation considerably increased the gas production at all incubation times and estimated parameters such as gas production rate ($c_{gas}$), gas production (ml) from the quickly soluble fraction ($a_{gas}$), gas production (b) from the insoluble fraction, potential gas production (a+b). However, all oak leaves did not give the same response to the PEG supplementation. Although the increase in gas production at 96 h incubation time was 8.9 ml for Quercus libari the increase was 5.5 ml for Quercus coccifera. It was concluded that except at early incubation times the relationships between the two methodologies seem to be sufficiently strong to predict degradability parameters from gas production parameters obtained in the presence or absence of PEG.

• Journal of Life Science
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• v.9 no.1
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• pp.44-49
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• 1999

A study was conducted to investigate the possibility of processing seasoned and semi-dried right-eyed flounder (Pseudopleuronects herzensteini) and to examine its quality during storage at $\pm$$3^{\circ}C$ by analyzing the factors such as moisture content, pH, volatile basic nitrogen (VBN), viable cell count and peroxide value (POV). For processing of the seasoned and semi-dried product, optimum conditions of drying temperature and drying time were $20^{\circ}C$ and 4 hrs, respectively, under 50$\pm$5$\%$ of relative humidity and 2 m/sec of air blast speed. The shelf-lives of the products by vacuum-packaging method in nylon/polyethylene/linear low density polyethylene (0.015/0.045/0.040 mm) laminated film bag were 35 and 16 days during storage at $-3^{\circ}C$ and $3^{\circ}C$, respectively.

• Korean Journal of Food Science and Technology
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• v.29 no.5
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• pp.925-931
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• 1997

An attempt was made in this study to investigate the possibility of processing seasoned and semi-dried horse mackerel (Tracharus japonnicus) and to examine its keeping quality, by analyzing the factors such as water content, pH, volatile basic nitrogen, viable cell count, peroxide value, color value and sensory evaluation. For processing of the seasoned and semi-dried product, optimum conditions of drying temperature and drying time were $20^{\circ}C$ and 4 hr, respectively, under $50{\pm}5%$ of relative humidity and 2 m/sec of air blast speed. The shelf-life of the products, vacuum-packaging method in nylon/polyethylene/linear low density polyethylene $(0.015/0.045/0.04\;{\mu}m)$ film bag, was 42 days during storage at $-3{\pm}0.5^{\circ}C$, while that of unpackaged one was only 28 days.

• Asian-Australasian Journal of Animal Sciences
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• v.22 no.1
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• pp.82-89
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• 2009

Two in vitro experiments were conducted to examine the effects of propionate precursors in the dicarboxylic acid pathway on ruminal fermentatation characteristics, $CH_4$ production and degradation of feed by rumen microbes. Fumarate or malate as sodium salts (Exp. 1) or acid type (Exp. 2) were added to the culture solution (150 ml, 50% strained rumen fluid and 50% artificial saliva) to achieve final concentrations of 0, 8, 16 and 24 mM, and incubated anaerobically for 0, 1, 3, 6, 9 and 12 h at $39^{\circ}C$. For both experiments, two grams of feed consisting of 70% concentrate and 30% ground alfalfa (DM basis) were prepared in a nylon bag, and were placed in a bottle containing the culture solution. Addition of fumarate or malate in both sodium salt and acid form increased (p<0.0001) pH of culture solution at 3, 6, 9 and 12 h incubations. The pH (p<0.0001) and total volatile fatty acids (VFA, p<0.05) were enhanced by these precursors as sodium salt at 3, 6 and 9 h incubations, and pH (p<0.001) and total VFA (p<0.01) from fumarate or malate in acid form were enhanced at a late stage of fermentation (9 h and 12 h) as the addition level increased. pH was higher (p<0.001) for fumarate than for malate as sodium salt at 3 h and 6 h incubations. Propionate ($C_3$) proportion was increased (p<0.0001) but those of $C_2$ (p<0.05) and $C_4$ (p<0.01 - p<0.001) were reduced by the addition of sodium salt precursors from 3 h to 12 incubation times while both precursors in acid form enhanced (p<0.011 - p<0.0001) proportion of $C_3$ from 6h but reduced (p<0.018 - p<0.0005) $C_4$ proportion at incubation times of 1, 3, 9 and 12 h. Proportion of $C_3$ was increased (p<0.05 - p<0.0001) at all incubation times by both precursors as sodium salt while that of $C_3$ was increased (p<0.001) from 6h but $C_4$ proportion was decreased by both precursors in acid form as the addition level increased. Proportion of $C_3$ was higher (p<0.01 - p<0.001) for fumarate than malate as sodium salt from 6 h incubation but was higher for malate than fumarate in acid form at 9 h (p<0.05) and 12 h (p<0.01) incubation times. Increased levels (16 and 24 mM) of fumarate or malate as sodium salt (p<0.017) and both precursors in acid form (p<0.028) increased the total gas production, but no differences were found between precursors in both chemical types. Propionate precursors in both chemical types clearly reduced (p<0.0001 - p<0.0002) $CH_4$ production, and the reduction (p<0.001 - p<0.0001) was dose dependent as the addition level of precursors increased. The $CH_4$ generated was smaller (p<0.01 - p<0.0001) for fumarate than for malate in both chemical types. Addition of fumarate or malate as sodium type reduced (p<0.004) dry matter degradation while both precursors in both chemical types slightly increased neutral detergent fiber degradability of feed in the nylon bag.

• Korean Journal of Soil Science and Fertilizer
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• v.31 no.2
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• pp.137-142
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• 1998

Assessment of N release pattern from hairy vetch (Vicia villosa Roth) residue is an important step for the efficient use of the residue as a green manure and the specific management strategy development. Objective of this research was to determine the changes of chemical composition, especially in light of the N release, when the residues of hairy vetch were applied in the soil. hairy vetch was harvested at different growth stages (April 23, May 9, and May 28) and the residues were encapsulated in the nylon mesh (1 mm) bag. The bags were placed on the surface of no-tillage plots and buried at 15cm in depth in conventional tillage plots of the corn field in early June. The bags were removed for measurements of dry matter and N content at 1, 2, 3. 5, 8 and 12 weeks after placement. Dry matter and N content of hairy vetch were higher when harvested closer to maturity, as also found in C/N ratio, cellulose and hemicellulose contents. The rate of N releases from hairy vetch residue under conventional tillage condition was greater than that under no-tillage condition. Percentages of N remaining in residue over the initial N contents under conventional tillage condition were in average 40, 23, and 15% for 1, 2, and 3 weeks after placement of mesh bag, respectively, compared with respective values of 71, 31, and 21% under no-tillage condition. The rate of N release was slower as the harvest time was closer to maturity of hairy vetch under both tillage conditions. Most of N in residue was released within 3 and 5 weeks, under conventional and no-tillage conditions, respectively. Results indicated that amounts of N released from hairy vetch residue were more closely related to the variability of N contents in residue due to the harvest time than changes of chemical composition of plant or soil management.