Background: The MDM2 oncogene, a negative regulator of p53, has a functional polymorphism in the promoter region (SNP309) that is associated with multiple kinds of cancers including non-melanoma skin cancer. SNP309 has been shown to associate with accelerated tumor formation by increasing the affinity of the transcriptional activator Sp1. It remains unknown whether there are other factors involved in the regulation of MDM2 transcription through a trans-regulatory mechanism. Methods: In this study, SNP309 was verified to be associated with overexpression of MDM2 in tumor cells. Bioinformatics predicts that the T to G substitution at SNP309 generates a stronger E2F1 binding site, which was confirmed by ChIP and luciferase assays. Results: E2F1 knockdown downregulates the expression of MDM2, which confirms that E2F1 is a functional upstream regulator. Furthermore, tumor cells with the GG genotype exhibited a higher proliferation rate than TT, correlating with cyclin D1 expression. E2F1 depletion significantly inhibits the proliferation capacity and downregulates cyclin D1 expression, especially in GG genotype skin fibroblasts. Notably, E2F1 siRNA effects could be rescued by cyclin D1 overexpression. Conclusion: Taken together, a novel modulator E2F1 was identified as regulating MDM2 expression dependent on SNP309 and further mediates cyclin D1 expression and tumor cell proliferation. E2F1 might act as an important factor for SNP309 serving as a rate-limiting event in carcinogenesis.
Effects of dietary ${\delta}$-aminolevulinic acid (ALA) supplementation on serum iron status, blood characteristics, egg production and quality were examined in laying hens in an 8-week feeding trail. Two hundred and forty (Hy-line brown, 40-week-old) layers were randomly assigned to four dietary treatments with ten replications (six layers in adjacent three cages). Dietary treatments included: 1) CON (basal diet), 2) ALA1 (CON+ALA 5 ppm), 3) ALA2 (CON+ALA 10 ppm) and 4) ALA3 (CON+ALA 15 ppm). All nutrient levels of diets were formulated to meet or exceed NRC (1994) recommendations for laying hens. During the entire experimental period, differences of serum iron concentration and total iron binding capacity (TIBC) were significantly increased in ALA1 supplemented treatment (quadratic effect, p<0.05). The difference of total protein between 8 and 0 weeks was significantly higher in ALA2 treatment than CON treatment (quadratic effect, p<0.05). No significant effects were observed on hemoglobin, WBC, RBC, lymphocyte and albumin concentrations. Egg production and egg weight were not influenced by the ALA supplementation. Egg yolk index was also significantly higher in ALA3 treatment than CON treatment at the end of 4 and 8 weeks (linear effect, p<0.05). Haugh unit was increased in ALA3 treatment compared to CON and ALA1 treatments at the end of 8 weeks (linear effect, p<0.05). However, egg shell thickness, breaking strength and yolk color unit were not affected by the ALA supplementation. In conclusion, dietary ALA supplementation at a level of 5 ppm can affect iron concentration in serum while higher levels (10 or 15 ppm) have some beneficial influences on blood profiles and egg quality.
Effects of very low level of lead in diet and exposure time on the accumulation and distribution in organs and tissues was investigated with growing rats. 21 days old Sprague-Dawley rats were exposed to lead for 7, 14 and 21 days by feeding of 0.03, 0.42, 0.92 and 1.46mg/kg Pb as Pb-acetate containing diet, respectively. Lead concentrations in blood, liver, kidney and bone exhibit a linear relationship with lead levels in diet. After 7 days of exposure, the greatest dose dependent accumulation of lead was found in kidney and followed in bone. However, after 14 and 21 days, the dose dependent accumlation of lead in bone was about two fold greater than that in kidney. The accumulation of lead in liver and blood was relatively low. As continuous exposure to lead, the concentrations of lead in liver, kidney, blood and intestinal tracts were rather not increased with exposure time. However, bone lead concentration was increased with exposure time by feeding of 0.92 and 1.46mg/kg Pb in diet, but not 0.42mg/kg. The lead concentration in gastrointestinal tracts tends also to increasing with lead levels in diet after 7 and 14 days of exposure. However, by 21 days of exposure the lead concentration revealed relatively constant value regardless of the dietary lead levels. It is concluded that the binding capacity of the lead in blood, liver, kidney and bone seems to be increased with increasing lead levels in diet. The lead concentration in these organs, with the exception of the lead in bone, seems, however, to be standing under steady state regulation by continued exposure with the same dietary lead level. Therefore, by chronic exposure condition with environmental relevant lead level bone might be a principle targe organ for lead and blood lead repesents better the current lead exposure than the lead body burden.
Journal of the Korean Society of Food Science and Nutrition
/
v.26
no.1
/
pp.17-24
/
1997
The effects of steeping on the physicochemical and gelatinization characteristics of glutinous rice flour and its starch were studied. Steeping conditions were 1 day at 25"C,7 days at 2iC and 7 days at 35"C. Crude protein, lipid and ash content were decreased br steeping. It was observed with scanning electron microscopy that endosperm cell wall of glutinous rice flour was diminished by steeping. Although morphology of the glutinous rice starch granules was not affected, the size was decreased by steeping. Density and water binding capacity(WBC) of glutinous rice flour and its starch were changed by steeping. X-ray diffraction pattern of glutinous rice starch was A type and was not affected by steeping. Swelling power of glutinous rice flour and its starch was increased but solubility was decreased by steeping. In Brabender amylographic examination, peak viscosity of untreated glutinous rice flour was very low and increased enormously by steeping resulting in the similar Brabender viscosity pattern to its starch. The gelatinization temperature examined by X-ray diffractometry was lowered by steeping. And the degree of gelatinization under the conclusion temperature increased with increasing of steeping Period and temperature.mperature.
Journal of the Korean Society of Food Science and Nutrition
/
v.22
no.5
/
pp.596-602
/
1993
The Characteristics of Shinsunchalbyeo(Japonica) and Hangangchalbyeo($J{\times}Idica$) starches including physicochemical properties, differential scanning calorimetry(DSC) and enzymatic digestion of lintnerized starches were investigated. Degree of hydrolysis of Hangangchalbyeo starch with 2.2N HCI for 48hr was higher than that of shinsunchalbyeo starch. Absorbance at ${\lambda}_{max}$ 680nm, and ${\lambda}_{max}$ of iodine stained starch decreased upon acid treatment. But water binding capacity, swelling power and solubility considerably increased as hydrolysis progressed. Relative crystallinity of two starches increased with acid treatment, and that of Shinsunchalbyeo starch was higher than that of Hangangchalbyeo starch. DSC data continuously decreased for lintnerization periods, and those of Shinsunchalbyeo starch. DSC data continuously decreased for lintnerization periods, and those of Shinsunchalbyeo starch have higher than those of Hangangchalbyeo starch. The onset temperature of starch by DSC continuously decreased by treatment, but conclusion temperature increased until 24hr and then decreased. The enthalpy for gelatinization decreased for both starches. Degree of hydrolysis of lintnerized Shinsunchalbyeo starch with glucoamylase was slightly higher than that of Hangangchalbyeo starch.
Wide-line 1H NMR intensity and differential scanning calorimetry measurements were carried out on the intrinsically disordered 73-residue full transactivation domain (TAD) of the p53 tumor suppressor protein and two peptides: one a wild type p53 TAD peptide with a helix pre-structuring property, and a mutant peptide with a disabled helix-forming propensity. Measurements were carried out in order to characterize their water and ion binding characteristics. By quantifying the number of hydrate water molecules, we provide a microscopic description for the interactions of water with a wild-type p53 TAD and two p53 TAD peptides. The results provide direct evidence that intrinsically disordered proteins (IDPs) and a less structured peptide not only have a higher hydration capacity than globular proteins, but are also able to bind a larger amount of charged solute ions.
To evaluate iron nutritional status of female college students, fasting blood samples were taken from 76 female students of Kangnung National University. Hemoglobin(Hb), hematocrit(Hct), serum iron(Fe), total iron binding capacity(TIBC) and serum ferritin concentrations were measured and transferrin saturation was calculated. Mean values for Hb, Hct, Fe, TIBC, TS and serum freeitin were 13.64$\pm$1.42g/dl, 40.99$\pm$4.31%, 103.0$\pm$33.3$\mu\textrm{g}$/이, 395.3$\pm$9.07$\mu\textrm{g}$/dl, 26.58$\pm$9.07$\%$and 26.76$\pm$17.5ng/ml, respectively. Prevalence of iron deficiency greatly varied by indices from 6.8% when judged by Hct to 26.0$\%$ by serum ferritin concentration. The Hb concentration was positively correlated with hematocrit (r=0.5402), serum iron(r=0.2819) and transferrin saturation(r=0.2777)(p<0.05). on the other hand, serum ferritin concentration showed significantly negative correlation with TIBC(r=-0.3196). Two-day dietary intake records were collected from subjects to estimate mean daily iron intake and bioavailability of dietary iron. Mean daily intake of iron was 13.15mg and heme iron intake was 0.83mg which was 6.4% of total iron intake. Total absorbable iron calculated by the method of Monsen was 1.27mg and bioavailability of dietary iron was 9.6%. In the light of high prevalence of iron deficiency based of serum ferritin concentration and low bioavailability of iron in the diet, guidelines about diet should be made to increase the content and bioavailability of iron in the diet if female college students.
Journal of the Korean Society for Precision Engineering
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v.23
no.2
s.179
/
pp.199-206
/
2006
Automatic grease lubricator is equipment that provides adequate amount of fresh grease constantly to the shaft and the bearings of machines. It minimizes the friction heat and reduces the friction loss of machines to the least. This research is to develop automatic grease lubricator by gear driven mechanism with controlled operation time. The ultimate design of this equipment is to lubricate an adequate amount of grease by a simple switch clicking according to the advanced set cycle. The backlash of the gear was minimized to increase the output power. To increase the power of gear mechanism, the binding frequency and the thickness of the coil were changed. To control the rotating cycles of the main shaft according to its set numbers, different resistance and chips were used to design the circuit to controls electrical signals with pulse. The body of the lubricator was analyzed by stress analysis with different constructed angle. The stress analysis for differing loading pressures applied to the exterior body of grease lubricator due to the setup angle, was found that the maximum stress was distributed over the outlet part where the grease lubricator suddenly narrowed contracts. Digital mock-up was analyzed and the rapid prototyping(RP) trial products were tested with PCB circuit and grease. The evaluation of the outlet capacity for RP trial products was conducted, because the friction caused by the outlet on the wall surface was an important factor in the operation of the equipment. Finally, the finishing process was applied to decrease the roughness of the surface to a comparable level and was able to test the performance examination for the product.
The optic nerve often suffers regenerative failure after injury, leading to serious visual impairment such as glaucoma. The main inhibitory factors, including Nogo-A, oligodendrocyte myelin glycoprotein, and myelin-associated glycoprotein, exert their inhibitory effects on axonal growth through the same receptor, the Nogo-66 receptor (NgR). Oncomodulin (OM), a calcium-binding protein with a molecular weight of an ~12 kDa, which is secreted from activated macrophages, has been demonstrated to have high and specific affinity for retinal ganglion cells (RGC) and promote greater axonal regeneration than other known polypeptide growth factors. Protamine has been reported to effectively deliver small interference RNA (siRNA) into cells. Accordingly, a fusion protein of OM and truncated protamine (tp) may be used as a vehicle for the delivery of NgR siRNA into RGC for gene therapy. To test this hypothesis, we constructed OM and tp fusion protein (OM/tp) expression vectors. Using the indirect immunofluorescence labeling method, OM/tp fusion proteins were found to have a high affinity for RGC. The gel shift assay showed that the OM/tp fusion proteins retained the capacity to bind to DNA. Using OM/tp fusion proteins as a delivery tool, the siRNA of NgR was effectively transfected into cells and significantly down-regulated NgR expression levels. More importantly, OM/tp-NgR siRNA dramatically promoted axonal growth of RGC compared with the application of OM/tp recombinant protein or NgR siRNA alone in vitro. In addition, OM/tp-NgR siRNA highly elevated intracellular cyclic adenosine monophosphate (cAMP) levels and inhibited activation of the Ras homolog gene family, member A (RhoA). Taken together, our data demonstrated that the recombinant OM/tp fusion proteins retained the functions of both OM and tp, and that OM/tp-NgR siRNA might potentially be used for the treatment of optic nerve injury.
Effect of surface hydrophobicity of soybean peptides on serum cholesterol in rats was investigated. Soybean protein(ISP), casein(CNP), and their peptic hydrolyzates fractionated by acid precipitations (SHT, SH8, SH6, SH4, CHT, CH6, CH5, CH4) were fed to rats and the concentration of serum cholesterol and the fecal steroid excretion were measured. And surface hydrophobicities of the peptide fractions were measured by determining by the ANS flourescence intensity and SDS binding capacity. It was found that the higher the surface hydrophobicity of peptides was, the more the fecal steroids excreted(r=0.801) and the lower the concentration of serum cholesterol became(r=-0.868). However, there was no relationship between SDS surface hydrophobicity and fecal steroids or serum cholesterol. ANS surface hydrophobicity of soybean protein was increased by enzymatic hydrolysis. These results suggest that high surface hydrophobicity of peptides formed during digestion is responsible for the hypocholestrolemic effect of soybean protein through the hydrophobic interaction between the peptides and bile salts in rats.
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