• Journal of the Korean Society of Food Science and Nutrition
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• v.39 no.8
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• pp.1102-1106
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• 2010

This study was carried out to investigate the effect of ethanol extract from citrus peels (CP-Et) against the alloxan-induced oxidative damage on HIT-T15, Hamster pancreatic $\beta$-cell. Total polyphenol and flavonoid contents in CP-Et were $57.00{\pm}2.91\;mg/g$ and $8.11{\pm}2.83\;mg/g$, respectively. Cell toxicity on HIT-T15 by CP-Et (0.125~0.75 mg/mL) was not observed. CP-Et (0.125 mg/mL) increased cell proliferation rate of HIT-T15, which was treated alloxan ($IC_{50}=11.58\;mM$) (cell viability=$80.52{\pm}3.29%$ of normal cell, p<0.05). In comparison with insulin secretion of oxidative damaged HIT-T15, 1.5 fold ($116.93{\pm}2.11\;{\mu}g/mg$ protein) was increased by treatment CP-Et treatment (0.125 mg/mL) in HIT-T15 (p<0.05). These results showed that CP-Et contribute to repairing cells and improvement of insulin expression on oxidative stress pancreatic $\beta$-cell, and also suggested application of CP-Et as a functional food material for type 2 diabetes.

• Applied Biological Chemistry
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• v.49 no.1
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• pp.60-64
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• 2006

The effect of far-infrared (FIR) irradiation and heat treatment on the antioxidant activity of extracts from citrus pomaces (CP) was evaluated. CP were placed in pyrex petri dishes (8.0 cm diameter) and irradiated at $100^{\circ}C$ and $150^{\circ}C$ for 5, 10, 15, 20, 40 or 60 min with a FIR heater or simple heater, respectively. After FIR irradiation or simple heat treatment, 70% ethanol extracts (EE) and water extracts (WE) of CP were prepared and radical scavenging activity (RSA) and reducing power of the extracts were determined. The antioxidant activities of the extracts increased as the time of heating or FIR-irradiation increased. When CP were FIR-irradiated at $100^{\circ}C$ far 30min, the values of RSA and reducing power of EE increased from 14.9% to 44.2%, and 0.290 to 0.886, respectively, compared to the untreated control. Simple heat treatment of CP at $100^{\circ}C$ for 60 min also increased RSA and reducing power of EE to 44.7%, and 1.045, respectively. FIR irradiation and heat treatment increased RSA and reducing power of WE from CP, too. These results indicated the antioxidant activity of CP extracts was significantly affected by FIR irradiation and heating temperature and duration on CP, and that the FIR irradiation and heat treatment process can be used as a tool for increasing the antioxidant activity of CP.

• Asian-Australasian Journal of Animal Sciences
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• v.23 no.8
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• pp.1034-1042
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• 2010

This study aimed to determine the optimum dietary crude protein level in a typical diet for fattening pigs fed ad libitum under normal climate conditions in Central Vietnam. One hundred and ninety two gilts of Mong Cai local breed (MC), $F_1$ Large White${\times}$Mong Cai and $F_2$ crossbreds of (Landrace${\times}$Mong Cai)${\times}$Large White were used. At the start of the experiment, Mong Cai pigs weighed 12 kg at 11 weeks of age, $F_1$ pigs 12.1 kg at 8 weeks of age and $F_2$ pigs 12.2 kg at 8 weeks of age. Four diets differing in crude protein (CP) content (10.1, 13.1, 16.1 and 18.9% in DM) were formulated from rice bran, corn meal, cassava meal and fish meal. Calculated digestible energy content of the diets ranged from 13.5 to 13.8 MJ per kg DM. Pigs were housed individually in pens of $2.5\;m^2$ each and had ad libitum access to feed in a trough as well as water in bowls. The final weights after a growing period of 150 days were 66, 86 and 96 kg for MC, $F_1$ and $F_2$, respectively. Feed intake of MC pigs was highest at 13.1% CP while $F_1$ and $F_2$ had the highest feed intake at 16.1% CP. The results showed that for MC the maximum gain was obtained at levels between 13 to 16% CP. For the $F_1$ the maximum gain was at dietary protein levels of 16-17%. For $F_2$ the max gain was obtained at CP levels of 16 to 18%. Feed conversion was highest in MC pigs (~4.0) followed by $F_1$ (~3.3) and $F_2$ (~3.1), and within genotypes was lowest at the optimum CP level (p<0.05). Back fat thickness in MC (33.1 mm), $F_1$ (23.0 mm) and $F_2$ (20.5 mm) pigs was different and within genotypes was the lowest at intermediate CP levels. In conclusion, increasing the dietary crude protein contents in practical diets for pigs in Vietnam can increase production on small holder farms. Optimal performance for MC, $F_1$ and $F_2$ pigs is achieved at different dietary crude protein contents.

• Journal of Microbiology and Biotechnology
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• v.26 no.1
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• pp.160-170
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• 2016

The increasing spread of antibiotic-resistant pathogens has raised the interest in alternative antimicrobial treatments. In our study, the functionally active gram-negative bacterium bacteriophage CP933 endolysin was produced in Nicotiana benthamiana plants by a combination of transient expression and vacuole targeting strategies, and its antimicrobial activity was investigated. Expression of the cp933 gene in E. coli led to growth inhibition and lysis of the host cells or production of trace amounts of CP933. Cytoplasmic expression of the cp933 gene in plants using Potato virus X-based transient expression vectors (pP2C2S and pGR107) resulted in death of the apical portion of experimental plants. To protect plants against the toxic effects of the CP933 protein, the cp933 coding region was fused at its Nterminus to an N-terminal signal peptide from the potato proteinase inhibitor I to direct CP933 to the delta-type vacuoles. Plants producing the CP933 fusion protein did not exhibit the severe toxic effects seen with the unfused protein and the level of expression was 0.16 mg/g of plant tissue. Antimicrobial assays revealed that, in contrast to gram-negative bacterium E. coli (BL21(DE3)), the gram-positive plant pathogenic bacterium Clavibacter michiganensis was more susceptible to the plant-produced CP933, showing 18% growth inhibition. The results of our experiments demonstrate that the combination of transient expression and protein targeting to the delta vacuoles is a promising approach to produce functionally active proteins that exhibit toxicity when expressed in plant cells.

• Asian-Australasian Journal of Animal Sciences
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• v.24 no.1
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• pp.65-72
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• 2011

Twenty-four Holstein dairy cows were used to evaluate the single and combined effects of different levels of crude protein (CP) and monensin treatment during early lactation on blood metabolites, milk yield and digestion of dairy cows. The experiment was designed as a completely randomized block with a $3{\times}2$ factorial arrangement of treatments. The factors were three concentrations of CP supplement (19.5, 21.4, and 23.4% of dry matter) and two levels of monensin (0 and 350 mg per cow per day). The experiment consisted of three phases and each phase was 3 wk in length. Monensin did not affect milk yield, lactose, solids-non-fat (SNF), blood glucose, triglyceride and DMI, but increased blood cholesterol, blood urea nitrogen (BUN), insulin and reduced blood ${\beta}$-hydroxybutyrate (BHBA), milk fat and protein percentage. Monensin premix significantly decreased rumen ammonia, but rumen pH and microbial protein synthesis were not affected by monensin treatment. Increasing dietary CP improved milk and protein production, but did not alter the other components of milk. Digestibility of NDF, ADF, CP were improved by increasing dietary CP. Increasing dietary CP from 19.5 to 21.4% had no significant effect on ruminal ammonia, but increasing CP to 23.4% significantly increased ruminal ammonia. There was a linear relationship between level of crude protein in the diet and volume of urine excretion. Microbial protein synthesis was affected by increasing CP level; in this way maximum protein synthesis was achieved at 23.4% CP.

• Proceedings of the Korean Society of Plant Pathology Conference
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• 2003.10a
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• pp.76.1-76
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• 2003

The coat protein (CP) of Turnip crinkle virus (TCV) is organized into 3 distinct domains, R domain (RNA-binding) connected by an arm, 5 domain and P domain. We have previously shown that the CP of TCV strongly suppresses RNA silencing, and have mapped N-terminal R domain of which is also the elicitor of resistance response in the Arabidopsis ecotype Di-17 carrying the HRT resistance gene. In order to map the region in the TCV CP that is responsible for silencing suppression, a series of CP mutants were constructed, transformed into Agrobacterium, coinfiltrated either with HC-Pro (the helper component proteinase of tobacco etch potyvirus) known as a suppressor of PTGS or GFP constructs into leaves of Nicotiana benthmiana expressing GFP transgenically. In the presence of HC-Pro, all CP mutants were well protected, accumulating mutant CP mRNAs and their proteins even 5 days post-infiltration (DPI). In the presence of GFP, some mutant constructs which showed the accumulation of CP mutants and GFP mRNAs at early stage but eventually degraded at 5 DPI. Only a mutant which carrying 4 amino acid deletion of R domain was tolerable to maintain suppressing activity, suggesting that the suppressing activity is not directly related with the eliciting activity. A transient assay also revealed that the mutants synthesized their proteins, suggesting that a full length of CP sequences and its intact structure are required to stabilize CP, which suppresses the RNA silencing.

• BMB Reports
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• v.40 no.4
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• pp.486-493
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• 2007

Atopic dermatitis (AD) is a chronic inflammatory skin disease and the pathogenesis of AD is associated with the release of various cytokines/chemokines due to activated $Th_2$ immune responses. Synthetic oligodeoxynucleotides (ODNs) containing unmethylated CpG dinucleotide in the context of particular base sequence (CpG motifs) are known to have the immunostimulatory activities in mice and to convert from Th2 to Th1 immune responses in AD. We aimed to investigate that CpG ODN, especially phosphodiester form, can stimulate the protective immunity in NC/Nga mice with AD. We isolated BMDCs from NC/Nga mice and then, cultured with GM-CSF and IL-4 for 6 days, and treated for 2 days by either phosphorothioate ODN or phosphodiester ODN. CpG ODN-treated DCs resulted in more production of IL-12. When CpG ODN-treated DCs were intravenously injected into the NC/Nga mice, the NC/Nga mice with CpG ODN-treated DCs showed significant improvement of AD symptoms and decrease of IgE level. Histopathologically, the NC/Nga mice skin with CpG ODN-treated DCs showed the decreased IL-4 and TARC expression comparing with non-injected mice. These results may suggest that phosphodiester CpG ODN-treated DCs might function as a potent adjuvant for AD in a mouse model.

• Asian-Australasian Journal of Animal Sciences
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• v.13 no.11
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• pp.1523-1528
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• 2000

Four Suffolk ewes were used in Latin Square switch over design to study the effects of varying levels and sources of protein on heat production and thermoregulatory responses at daytime high ($33^{\circ}C$ temperature. They were fed Italian ryegrass hay supplemented with fishmeal and/or urea, providing three different levels of crude protein (CP) (low/unsupplemented: 7.9, medium: 11.6, and high: 15.8%) at $1.5{\times}maintenance$. Feeds were distributed at 0900 (30%) and 1700 (70%). Urea diet caused higher heat production and increased vaginal temperature compared to fishmeal and fishmeal-urea mix diets. Time spent standing, skin temperature and respiration rate of sheep fed urea were similar with those of sheep fed fishmeal. Sheep fed diet with low CP level had higher heat production, increased vaginal and skin temperature than sheep fed diet with medium CP content. Sheep on high CP diet produced significantly more heat than sheep fed medium CP diets. Their vaginal temperatures were similar with those of sheep fed medium CP diet but lower than those of sheep fed low CP diet. Respiration rates of sheep and time spent by them for standing on all diets did not differ significantly. These results suggest that urea is inferior protein supplement for thermoregulation of animal at hot environment, as it induced higher heat production than fishmeal and fishmeal-urea mix. Thermoregulatory response on fishmeal-urea mix diet was similar to fishmeal diet. Increasing CP of the diet from low to medium gives advantage for thermoregulation of animal. Increasing CP further to high level was not beneficial as it resulted in the responses of sheep similar to those on low protein diet.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.6
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• pp.2485-2489
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• 2014

In the present study, we studied the hypermethylation of the human riboflavin transporter 2 (hRFT2) gene and regulation of protein expression in biopsies from resected tissues from Uighur cervical squamous cell carcinoma (CSCC) patients and their neighboring normal tissues. hRFT2 gene promoter region methylation sequences were mapped in cervical cancer cell line SiHa by bisulfite-sequencing PCR and quantitative detection of methylated DNA from 30 pairs of Uighur's CSCCs and adjacent normal tissues by MassARRAY (Sequenom, San Diego, CA, USA) and hRFT2 protein expression was analyzed by immunohistochemistry. In SiHa, we identified 2 CG sites methylated from all of 12CpG sites of the hRFT2 gene. Analysis of the data from quantitative analysis of single CpG site methylation by Sequenom MassARRAY platform showed that the methylation level between two CpG sites (CpG 2 and CpG 3) from CpG 1~12 showed significant differences between CSCC and neighboring normal tissues. However, the methylation level of whole target CpG fragments demonstrated no significant variation between CSCC ($0.476{\pm}0.020$) and neighboring normal tissues ($0.401{\pm}0.019$, p>0.05). There was a tendency for translocation the hRFT2 proteins from cytoplasm/membrane to nucleus in CSCC with increase in methylation of CpG 2 and CpG 3 in hRFT2gene promoter regions, which may relate to the genesis of CSCC. Our results suggested that epigenetic modifications are responsible for aberrant expression of the hRFT2 gene, and may help to understand mechanisms of cervical carcinogenesis.

• Asian-Australasian Journal of Animal Sciences
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• v.14 no.12
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• pp.1728-1733
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• 2001

An experiment was conducted at the experimental duck farm of Cantho University in the Mekong Delta of Vietnam to evaluate the effects of offering pairwise feeds with different crude protein (CP) levels to Cherry Valley broiler ducks from 28-63 days of age on feed intake and performance. Five diets consisting of mixtures of soya bean meal, fish meal, maize meal and fine rice bran, were formulated to give isocaloric diets with 30, 24, 18, 16 and 12% CP. These were offered separately as pellets to growing ducklings in the following combinations: 30+18, 30+12, 24+16 and 18+12. A diet with 20% CP (CP20) was used as control. All the diets were offered ad libitum. Daily feed intakes (g dry matter, DM) were 161, 168, 150, 101 and 143 g for the 30+18, 30+12, 24+16, 18+12 and CP20 treatments, respectively. CP intakes were 29.6, 27.0, 24.8, 17.1 and 22.9% of total DM intake, and the ratio of methionine+cystine to lysine was 0.76, 0.75, 0.75, 0.76 and 0.75 for the 30+18, 30+12, 24+16, 18+12 and CP20 treatments, respectively. Live weights at 63 days of age were 2,937, 3,054, 2,898, 2,200, and 2,811 g (p<0.001), and daily live weight gains 57.8, 61.7, 57.0, 38.5 and 55.7 g (p<0.001) for the 30+18, 30+12, 24+16, 18+12 and CP20 treatments, respectively. The weights and yields of the carcass, breast and thigh muscles, and heart decreased in the order 30+18, 30+12, 24+16, CP20 and 18+12 (p<0.001). The present study suggests that growing broiler offered a combination of high and low protein feeds pairwise will eat excessive amounts of the high protein feed, thus resulting in protein intakes above requirements.