• Journal of Microbiology and Biotechnology
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• v.6 no.6
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• pp.474-481
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• 1996

Since the commercially available rabbit anti-cyclin D3, generated from c-terminal 16 amino acid residues which are common to human and murine cyclin D3, is highly cross-reactive with many other cellular proteins of mouse, a new rabbit polyclonal anti-cyclin D3 has been raised by using murine cyclin D3 protein expressed at a high level in Escherichia coli as the immunogen. To express murine cyclin D3 protein in E. coli, the cyclin D3 cDNA fragment encoding c-terminal 236 amino acid residues obtained by polymerase chain reaction (PCR) was inserted into the NcoI/BamHI site of protein expression vector, pET 3d. Molecular mass of the cyclin D3 overexpressed in the presence of IPTG (Isopropyl $\beta$-D-thiogalactopyranoside) was approximately 26 kDa as calculated from the reading frame on the DNA sequence, and the protein was insoluble and mainly localized in the inclusion bodies that could be easily purified from the other cellular soluble proteins. When renaturation was performed following denaturation of the insoluble cyclin D3 protein in the inclusion bodies using guanidine hydrochloride, 4.4 mg of soluble form of cyclin D3 protein was produced from the transformant cultured in 100ml of LB media under the optimum conditions. Four-hundred micrograms of the soluble form of cyclin D3 protein was used for each immunization of a rabbit. When the antiserum obtained 2 weeks after tertiary immunization was applied to Western blot analysis, it was able to detect 33 kDa cyclin D3 protein in both murine lymphoma cell line BW5147.G.1.4 and human Jurkat T cells at 3,000-fold dilution with higher specificity to murine cyclin D3, demonstrating that the new rabbit polyclonal anti-murine cyclin D3 generated against c-terminal 236 amino acid residues more specifically recognizes murine cyclin D3 protein than does the commercially available rabbit polyclonal antibody raised against c-terminal 16 amino acids residues.

• Microbiology and Biotechnology Letters
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• v.32 no.1
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• pp.72-77
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• 2004

Previously, we synthesized a novel Cyclin-dependent kinase inhibitor, MCS-5A. Also, we investigated the involvement of cell cycle regulatory events during MCS-5A-mediated apoptosis in HL-60(+p16/-p53) cells with up-regulation of p16 protein expression. In contrast, apoptosis was not observed in A549(-p16/+p53) cells. Therefore we propose that $p16^{INK4A}$ is a key enzyme for inducing apoptosis. In the present studies, we have explored the mechanism of $p16^{INK4A}$ -mediated cytotoxicity and the role of p16.sup INK4A/ overexpression in the induction of apoptosis in human tumor cells. The tumor suppressor gene $p16^{INK4A}$ is known as a cyclin-dependent kinase inhibitor (CKI) and cell cycle regulator. We expressed wild type $p16^{INK4A}$ in pcDNA3.1 vector and then transfected into non-small cell lung cancer (NSCLC) cell expressing different statue of p16$^{INK4A}$, p53 gene〔A549(-p16/+p53), H1299(-p16/-p53) and HeLa(+pl6/+p53) cell line〕. TUNEL assay (including propidium iodide staining following transfection of these cell line with pcDNA3.1-pl6) indicate that p16$^{INK4A}$-mediated cytotoxicity was associated with apoptosis. This is supported by studies demonstrating an induction of caspase 3 cleavage due to the transfection of A549, H1299 and HeLa cells with pcDNA3.1-pl6. These results suggest that p16$^{INK4A}$ has a new function of inducing apoptosis which is not related with the function of tumor suppressor gene p53.

• Applied Biological Chemistry
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• v.39 no.3
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• pp.177-182
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• 1996

Effects of 14 carbohydrates supplied as carbon sources on cell growth and sporulation of, and the production of insecticidal crystal proteins by Bacillus thuringiensis strains were investigated in liquid cultures. Strains grew well in media containing any one of the 14 carbohydrates supplied, reaching maximum cell densities of $10^7{\sim}10^8\;cells/ml$ in 16.7 to 22 hours after inoculation depending on the strain. Spores first appeared in 16.7 to 24.7 hours after inoculation, and 80% sporulation was reached in 28 to 51.3 hours after inoculation depending on the strain. No change in pH of media was observed after cell multiplication. The production of total protein was highest when supplied with sucrose and was lowest with starch. More insecticidal crystal proteins were produced when supplied with glucose, lactose, maltose, or sucrose. The amount of insecticidal crystal proteins produced by the strains was proportional to that of the total protein. The relative amount of individual insecticidal crystal protein species produced by B.t. kurstaki and B.t. israelensis was not influenced by the carbohydrates supplied.

• Proceedings of the Korean Society of Crop Science Conference
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• 2022.10a
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• pp.301-301
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• 2022

Dairy products are important diet source for human because of their balanced essential nutrients along with various vitamins and minerals. However, lactose in milk can result in diarrhea to some consumers with lactose intolerance. Soy milk has no lactose and is suitable as a substitute for diary milk in accordance with recent trend of replacing animal food with vegetable food. However, polysaccharides in soy milks are difficult for humans to digest, leading to flatulence. These polysaccharides can be decomposed into monosaccharides by lactic acid bacteria, and fermentation can improve food quality. Because mungbean has higher carbohydrate content than soybean, mung milk can be easily fermented than soy milk, resulting in vege yogurt with higher contents of lactic acid. In this study, fermentation characteristics of vege yogurt were analyzed with different ratio of soy milk and mung milk (0%, 25%, 50%, 75%, 100% and 0%+sucrose) and different fermentation time (0, 8, and 16 hours). In general, pH decreased as fermentation time increased. Overall, pH significantly decreased when the mung milk content in yogurt increased. All samples showed higher titratable acidity after fermentation and soy yogurt (mungbean 0%, 16 hours) with sucrose showed the highest value (6.825%). As fermentation time increase, viscosity increased. In 8 and 16 hours, mung milk yogurt (mungbean 100%) showed the lowest viscosity while soy milk yogurt (soybean 100%) with no sucrose showed the highest viscosity after 16 hours of fermentation. The contents of crude protein, crude fat and ash were measured for nutritional analysis. Soy milk (mungbean 0%, 0 hours) had the values of crude protein 2.9g, crude fat 1.8g, and ash 0.3g, and mung milk (mungbean 100%, 0 hours), showed the values of crude protein 1.7g, crude fat 0g, and ash 0.3g. To analyze the effect of the differences in the contents of nutrition between soy milk and mung milk on fermentation, the changes in sugar contents, and antioxidant capacity will be conducted depending on fermentation time. Our results will provide the information in researching plant beverages.

• Journal of Life Science
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• v.18 no.2
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• pp.193-199
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• 2008

The purpose of this study was to demonstrate the effectiveness of walking and band exercise for 12 weeks on c-reactive protein and cardiovascular disease risk factor in overweight and obese children. Body composition, blood lipids, insulin sensitivity, c-reactive protein (CRP) were assessed before, after 4 weeks and after 12 weeks of combined exercise. Sixteen participants (BMI${\geq}$21.3) were randomly allocated to exercise group (n=8) and control group (n=8). The exercise group participated in 50 minutes of walking exercise and band exercises as resistance training two days a week for 12 weeks. There were significant different on weight (p<0.001), BMI (p<0.001), fat mass (p<0.001), fat percentage (p<0.001), LBM percentage (p<0.001), TG (p<0.05), HDL-C (p<0.01), insulin (p<0.05), HOMA-IR (p<0.05) in exercise group after intervention. And the change of weight (p<0.001), BMI (p<0.001), fat mass (p<0.001), fat percentage (p<0.001), LBM mass (p<0.05), LBM percentage (p<0.001), insulin (p<0.05), HOMA-IR (p<0.05) were significant difference between groups after intervention. These findings suggest that 12 weeks of walking and band exercise can be useful intervention in the improvement of cardiovascular disease risk factor in overweight and obese children. But c-reactive protein was no change.

• Asian-Australasian Journal of Animal Sciences
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• v.32 no.4
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• pp.556-563
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• 2019

Objective: This experiment was conducted to investigate the effect of reducing dietary metabolic energy (ME) and crude protein (CP) levels on growth performance, blood profiles, and nutrient digestibility in weaning pigs. Methods: A total of 240 crossbred pigs ($Duroc{\times}[Landrace{\times}Yorkshire]$) with an average body weight of $8.67{\pm}1.13kg$ were used for a 6-week feeding trial. Experimental pigs were allotted to a $2{\times}3$ factorial arrangement using a randomized complete block design. The first factor was two levels of dietary ME density (low ME level, 13.40 MJ/kg or high ME level, 13.82 MJ/kg) and the second factor was three dietary CP levels based on subdivision of early and late weaning phases (low CP level, 19.7%/16.9%; middle CP level, 21.7%/18.9%; or high CP level, 23.7%/20.9%). Results: Over the entire experimental period, there were no significant difference in body weight among groups, but a decrease in diet energy level was associated with an increase in average daily feed intake (p = 0.02) and decrease in gain-feed ratio (G:F) ratio (p<0.01). Decreased CP levels in the diet were associated with a linear increase in average daily gain (p<0.05) and quadratic increase in G:F ratio (p<0.05). In the early weaning period, blood urea nitrogen concentration tended to increase when ME in the diet decreased and decrease when CP level in the diet decreased (p = 0.09, p<0.01, respectively). Total protein concentration tended to increase when CP level was reduced (p = 0.08). In the late weaning period, blood urea nitrogen concentration decreased linearly as CP level decreased (p<0.01). The CP and crude fat digestibility decreased when ME was decreased by 0.42 MJ/kg (p = 0.05, p = 0.01, respectively). The CP digestibility increased linearly as CP level decreased (p = 0.01). Conclusion: A weaning pig diet containing high ME level (13.82 MJ/kg) and low CP level (19.7%/16.9%) can improve pig growth performance and nutrient digestibility.

• Molecular & Cellular Toxicology
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• v.2 no.1
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• pp.54-59
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• 2006

The toxicity of cadmium on Caenorhabditis elegans was investigated to identify sensitive biomarkers for environmental monitoring and risk assessment. Stress-related gene expression were estimated as toxic endpoints Cadmium exposure led to an increase in the expression of most of the genes tested. The degree of increase was more significant in heat shock protein-16.1, metallothionein-2, cytochrome p450 family protein 35A2, glutathione S-transferase-4, superoxide dismutase-1, catalase-2, C. elegans p53-like protein-1, and apoptosis enhancer-1 than in other genes. The overall results indicate that the stress-related gene expressions of C. elegans have considerable potential as sensitive biomarkers for cadmium toxicity monitoring and risk assessment.

• Asian-Australasian Journal of Animal Sciences
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• v.14 no.4
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• pp.525-534
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• 2001

A total of 120 pigs were used to investigate the effects of yucca extracts on the growth performance, nutrient digestibility, nutrient excretion and carcass characteristics of finishing pigs fed different levels of dietary protein. Pigs were allotted into $2{\times}3$ factorial design by the supplementation of yucca extracts (YE, 0 and 120 mg/kg) and 3 levels of dietary protein (14, 16, 18% for early finisher and 12, 14, 16% for late finisher for low, medium and high protein diet, respectively). During the early finishing period (51~76 kg BW), no significant difference was found in growth performance regardless of the YE supplementation or dietary protein levels. Growth performance of late finishing pigs (76~101 kg BW) was also not significantly different among treatments. However, ADG of pigs fed YE diet was significantly improved (p<0.05) regardless of the dietary protein levels. For the overall period (51~101 kg BW), although adding YE to the diet and elevating the protein level showed better ADG, there were no significant differences on growth performance among treatments. Early finishers showed significantly higher crude protein, crude ash and crude fat digestibilities when they were fed diets supplemented with YE. Digestibilities of amino acids were not affected by YE. Late finishers did not show any significant differences in proximate nutrient digestibilities regardless of YE supplementation or dietary protein levels. YE tended to slightly improve the CP digestibility, however no significant difference was found with increased dietary protein levels. There was no significant difference in amino acid digestibilities with YE supplementation or dietary CP levels during the late finishing period. Dry matter (DM) and nitrogen (N) excretion in feces did not show any significant difference among treatments. Early finishing pigs also did not respond to the inclusion of YE or dietary protein levels (p<0.05). Fecal N excretion of early finishing pigs seemed to be lowered in pigs fed YE. Pigs fed medium dietary protein diet tended to excrete a higher amount of N during the early finishing period, but not statistically different. A slight increase in fecal N excretion was found with the increased level of dietary protein during the late finishing period. For ammonia nitrogen excretion, although there was no significance, the NH3-N content tended to be increased by the increased dietary protein levels and with YE supplementation. The NH3-N content in manure increased by 24.5% with YE supplementation. There were no significant differences in carcass weight, backfat thickness, carcass grade and loin eye area among treatments. However, pigs fed non-YE with low protein diet showed a significantly (p<0.05) low carcass ratio among treatments and there was significant (p<0.05) difference between the YE-added treatment and non YE treatment in carcass ratio. As for the feed cost, the cost of feeding high level protein was higher than that of medium level protein by 5% and low level protein by 9% (p<0.05). Therefore, based on this study, it could be concluded that environmentally friendly agents might play a role to some extent in finishing pigs from the aspect of pollution control, and that more than 14 and 12% of dietary protein for early finishing and late finishing pigs respectively do not necessarily guarantee high growth performance.

• Asian-Australasian Journal of Animal Sciences
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• v.27 no.9
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• pp.1303-1310
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• 2014

A 12 week growth study was carried out to investigate the supplemental effects of dietary garlic powder (GP) on growth, feed utilization and whole body composition changes of fingerling sterlet sturgeon Acipenser ruthenus (averaging weight, 5.5 g). Following a 24-h fasting, 540 fish were randomly distributed to each of 18 tanks (30 fish/tank) under a semi-recirculation freshwater system. The GP of 0.5% (GP0.5), 1% (GP1), 1.5% (GP1.5), 2% (GP2) and 3% (GP3) was added to the control diet (GP0) containing 43% protein and 16% lipid. After the feeding trial, weight gain (WG) of fish fed GP1.5, GP2 and GP3 were significantly higher (p<0.05) than those of fish fed GP0, GP0.5 and GP1. Feed efficiency and specific growth rate (SGR) showed a similar trend to WG. Protein efficiency ratio of fish fed GP1.5, GP2, and GP3 were significantly higher (p<0.05) than those of fish groups fed the other diets. A significant difference (p<0.05) was found in whole body composition (moisture, crude protein, crude lipid, ash, and fiber) of fish at the end of the experiment. Significantly higher (p<0.05) protein and lipid retention efficiencies (PRE and LRE) were also found in GP1.5, GP2, and GP3 groups. Broken-line regression model analysis and second order polynomial regression model analysis relation on the basis of SGR and WG indicated that the dietary optimal GP level could be greater than 1.77% and 1.79%, but less than 2.95% and 3.18% in fingerling sterlet sturgeon. The present study suggested that dietary GP for fingerling sterlet sturgeon could positively affect growth performance and protein retention.

• Asian-Australasian Journal of Animal Sciences
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• v.30 no.3
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• pp.386-391
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• 2017

Objective: Protein supplementation is costly and can result in excess nitrogen (N) excretion. The objective of this study was to evaluate the effects of feeding different levels of dietary protein on average daily gain, body size, rumen fermentation, and nitrogen excretion of 8 to 10 month-old Holstein heifers. Methods: Thirty-six Holstein heifers were divided into 12 blocks according to age ($273{\pm}6.2d$) and were randomly assigned to diets containing a low (10.2% dry matter [DM]), medium (11.9% DM), or high (13.5% DM) level of dietary crude protein (CP). All diets contained approximately 70% roughage and 30% concentrate with similar dietary metabolizable energy (ME) content (2.47 Mcal/kg). Results: Dry matter intake did not differ among the treatments, and average daily gain increased with the increasing dietary protein, 0.79, 0.95, 0.97 kg/d for low, medium, and high group, respectively. Body height increased linearly with increasing dietary CP but no other significant differences in body dimensions were found among the treatments. The increased ratios of dietary CP improved the rate of rear teat length growth remarkably (p<0.05). There was no difference in rumen pH or ruminal major volatile fatty acid (acetate, propionate, and butyrate) concentration among the 3 diets, but rumen ammonia-N concentration increased with the higher dietary CP (p<0.05). Increasing N intake led to increased total N excretion; urinary N excretion was significantly increased (p<0.05) but fecal N excretion was similar among the treatments. Conclusion: These data suggest that the diet containing 11.9% CP (ME 2.47 Mcal/kg) could meet the maintenance and growth requirements of 9 to 11 month-old Holstein heifers gaining approximately 0.9 kg/d.