• Korean Journal of Veterinary Research
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• v.36 no.3
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• pp.577-598
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• 1996

This study was conducted to identify the effects of caffeine or combinations of caffeine and iron or vitamin E on the lipid and protein components or blood chemistry levels of the serum as well as the total homogenate, mitochondrial and microsomal fraction of the rat(Sprague-Dawley, female) liver. Chronic test were conducted to determine those effects. The chronic test was conducted by dividing rats into 5 groups according to the type of drugs and dosages administrated as follows; the control(group A), and group B was given 25mg/kg caffeine orally once daily for 30 days, group C was given 50mg/kg caffeine orally once daily for 30 days, group D was given 25mg/kg caffeine and orally ferric chloride once daily for 30 days and group E was given 25mg/kg caffeine and 25mg/kg vitamin E once daily for 30 days. The concentrations of glucose, urea nitrogen, uric acid, creatinine, total protein, albumin, A/G ratio, triglyceride, total cholesterol, HDL-cholesterol, free fatty acid, phospholipid as well as the activities of alanine aminotransferase(ALT), aspartate aminotransferase(AST) and alkaline phosphatase(ALP) were measured in the serum of each experimental groups. The concentrations of the carbonyl group and malondiaidehyde(MDA) and the patterns of the SDS-PAGE(Sodium Dodecyl Sulfate - Polyacrylamide Gel Electrophoresis) and fatty acid compositions in free fatty acids and phospholipids were analyzed to determine the oxidative damages and metabolic changes on the lipid and protein components in the serum, and total homogenate, mitochondrial and microsomal fractions of the rat liver. The results obtained from this study were summarized as follows; 1. Body weights of groups B, C, D and E were significantly decreased(p < 0.01) in comparison with that of the control in the chronic test. 2. The concentrations of serum glucose in groups B(124.5mg/dl), C(130.1mg/dl), D(122.1mg/dl), E(119.3mg/dl) were significantly higher(p < 0.01) in comparison to that of the control(101.5mg/dl). But, there were no significant differences in the concentrations of urea nitrogen, uric acid, creatinine, total protein, albumin and A/G ratio in comparison to that of the control. 3. The concentrations of total cholesterol and HDL-cholesterol in serum of groups B(69.6, 53.4mg/dl), C(73.0, 56.3mg/dl), D(68.9, 51.1mg/dl) and E(68.2, 51.3mg/dl) were significantly higher(p < 0.01) in comparison to that of the control(52.6, 38.8mg/dl). On the other hand, the concentrations of triglyceride in serum of groups B(45.0mg/dl), C(40.4mg/dl), D(33.8mg/dl) and E(47.2mg/dl) were significantly lower(p < 0.01) in comparison to that of the control(66.2mg/dl). There were no significant differences in the activities of ALT, AST and ALP in comparison to that of the control. 4. The concentrations of free fatty acid and phospholipid in serum of groups B(45.7, 154.4mg/dl), C(50.0, 167.2mg/dl), D(52.5, 148.4mg/dl) and E(41.1, 159.2mg/dl) were higher(p < 0.01) in comparison to that of the control(35.2, 125.3mg/dl). And the concentrations of the carbonyl group and malondialdehyde in serum of group D(1.82, 0.52nM/mg protein) were significantly higher(p < 0.01) in comparison to the control(1.53nM/mg protein). 5. The concentrations of carbonyl group in total homogenate, mitochondrial and microsomal fraction of group D(1.45, 0.94, 1.67nM/mg protein) were significantly higher (p < 0.01) in comparison to the control(1.16, 0.66, 1.27nM/mg protein). And the concentrations of malondialdehyde in the total homogenate, mitochondrial and microsomal fraction of group D(6.70, 6.10, 1.36nM/mg protein) were significantly higher(p < 0.01) in comparison to the control(5.17, 3.64, 0.68nM/mg protein). 6. As the analytical results of the fatty acid compositions of free fatty acid in serum, the proportions of stearic acid and arachidonic acid of groups B(16.52, 12.62%), C(17.52, 15.18%), D(19.73, 13.47%) and E(17.62, 13.28%) were significantly higher(p < 0.01) in comparison to the control(14.75, 7.88%), but the proportions of oleic acid and linoleic acid of groups B(12.97, 32.59%), C(10.88, 31.23%), D(12.37, 30.66%) and E(11.95, 32.41%) were significantly lower(p < 0.01) in comparison to the control(16.44, 35.12%). Otherwise, as the results of the fatty acid compositions of phospholipid in serum, the proportions of stearic acid and arachidonic acid of groups B(39.37, 16.39%), C(40.63, 17.83%), D(42.73, 15.39%) and E(39.16, 15.70%) were significantly higher(p < 0.01) in comparison to the control(37.74, 14.24%), but the proportions of oleic acid and linoleic acid of groups B(4.03, 14.38%), C(3.54, 12.38%), D(4.52, 11.68%) and E(4.29, 13.64%) were significantly lower(p < 0.01) in comparison to the control(5.53, 16.14%). 7. As the analytical results of the fatty acid compositions of free fatty acid in total homogenate, mitochondrial and microsomal fraction of liver, the proportions of oleic acid of groups B(7.8**, 8.73**, 6.88%) and C(6.89**, 7.75**, 6.58%) were lower(**:p < 0.01) in comparison to the control(8.67, 10.08, 7.81%), but the proportions of arachidonic acid of group C(22.62, 19.79, 23.71%) were significantly higher(p < 0.01) in comparison to the control(20.93, 18.47, 22.24%). And the proportions of palmitic acid of group D(25.95**, 26.16, 26.34**%) were significantly higher(**:p < 0.01) in comparison to the control(24.43, 25.42, 23.34%). In addition, the proportions of linoleic acid of group D(23.43, 25.02, 23.95%) were also significantly higher(p < 0.01) in comparison to the control(22.17, 23.75, 21.26%). The proportions of stearic acid of group D(19.87, 19.76**%) in mitochondrial and microsomal fraction were lower(**:p < 0.01) in comparison to the control(21.01, 24.18%), and the proportions of stearic acid of group E(16.71*, 19.65**%) in mitochondrial and microsomal fraction were significantly lower(**:p < 0.01, *:p < 0.05) in comparison to the control(21.01, 24.18%), and the proportions of linoleic acid of group E(25.04, 29.20, 26.48%) in total homogenate, mitochondria and microsome were significantly higher(p < 0.01) in comparison to the control(22.17, 23.75, 21.26%). 8. As the results of the fatty acid compositions of phospholipid in total homogenate, mitochondrial and microsomal fraction of liver, the proportions of palmitic acid of group D(17.58**, 18.78*, 18.23%**) were significantly higher(**:p < 0.01, *:p < 0.05) in comparison to the control(16.28, 17.22, 16.38%), and the proportions of stearic acid of group D(36.41, 37.23, 39.53%) were also significantly higher(p < 0.01) in comparison to the control(34.18, 34.16, 36.04%). But the proportions of oleic acid(3.41*, 3.11**, 3.12**%) and linoleic acid (18.03**, 15.79**, 14.74**%) of group D were significantly lower(**:p < 0.01, *:p < 0.05) in comparison to the control(oleic : 3.63, 3.72, 3.79%, linoleic : 20.03, 18.71, 18.48%). 9. In order to determine the oxidative damages to the protein in serum, mitochondrial and microsomal fraction of the rat liver, the patterns of the SDS-PAGE were identified, but the results of SDS-PAGE were not significantly different between the control and experimental groups.

• Journal of Korean Neurosurgical Society
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• v.29 no.4
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• pp.471-476
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• 2000

Objective : p16/INK4a, a kind of tumor suppressor genes, encodes a specific inhibitor of the cyclin D-dependent kinases CDK4 and CDK6. This prevents the association of CDK4 with cyclin D1, and subsequently inhibits phosphorylation of retinoblastoma tumor suppressor protein(pRb), thus preventing exit from the G1 phase. According to previous reports, over 50% of glioma tissue and 80% of glioma cell lines have been demonstrated inactivation of p16/INK4a gene. The purpose of this study was to determine whether recombinant adenovirus-p16 virus is a suitable candidate for gene replacement therapy in cases of glioma. Methods : Three human glioma cell lines(U251MG, U87MG and U373MG) that express mutant p16 protein were used. Replication-deficient adenovirus was utilized as an expression vector to transfer exogenous p16 cDNA into the cells ; control cells were infected with the Ad-${\beta}$-gal expressing ${\beta}$-galactosidase. To monitor gene transfer and the expression of exogenous genes, we used Western Blotting analysis. Flow cytometry studies of cellular DNA content were performed to determine the cell cycle phenotype of the glioma cells before and after treatment. Results : We showed here that restoration of p16/INK4a expression in p16 negative U87MG, U251MG and partially deleted U373MG by Ad-CMV-p16 induced growth suppression in vitro. Flow cytometric study revealed that Ad-CMV-p16 infected U87MG cells were arrested during the G0-G1 phase of the cell cycle. Expression of p16 transferred by Ad-CMV-p16 in glioma cells was highly efficient and maintained for more than seven days. Conclusions : Our results suggest that Ad-CMV-p16 gene therapy strategy is potentially useful and warrants further clinical investigation for the treatment of gliomas.

• Asian-Australasian Journal of Animal Sciences
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• v.9 no.6
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• pp.727-735
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• 1996

The effects of four factors-two dietary protein levels (12 and 16%), feeding methods(ad libitum and restricted), strains (A and B), and age of hens-on egg quality and laying performance were examined. Use of the 12% dietary protein level resulted in significantly lower (p < 0.05) hen-day egg production, higher feed intake to about 85% of the ad libitum intake decreased mean hen-day production. Significant feeding method-by-age interactions ($F{\times}A$) were obtained for all parameters. Restricted feeding had favorable effects on shell strength and Haugh unit values after 28 and 16 weeks of egg production, respectively. There was significant difference in shell strength between the two strain. Examination of the three-factor interaction among protein levels, feeding method and strain ($P{\times}F{\times}S$) disclosed that the favorable effect of the A strain on shell strength was significant only with the 16% protein-restricted fed group. The other treatment group did show a trend for greater shell strength of the A strain. Mean values for all the parameters examined changed significantly (p < 0.01) with the age of hens. Feed intake per dozen eggs tended to increase, with some fluctuation, as the hens aged. There was a linear decrease in Haugh unit scores and shell strength. The effect of restricted feeding on either shell strength or Haugh unit scores were favorable for the aged chickens.

• Applied Biological Chemistry
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• v.19 no.1
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• pp.31-35
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• 1976

Fifteen naked barley cultivars including radiation breeding lines from three places were analized for crude protein, carbohydrates, P, K, Ca, Mg and tested protein by dye binding method and biuret method. Their content and simple correlation analyses among them were as follows. 1. Protein content was 7.67 for average (max. 10.3 in Baegdong, min. 6.0 in Bangju) that was lower than in milled barley and had significant (at p=0.01) correlation with dye binding capacity (r=0.769) and biuret absorbance (r=0.616). 2. Protein content also had significant correlation with $P_2O_5$(r=0.607, p=0.01) and with MgO(r=0.498, p=0.05). 3. There was great difference in protein content among radiation breeding lines(max. 8.40, min. 6.75%). 4. Naked barley appeared to be lower in carbohydrate content but higher in crude ash to compare with milled barley. 5. There was significant correlation(r=0.560, p=0.01) between Ca and K, indicating competition in uptake or translocation to grain. 6. Carbohydrate content showed the highest negative correlation with protein content but it was not significant. 7. The low protein variety (Bangju) showed higher yield than the high protein one (Baegdong) both with (16%) and without (48%) fertilizers.

• Asian Pacific Journal of Cancer Prevention
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• v.14 no.5
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• pp.2711-2715
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• 2013

Background: Cancer of the gallbladder is a relatively rare neoplasm with a poor prognosis. The exact mechanisms of its genesis are not known and very little information is available on molecular events leading to labeling this as an orphan cancer. Materials and Methods: In this prospective case control study we evaluated the expression of p16, pRb and cyclin D1 by immunohistochemistry to study the G1-S cell-cycle check point and its possible role in gallbladder carcinogenesis. A total of 25 patients with gallbladder carcinoma (group I), 25 with cholelithiasis (group II) and 10 normal controls. were enrolled Results: Cyclin D1 expression was seen in 10 (40%) patients each with carcinoma and cholelithiasis while only in 2 (20%) of the normal gallbladders but differences were not statistically significant (p value=0.488). p16 was expressed in 12% patients of carcinoma of the gallbladder and 28% of cholelithiasis, however this difference was not statistically significant (p value=0.095). Retinoblastoma protein was found to be expressed in 50% of normal gallbladders and 6 (24%) of carcinoma and 8 (32%) of gallstones. The present study failed to demonstrate any conclusive role of cyclin D1/RB/ p16 pathway in carcinoma of the gallbladder. Conclusions: The positive relation observed between tumor metastasis and cyclinD1 expression and p16 with nodal metastasis suggested that higher cyclin D1/p16 expression may act as a predictive biomarker for aggressive behavior of gallbladder malignancies.

• Asian-Australasian Journal of Animal Sciences
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• v.25 no.7
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• pp.971-979
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• 2012

Four, lactating dairy crossbreds ($50%{\times}50%$ Holstein Friesian${\times}$Native Zebu cattle) were randomly assigned according to a $2{\times}2$ factorial arrangement (two protein levels and two levels of mangosteen peel pellets (Mago-pel)) in a $4{\times}4$ Latin square design to receive four dietary treatments. All cows received concentrate at a proportion of 1 kg concentrate per 2 kg of milk yield, and urea-treated 5% rice straw (UTRS) was given ad libitum. It was found that total dry matter intakes, nutrient digestibility, ruminal pH and $NH_3$-N concentrations were not affected (p>0.05) by treatments. Concentrations of ruminal pH and $NH_3$-N were not affected by dietary treatments although the concentration of BUN varied significantly (p<0.05) between protein levels (p<0.05). The populations of rumen bacteria and fungal zoospores did not differ among treatments (p>0.05); however, the population of protozoa was decreased (p<0.05) when cows received Mago-pel supplementation. The composition of the population of bacteria, identified by real-time PCR technique, including total bacteria, methanogens, Fibrobacter succinogenes and Ruminococcus albus was similar (p>0.05) among dietary treatments (p>0.05); however, copy numbers of Ruminococcus flavefaciens was increased when protein level increased (p<0.05). Microbial protein synthesis, in terms of both quantity and efficiency, was enriched by Mago-pel supplementation. Milk yield was greatest in cows fed UTRS based diets with concentrate containing protein at 16% CP with Mago-pel, but were lowest without Mago-pel (p<0.05). In addition, protein level and supplementation of Mago-pel did not affect (p>0.05) milk composition except solids-not-fat which was higher in cows fed the diet with 19% CP. Therefore, feeding a concentrate containing 16% CP together with 300 g/hd/d Mago-pel supplementation results in changes in rumen fermentation and microbial population and improvements in milk production in lactating dairy crossbreds fed on UTRS.

• Asian Pacific Journal of Cancer Prevention
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• v.16 no.9
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• pp.3723-3727
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• 2015

The p53 tumor suppressor protein is a principal mediator of growth arrest, senescence, and apoptosis in response to a broad array of cellular damage. p53 is a substrate for the ubiquitin-proteasome system, however, the ubiquitin-conjugating enzymes (E2s) involved in p53 ubiquitination have not been well studied. UBE2Q1 is a novel E2 ubiquitin conjugating enzyme gene. Here, we investigated the effect of UBE2Q1 overexpression on the level of p53 in the MDA-MB-468 breast cancer cell line as well as the interaction between UBE2Q1 and p53. By using a lipofection method, the p53 mutated breast cancer cell line, MDA-MB-468, was transfected with the vector pCMV6-AN-GFP, containing UBE2Q1 ORF. Western blot analysis was employed to verify the overexpression of UBE2Q1 in MDA-MB-468 cells and to evaluate the expression level of p53 before and after cell transfection. Immunoprecipitation and GST pull-down protocols were used to investigate the binding of UBE2Q1 to p53. We established MDA-MB-468 cells that transiently expressed a GFP fusion proteins containing UBE2Q1 (GFP-UBE2Q1). Western blot analysis revealed that levels of p53 were markedly lower in UBE2Q1 transfected MDA-MB-468 cells as compared with control MDA-MB-468 cells. Both in vivo and in vitro data showed that UBE2Q1 co-precipitated with p53 protein. Our data for the first time showed that overexpression of UBE2Q1can lead to the repression of p53 in MDA-MB-468 cells. This repression of p53 may be due to its UBE2Q1 mediated ubiquitination and subsequent proteasome degradation, a process that may involve direct interaction of UBE2Q1with p53.

• Genomics & Informatics
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• v.12 no.2
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• pp.64-70
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• 2014

Human papillomavirus (HPV) infection is the leading cause of cancer mortality among women worldwide. The life-threatening infection caused by HPV demands the need for designing anticancerous drugs. In the recent years, different compounds from natural origins, such as carrageenan, curcumin, epigallocatechin gallate, indole-3-carbinol, jaceosidin, and withaferin, have been used as a hopeful source of anticancer therapy. These compounds have been shown to suppress HPV infection by different researchers. In the present study, we explored these natural inhibitors against E6 oncoprotein of high-risk HPV-16, which is known to inactivate the p53 tumor suppressor protein. A robust homology model of HPV-16 E6 was built to anticipate the interaction mechanism of E6 oncoprotein with natural inhibitory molecules using a structure-based drug designing approach. Docking analysis showed the interaction of these natural compounds with the p53-binding site of E6 protein residues 113-122 (CQKPLCPEEK) and helped the restoration of p53 functioning. Docking analysis, besides helping in silico validation of natural compounds, also helps understand molecular mechanisms of protein-ligand interactions.

• Environmental Mutagens and Carcinogens
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• v.25 no.3
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• pp.87-96
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• 2005

[ $p16^{INK4a}$ ] is a tumor suppressor that belongs to the INK4 family of the cyclin D-dependent kinases (cdk) inhibitors. It plays regulatory roles in cell proliferation and in tumorigenesis by interacting with Rb signaling. Abnormally elevated $p16^{INK4a}$ protein expression causes cell cycle arrest (G1/S transition) and loss of cyclin-cdk activity. In many cancers, $p16^{INK4a}$ is altered by mutation, deletion, and promoter methylation. This review summarizes the function of p16 as an important regulator of cancer pathobiology and a promising target fer developing cancer therapeutic and chemopreventive agents.

• Journal of the Korean Magnetic Resonance Society
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• v.16 no.1
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• pp.22-33
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• 2012

Tid1, belonging to the Hsp40/DnaJ family of proteins, functions as a cochaperone of cytosolic and mitochondrial Hsp70 proteins. In particular, the N-terminal J-domain of Tid1 (Tid1-JD) constitutes the major binding sites for proteinprotein interactions with client proteins, including p53, as well as its partner chaperone, Hsp70. In the present study, soluble, recombinant protein of Tid1-JD could be obtained by using the pCold vector system, and backbone NMR assignments were completed using the isotope $[^{13}C/^{15}N]$-enriched protein. Far-UV CD result implied that Tid1-JD is an ${\alpha}$-helical protein and the secondary structure determined using chemical shift data sets indentified four ${\alpha}$-helices with a loop region containing the HPD (conserved tripeptide of His, Pro and Asp) motif. Additionally, NMR spectra under different conditions implied that the HPD motif, which is a critical region for protein-protein interactions of Tid1-JD, would possess dynamic properties.