• Korean Journal of Poultry Science
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• v.50 no.1
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• pp.31-39
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• 2023

The objective of this study was to investigate the effect of exogenous protease supplementation in diets formulated with poultry by-product meal on growth performance, small intestine magnitudes, and meat physiochemical characters in broiler chickens from 21 to 35 days post-hatch. A total of 120, one-day-old "Arbor Acres Plus" broiler chickens (male: female - 50:50) were allocated one of two dietary treatments to give six replicates and ten birds per cage. Two dietary treatments included a control diet (CON) and a diet supplemented with protease (CON+Pro). At day 35, body weight and feed intakes were measured to calculate the feed conversion ratio for the entire experiment period. Two birds from each pen were euthanized to measure the relative lengths and relative weights of three small intestine sections and meat samples were collected for physiochemical characteristic analyses at 35 days post-hatch. Exogenous protease supplementation did not influence (P>0.05) growth performance but showed a tendency to improve FCR (P=0.082). Protease supplementation showed a tendency to reduce proximal small intestine length (P=0.091). Broilers offered dietary treatments influenced minced meat color where protease supplementation resulted in lower CIE b^* (P<0.001) colorimetric value for yellowness and showed a significant trend (P=0.059) on reducing meat redness CIE a^*. In conclusion, the addition of exogenous protease to a broiler diet formulated with poultry by-product meal did not affect the growth performance, small intestine magnitudes, and meat physiochemical parameters (except CIE b^*) in broiler chickens.

• Journal of Life Science
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• v.21 no.2
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• pp.309-315
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• 2011

Rice syrup meal (RSM) was enzymatically hydrolyzed using eight commercial proteases (Protamex, Neutrase, Flavourzyme, Alcalase, Protease M, Protease N, Protease A, Molsin F) for 4 hr at optimum pH and temperature. Proteolytic hydrolysates were examined in supernatant and precipitate using Lowry protein assay, semimicro Kjeldahl method and gravimetric method using weight difference before and after enzymatic hydrolysis. Although RSM contains a high amount of protein (71.2%), only a very small amount of protein was hydrolyzed. Two proteases (Protease M and Protease N) were found to be the most effective in the hydrolysis of RSM protein. In Lowry method, 57.5 and 59.0 mg protein/g RSM were hydrolyzed after Protease M and Protease N treatments, respectively. In gravimetric method, 80.0 and 85.4 mg protein/g RSM were hydrolyzed after Protease M and Protease N treatments. In Kjeldahl method, 67.43 and 70.43 mg protein/g RSM were hydrolyzed after Protamex and Protease N treatments, respectively. For synergistic effect, two or three effective commercial proteases (Protease M, Protease N and Protease A) were applied to RSM at one time. The highest hydrolysis of RSM protein was observed in both Lowry protein assay (80.3 mg protein/g RSM) and gravimetric methods (153.2 mg protein/g RSM) when three commercial proteases were applied at one time, suggesting the synergistic effect of those proteases.

• Korean journal of food and cookery science
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• v.28 no.1
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• pp.41-49
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• 2012

This purpose of this study was to develop a tea with small red bean which have been known to have effect regarding anti-obesity, fatigue recovery, edema recovery, blood circulation, etc. In order to provide baseline data for small red bean tea we investigated the general components, antioxidative effect and proanthocyanidin analysis in small red beans[Phaseolus angularis W.F. Wight.]. Physicochemical analysis(pH, sugar content, salinity, turbidity), color, anti oralmicrobial activity, content of saponin and sensory test of small red bean with different boiling time in 8 minutes(SR1), 16 minutes(SR2), 24 minutes(SR3), 32 minutes(SR4), 40 minutes(SR5) was also measured. It was shown that the crude fat, carbohydrate, moisture, crude protein, crude ash content of small red bean were 1.0%, 63.9%, 12.8%, 18.7%, 3.6%. DPPH free radical scavenging activity, the total phenolic compounds content and flavonoid content increased significantly (p<0.001). The results of analyzed proanthocyanidin was distinguished by characteristic UV-visible spectra with absorption maximum at 320 nm($t_R$ 7.589 min). As the boiling time(8 minutes:SR1, 16 minutes:SR2, 24 minutes:SR3, 32 minutes:SR4, 40 minutes:SR5) of small red beans increaseds, the pH significantly decreased(p<0.001). The sugar content, salinity and turbidity significantly increased(p<0.001). Moreover, Hunter L, a and b values, crude saponin also increased(p<0.001). The results of analyzed activity against oral bacteria, S. mutans, S. sobrinus, P. intermedia and P. gingivalis showed a higher antibacterial activities than E. coli and S. aureus. MIC was measured that S. mutans, S. sobrinus, P. intermedia and P. gingivalis showed a lower MICs than E. coli and S. aureus. The results regarding sensory test measures, In case of color, refreshing taste and overall quality, SR3 had the highest preference overall among tested samples. In cases conceming odor and taste, SR5 had the highest preference and with regards to sweetness and saltyness, SR4 had the highest preference.

• Asian-Australasian Journal of Animal Sciences
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• v.24 no.5
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• pp.670-677
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• 2011

The growth rate of the young pig is generally much less than it's potential and may be constrained by endocrine status as well as nutrient intake. Growth factors are present in relatively high quantities in colostrum and play an important part in gut development. It is possible that supplementation of colostrum protein isolate may stimulate gut and whole body growth in the pig. Eight male and 8 female (Large Whitex${\times}$Landrace) piglets were weaned at 1 d of age after each pig had obtained colostrum from their dam, and were trained to consume one of two liquid diets. The two diets were based on either a colostrum protein isolate (n = 4 males and 4 females) or whey protein concentrate (n = 4 males and 4 females) and were formulated to contain equal levels of crude protein and amino acids. Pigs were fed their diets ad libitum for 28 days after which time 12 pigs were euthanised and various tissues and organs weighed. Pigs were bled for IGF-I analyses at 21 and 28 days of age. Daily gain was higher in pigs consuming the colostrum isolate (171 vs. 216 g/d, p = 0.010), particularly between 2 and 4 weeks of age (212 vs. 298 g/d, p = 0.010). Pigs tended to consume more of the liquid feed containing colostrum isolate (25.5 vs. 29.1 kg, p = 0.074) and gained more live weight per unit of liquid feed (0.203 vs. 0.223 g/g, p = 0.056). There were no effects of sex on growth performance. Pigs consuming the diet supplemented with colostrum isolate had higher (p<0.05) full gut weight (445 vs. 554 g, p = 0.026), empty gut weight (356 vs. 463 g, p = 0.008), stomach weight (42.2 vs. 54.4 g, p = 0.001), small intestine weight (222 vs. 275 g, p = 0.025) and large intestine weight (63.7 vs. 98.0 g, p = 0.005). Plasma IGF-I (99 vs. 150 ng/ml, p<0.001) and IGF-II (265 vs. 406 ng/ml, p<0.001) were higher in pigs fed colostrum isolate. Pigs consuming colostrum protein isolate ate more, grew faster and had higher plasma IGF-I concentrations than pigs consuming a diet with similar macronutrient content but devoid of growth factors.

• Archives of Plastic Surgery
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• v.41 no.6
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• pp.654-660
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• 2014

Background Reactive oxygen species (ROS) damages cell molecules, and modifies cell signaling. The nuclear factor E2-related factor (Nrf2) is a critical transcription regulator, which protects cells against oxidative damage. Nrf2 expression is increased in a large number of cancers. However, little information has been reported regarding the expression of Nrf2 in skin cancers. Hence, we explored the expression of Nrf2 protein in skin cancers. Methods The Nrf2 protein expression in 24 specimens, including 6 malignant melanomas (MM), 6 squamous cell carcinomas (SCC), 6 basal cell carcinomas (BCC), and 6 normal skin tissues, was evaluated by western blotting. Immunohistochemical staining was performed. The expression of Kelch-like ECH-associated protein 1 (Keap1), the key regulator of Nrf2, was also analyzed by western blotting. Results Small interfering RNA transfection to the melanoma cell line G361 confirmed that an approximately 66 kDa band was the true Nrf2 band. The western blot revealed that the Nrf2 protein was definitely expressed in normal skin tissues, but the Nrf2 expression was decreased in MM, SCC, and BCC. Immunohistochemical examination showed that expression of Nrf2 was decreased in all skin cancer tissues compared to the normal skin tissues. Keap1 was not expressed in all malignant skin tumors and normal skin tissues by western blot. Conclusions ROS was increased in various types of cancers which proteins were highly expressed or underexpressed. This study demonstrated that the expression of Nrf2 protein was down-regulated in human malignant skin tumors. We suggest that decreased expression of Nrf2 is related to skin cancers.

• The Korean Journal of Physiology and Pharmacology
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• v.20 no.3
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• pp.315-324
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• 2016

Human cardiac fibroblasts (HCFs) have various voltage-dependent $K^+$ channels (VDKCs) that can induce apoptosis. Hydrogen peroxide ($H_2O_2$) modulates VDKCs and induces oxidative stress, which is the main contributor to cardiac injury and cardiac remodeling. We investigated whether $H_2O_2$ could modulate VDKCs in HCFs and induce cell injury through this process. In whole-cell mode patch-clamp recordings, application of $H_2O_2$ stimulated $Ca^{2+}-activated$ $K^+$ ($K_{Ca}$) currents but not delayed rectifier $K^+$ or transient outward $K^+$ currents, all of which are VDKCs. $H_2O_2-stimulated$ $K_{Ca}$ currents were blocked by iberiotoxin (IbTX, a large conductance $K_{Ca}$ blocker). The $H_2O_2-stimulating$ effect on large-conductance $K_{Ca}$ ($BK_{Ca}$) currents was also blocked by KT5823 (a protein kinase G inhibitor) and 1 H-[1, 2, 4] oxadiazolo-[4, 3-a] quinoxalin-1-one (ODQ, a soluble guanylate cyclase inhibitor). In addition, 8-bromo-cyclic guanosine 3', 5'-monophosphate (8-Br-cGMP) stimulated $BK_{Ca}$ currents. In contrast, KT5720 and H-89 (protein kinase A inhibitors) did not block the $H_2O_2-stimulating$ effect on $BK_{Ca}$ currents. Using RT-PCR and western blot analysis, three subtypes of $K_{Ca}$ channels were detected in HCFs: $BK_{Ca}$ channels, small-conductance $K_{Ca}$ ($SK_{Ca}$) channels, and intermediate-conductance $K_{Ca}$ ($IK_{Ca}$) channels. In the annexin V/propidium iodide assay, apoptotic changes in HCFs increased in response to $H_2O_2$, but IbTX decreased $H_2O_2$-induced apoptosis. These data suggest that among the VDKCs of HCFs, $H_2O_2$ only enhances $BK_{Ca}$ currents through the protein kinase G pathway but not the protein kinase A pathway, and is involved in cell injury through $BK_{Ca}$ channels.

• Asian Pacific Journal of Cancer Prevention
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• v.14 no.2
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• pp.685-689
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• 2013

Objective: To study the effects of down-regulation of HDAC6 expression on proliferation, cell cycling and migration of esophageal squamous cell carcinoma (ESCC) cells and related molecular mechanisms. Methods: ESCC cell line EC9706 cells were randomly divided into untreated (with no transfection), control siRNA (transfected with control siRNA) and HDAC6 siRNA (transfected with HDAC6 small interfering RNA) groups. Effects of HDAC6 siRNA interference on expression of HDAC6 mRNA and protein in EC9706 cells were investigated by semi-quantitative RT-PCR, Western blotting and immunocytochemistry methods. Effects of down-regulation of HDAC6 expression on cell proliferation, cell cycle, and cell migration were studied using a CCK-8 kit, flow cytometry and Boyden chambers, respectively. Changes of mRNA and protein expression levels of cell cycle related factor (p21) and cell migration related factor (E-cadherin) were investigated by semi-quantitative RT-PCR and Western blotting methods. Results: After transfection of HDAC6 siRNA, the expression of HDAC6 mRNA and protein in EC9706 cells was significantly downregulated. In the HDAC6 siRNA group, cell proliferation was markedly inhibited, the percentage of cells in G0/G1 phase evidently increased and the percentage of cells in S phase decreased, and the number of migrating cells significantly and obviously decreased. The mRNA and protein expression levels of p21 and E-cadherin in the HDAC6 siRNA group were significantly higher than those in the untreated group and the control siRNA group, respectively. Conclusions: HDAC6 siRNA can effectively downregulate the expression of HDAC6 mRNA and protein in EC9706 cells. Down-regulation of HDAC6 expression can obviously inhibit cell proliferation, arrest cell cycling in the G0/G1 phase and reduce cell migration. The latter two functions may be closely related with the elevation of mRNA and protein expression of p21 and E-cadherin.

• Journal of Life Science
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• v.34 no.7
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• pp.493-499
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• 2024

Antimicrobial peptides are antimicrobial substances inherent in animals and plants, with strong antibacterial activity even in small amounts and with various other functions such as antiviral and antioxidant actions. Plants can be grown with just water and sunlight, allowing for their mass production at low costs. However, transforming a chloroplast into one that produces antimicrobial peptides, rather than growing plants, increases the amount of protein expression and minimizes contamination of the ecosystem because gene transfer by pollen does not occur. In that context, using transgenic plant chloroplasts to produce recombinant proteins increases protein degradation and reduces the solubility of proteins. To solve this problem, we fused SUMO, a fusion protein, with a recombinant protein. We also used a 6xHis tag to purify the fusion protein. The antimicrobial peptide stomoxyn is an antibacterial substance found in stable flies. Stomoxyn has an α-helix structure and is amphiphilic, which allows it to dissolve bacterial cell membranes. In this study, we constructed a transformation vector to express stomoxyn in both plant chloroplasts and Escherichia coli and used this vector to confirm the expression of stomoxyn in E. coli. The expression of the protein was then confirmed in E. coli using a transformation vector. The expressed stomoxyn was purified by nickel column and SUMOase treatment, and its antibacterial activity was confirmed using an agar diffusion assay. The EGFP gene was used to ensure that the transformed vector was inserted into the chloroplast.

• Tuberculosis and Respiratory Diseases
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• v.44 no.4
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• pp.796-805
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• 1997

Background : It is clear that deregulation of cell cycle progression is a hallmark of neoplastic transformation and genes involved in the $G_1$/S transition of the cell cycle are especially frequent targets for mutations in human cancers, including lung cancer. p16 gene product, one of the G1 cell-cycle related proteins, that is recently identified plays an important role in the negative regulation of the the kinase activity of the cyclin dependent kinase (cdk) enzymes. Therefore p16 gene is known to be an important tumor suppressor gene and is also called MTS1 (multiple tumor suppressor 1). No more oncogenes have been reported to be frequently related to multiple different malignancies than the alterations of p16 gene. Especially when it comes to non-small cell lung cancer, there was no expression of p16 in more than 70% of cell lines examined. And also it is speculated that p16 gene could exert a key role in the development of non-small cell lung cancer. This study was designed to evaluate whether p16 gene could be used as a candidate for gene therapy of non-small cell lung cancer. Methods : After the extraction of total RNA from normal fibroblast cell line and subsequent reverse transcriptase reaction and polymerase chain reaction, the amplified p16 cDNA was subcloned into eukaryotic expression plasmid vector, pRC-CMV. The constructed pRC-CMV-p16 was transfected into the NCI-H441 NSCLC cell line using lipofectin. The changes of G1 cell-cycle related proteins were investigated with Western blot analysis and immunoprecipitation after extraction of proteins from cell lysates and tumor suppressive effect was observed by clonogenic assay. Results : (1) p16(-) NCI-H441 cell line transfected with pRC-CMV-p16 showed the formation of p16 : cdk 4 complex and decreased phosphorylated Rb protein, while control cell line did not. (2) Clonogenic assay demonstrated that the number of colony formation was markedly decreased in p16(-) NCI-H441 cell line transfected with pRC-CMV-p16 than the control cell line. Conclusion : It is confirmed that the expression of p16 protein in p16 absent NSCLC cell line with the gene transfection leads to p16 : cdk4 complex formation, subsequent decrease of phosphorylated pRb protein and ultimately tumor suppressive effects. And also it provides the foundation for the application of p16 gene as a important candidate for the gene therapy of NSCLC.

• Molecules and Cells
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• v.20 no.1
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• pp.43-50
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• 2005

Glutaredoxin (Grx) is a small, heat-stable redox protein acting as a multi-functional glutathione (GSH)-dependent disulfide oxidoreductase. We have cloned the monothiol Grx5 gene from the genomic DNA of the fission yeast Schizosaccharomyces pombe. It has 1,904 bp, with one intron, and encodes a putative protein of 146 amino acids with a molecular mass of 16.5 kDa. Recombinant Grx5 produced functional Grx in S. pombe cells. NO-generating sodium nitroprusside (SNP, 1.0 and 2.0 mM) and potassium chloride (KCl, 0.2 and 0.5 M) increased the synthesis of ${\beta}$-galactosidase from a Grx5-lacZ fusion gene, and transcription of Grx5 was also enhanced by SNP and KCl. Synthesis of ${\beta}$-galactosidase from the Grx5-lacZ fusion was lower in Pap1-negative TP108-3C cells than in wild type KP1 cells, and when Pap1 was overproduced in KP1 cells, the level of ${\beta}$-galactosidase increased. We also found that Pap1 is involved in the induction of Grx5 by SNP and KCl. S. pombe Grx5 may play a crucial role in responses to nitrosative and osmotic stresses.