• BMB Reports
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• v.39 no.2
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• pp.216-222
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• 2006

In the present study, we compared six different solubilization buffers and optimized two-dimensional electrophoresis (2-DE) conditions for human lymph node proteins. In addition, we developed a simple protocol for 2-D gel storage. Efficient solubilization was obtained with lysis buffers containing (a) 8M urea, 4% CHAPS (3-[(3-cholamidopropyl) dimethylammonio]-1-propanesulfonate), 40 mM Tris base, 65 mM DTT(dithiothreitol) and 0.2% carrier ampholytes; (b) 5M urea, 2M thiourea, 2% CHAPS, 2% SB 3-10 (N-decyl-N, N-dimethyl-3-ammonio-1-propanesulfonate), 40mM Tris base, 65 mM DTT and 0.2% carrier ampholytes or (c) 7M urea, 2M thiourea, 4% CHAPS, 65 mM DTT and 0.2% carrier ampholytes. The optimal protocol for isoelectric focusing (IEF) was accumulated voltage of 16,500 Vh and 0.6% DTT in the rehydration solution. In the experiments conducted for the sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), best results were obtained with a doubled concentration (50 mM Tris, 384 mM glycine, 0.2% SDS) of the SDS electrophoresis buffer in the cathodic reservoir as compared to the concentration in the anodic reservoir (25 mM Tris, 192 mM glycine, 0.1% SDS). Among the five protocols tested for gel storing, success was attained when the gels were stored in plastic bags with 50% glycerol. This is the first report describing the successful solubilization and 2D-electrophoresis of proteins from human lymph node tissue and a 2-D gel storage protocol for easy gel handling before mass spectrometry (MS) analysis.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1997.04a
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• pp.116-116
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• 1997

Complex formation of practically insoluble dexamethasone dipropionate (DDP) with ${\beta}$-cyclodextrin (${\beta}$-CD), dimethyl-${\beta}$-cyclodextrin (DMCD), trimethyl-${\beta}$-cyclodextrin (TMCD), 2-hydroxypropyl-${\beta}$-cyclodextrin (HPCD) and sulfobutyl ether ${\beta}$-cyclodextrin (SBCD) in water was investigated by solubility method at various temperatures. Water solubility of DDP was found to be 1.78 $\mu\textrm{g}$/$m\ell$ at 37$^{\circ}C$. Propylene glycol (PG)-water cosolvent increased the solubility of DDP, but the solubilization was not sufficient (8.93 $\mu\textrm{g}$/$m\ell$ in 20% PG). The addition of CD markedly increased the solubility of DDP in water, and A$\sub$L/ type phase solubility diagrams were obtained with ${\beta}$-CD, TMCD, HPCD and SBCD, where the apparent stability constants of the soluble complexes at 25$^{\circ}C$ were determined to be 1388, 216, 1054, and 1992 M$\^$-1/, respectively. However, DMCD remarkably increased the solubility of DDP, and showed an A$\sub$P/ type diagram, suggesting that DMCD forms a soluble complex of high order with DDP. The stability constant for the DDP-DMCD complex at 25$^{\circ}C$ was determined to be 19132 M$\^$-1/. The thermodynamic parameters were calculated for the inclusion complex formation in aqueous solution. CD (1${\times}$10$\^$-2/M) remarkably decreased the partition coefficients of DDP between isopropyl myristate/water in the order of TMCD < ${\beta}$-CD < HPCD < SBCD < DMCD, and in squalane/water system in the order of HPCD < TMCD < ${\beta}$-CD < DMCD < DMCD $\leq$ SBCD. This finding represents that, in a o/w type cream, cyclodextrin complexation with DDP may result in high concentration of DDP in aqueous phase. The permeation of DDP through a cellophane membrane was highly suppressed by the addition of CD, and the degree of suppression was different among CDs, indicating that CD may control the skin permeation of DDP. The dissolution rates of solid dispersions with CDs were much faster than those of drugs alone and corresponding physical mixtures. All DDP-CD solid dispersions exceeded the equilibrium solubility. Consequently these results suggest that complex formation of DDP with CDs may provide useful means to markedly enhance the solubility, and CDs are useful in the semi-solid preparations such as creams and gels for topical application.

• Journal of Food Hygiene and Safety
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• v.31 no.6
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• pp.445-450
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• 2016

Thermal-stable soluble proteins (TSSP) in livestock products has been recently reported. Therefore, the development of antibodies and immunoassay using a TSSP is useful because the presence of TSSP can be measured on processed food. In this study, the existence of TSSPs in pork fat and meat was confirmed and their antigenicity was investigated. The extracts from pork fat and meat by heating method were analyzed by SDS-PAGE with 5% stacking and 12% separating gels. The protein profiles from the raw pork fat and meat extracts (major band ranged 25 to 100 kDa) without cooking and heating treatments were significantly different compared to those from cooked and heated pork fat and meat extracts (several major bands > 100 kDa and < 30 kDa). This meant that non thermal-stable soluble proteins ranged from 25 to 100 kDa may be denaturated to insoluble proteins by cooking and heating treatments, and TSSPs were in pork fat and meat at kept their properties. The confirmed TSSPs were used as an immunogen to investigate their antigenicity. Eight mice (5 mice for pork fat and 3 mice for pork meat) were separately immunized with the TSSPs of pork fat and meat, and the anti-sera obtained from the immunized mice showed high titer values. Polyclonal antibodies against each target protein showed the specific reaction to pork fat and meat, individually. These indicated that TSSP could be used as an immunogen to produce antibodies such as monoclonal and polyclonal antibodies. In addition, antibodies specific to TSSP from pork fat and meat may be used as a bio-receptor in immunoassays for the identification of fraudulent adulteration with pork fat and meat in livestock products.

• Korean Journal of Food Science and Technology
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• v.36 no.1
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• pp.38-43
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• 2004

Effects of heat treatment on functional properties of soy protein were examined. Soy protein isolate (SPI) was prepared from Korean soybean varieties, Manli and Taekwang, subjected to heat treatment at $60^{\circ}C$ for 30, 60, 90, and 120 min. pH-solubility results of SPI showed typical U-shape profiles with minimum solubility at pH 4-5 of isoelectric points of soy proteins, longer heat treatments showing slightly higher solubility. Water absorption, emulsifying activity, emulsion stability, and emulsion capacity of SPI increased, while oil absorption decreased, with heating time in Manli variety. Manli and Taekwang showed the highest emulsion capacities after 90-and 60-min heat treatments, respectively. Foam expansion of all SPIs increased with heating time up to 90 min. Texture profile analysis showed heat treatment up to 90 min significantly increased hardness, adhesiveness, springiness, gumminess, and chewiness, whereas significantly decreased cohesiveness of SPI gels (p<0.05).

• Journal of Pharmaceutical Investigation
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• v.35 no.6
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• pp.417-422
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• 2005

This study was aimed to design and formulate the moisture-activated patches containing ofloxacin and lidocaine for antibacterial and local anesthetic action. The solubility of lidocaine at $32^{\circ}C$ in various vehicles decreased in the rank order of PG $759.5{\pm}44.5\;mg/mL$ > PGL > IPM > PEG 300 > PEG 400 > Ethanol > PGMC > DGME > PGML > OA > $Captex^{\circledR}\;300$ > $Captex^{\circledR}\;200$ > water $(4.0{\pm}0.1\;mg/mL)$. Ofloxacin revealed very low solubility, which the highest solubility was obtained from PEG 400 $(18.7{\pm}6.3\;mg/mL)$ among the vehicles used. The addition of lactic acid increased the solubility of ofloxacin dramatically; the solubility at 5% lactic acid was $133.7{\pm}9.7\;mg/mL$. As $2-hydroxypropyl-{\beta}-cyclodextrin$ was added at the concentrations of 40, 80, 120, 160 and 200 mM, the solubilities of lidocaine and ofloxacin were enhanced up to three and two times, respectively, with concentration-dependent pattern. Gel intermediates for filmtype patches were prepared with mucoadhesive polymer, viscosity builders, lidocaine or ofloxacin at pH values from 5 to 7. Gels were cast onto a release liner and dried at room temperature. Dried patch was attached onto an adhesive backing layer, thus forming a patch system. Patches containing a single drug component were characterized by in vitro measurement of drug release rates through a cellulose barrier membrane. The release study was carried out at $37^{\circ}C$ using a Franz-type cell. Receptor solutions were isotonic phosphate buffers (pH 7.4). Samples $(100\;{\mu}L)$ were taken over 24 hours and quantitated by a verified HPLC method. The releases from all tested were proportional to the square root of time. The release rates were 0.9, 157.3 and $281.7\;{\mu}g/cm^{2}/min^{1/2}$ for the lidocaine patches and 19.8,37.2 and $50.7\;{\mu}g/cm^{2}/min^{1/2}$ for the ofloxacin patches at the concentrations of 0.3, 0.5 and 1 %, respectively. The release rates were dose dependent in both drug patches $(R^{2}\;=\;0.9077\;for\;lidocaine;\;R^{2}\;=\;0.9949\;for\;ofloxacin)$ and those were also thickness-dependent $(R^{2}\;=\;0.9246\;for\;lidocaine;\;R^{2}\;=\;0.9512\;for\;ofloxacin)$.

• Journal of Animal Science and Technology
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• v.44 no.5
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• pp.633-642
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• 2002

Enteric colibacillosis has economically become an important disease of young animals as a result of increasing intensification of farrowing management. The objective of this experiment is to isolate fimbrial antigen from enterotoxigenic E. coli F41, to develop specific polyclonal IgY which can effectively neutralize or reduce the proliferation of pathogens in feed or living animal system, and to apply IgY technologies to animal industry. The results obtained were as follows: The molecular weight of the purified F41 antigen was 29,500 dalton on sodium dodecyl sulfate-polyacrylamide gels. Fimbrial antigen was confirmed by the western blot method. It was observed that after immunization the level of serum antibody titer of laying hen was shown in two weeks and gradually increased. The antibody titer in egg yolk appeared two weeks after it was shown in serum antibody. The titers of egg yolk antibody were gradually increased to the maximum level of 320,000 (antigen 50${\mu}g$/$m\ell$), 450,000 (antigen 200${\mu}g$/$m\ell$), and 320,000 (antigen 600${\mu}g$/$m\ell$). According to the results of specificity test by ELISA, the anti-F41 antibodies from chicken serum and egg yolk reacted only with ETEC F41 antigen. There was no cross reaction with other ETEC strains (K88, K99, and 987P). In vitro condition, as a result of antigen binding ability of yolk antibodies, bacterial concentration was rapidly decreased to $10^5$ CFU/$m\ell$ from $10^9$ CFU/$m\ell$ when 2${\sim}$4 mg/$m\ell$ of freeze dried WSF (water soluble fraction) was added.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.6
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• pp.2631-2634
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• 2014

Background: p53 gene is a well-known tumor suppressor gene that has several polymorphisms in both its exons and introns. It has been suggested that intron 3 16 bp duplication polymorphism may affect the gene function resulting in reduction or suppression of p53 anti tumor activity. In most case control studies a duplicated allele has been noticeably more frequent in cases rather than controls but there are also conflicting results. The aim of this study was to assess the association of intron 3 16 bp duplication polymorphism of p53 with breast cancer risk among Iranian-Azeri population. We also analyzed the clinicopathological information of patients as an epidemiological description of breast cancer in the north-west of Iran. Materials and Methods: This case-control study was performed on 221 breast cancer patients and 170 controls. Genomic DNA was extracted from peripheral blood samples and tumor tissues. p53 PIN3 genotype was determined using electrophoresis of PCR products on 8% non-denaturing polyacrylamide gels and silver staining. Results: In the control and case groups, respectively, 62.9% and 61.1% had no 16 bp insertion (A1A1 genotype), 7.1% and 7.7% had insertion in both p53 alleles (A2A2) and 30% and 31.2% were heterozygous (A1A2). There was no significant difference between genotype frequencies as well as allelic frequencies in two case and control groups. Conclusions: According to the result of the present study, the intron 3 16 bp duplication polymorphism of p53 could not be assessed as a marker of risk factor for predisposition to breast cancer in Azeri population. However, a high frequency of A2 allele (22.1%) in our population suggested that intron 3 16 bp duplication polymorphism may be a valuable marker for study in other cancers with well designed large groups.

• Applied Biological Chemistry
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• v.26 no.4
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• pp.211-216
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• 1983

The physicochemical properties of starches isolated from three kinds of waxy ricer were investigated. The average diammeter of Orchal starch granule was 3.6u, and those of Hangang and Baegoon starch granules were 4.8u and 5.6u, respectively. X-ray diffraction patterns of all samples were A types. Optical transmittance of starch suspensions was increased rapidly $60{\sim}62^{\circ}C$, $50{\sim}55^{\circ}C$ and $50{\sim}60^{\circ}C$ in Olchal, Hangang and Baegoon, respectively, and all of them exhibited maximum transmittance $75{\sim}80^{\circ}C$ in range. Initial gelationization temperature by means of amylogram pattern was $60{\sim}61^{\circ}C$, then, Olchal starch had a little higher temperature than others. Raising power of them was $260\sim220$, and then, Olchal starch has a little higher raising power. But, water binding capacity of Baegoon and Hangang were somewhat higher than that of Olchal. Blue values of Olchal and Hangang were similar with 0.13, and that of Baegoon was 0. 14. Alkari numbers of Olchal, Hangang and Baegoon were 4.2, 4.9 and 5.1. The degree of retrogradation of Hangang and Baegoon starch gels were somewhat higher than that of Olchal starch gel.

• The Korean Journal of Physiology and Pharmacology
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• v.8 no.4
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• pp.207-211
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• 2004

The purpose of the present study was to evaluate the expression of cardiac marker protein in rabbit cardiac tissue that was exposed to ischemic preconditioning (IPC), or ischemiareperfusion injury (IR) using two-dimensional gel electrophoresis (2DE) and matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS). We compared 2DE gels of control (uninjured) cardiac tissue with those of IPC and IR cardiac tissue. Expression of one protein was detected in IR heart tissue, however the protein was not detected in the samples of control and IPC tissue. To further characterize the detected protein molecule, the protein in the 2D gel was isolated and subjected to trypsin digestion, followed by MALDI-MS. The protein was identified as myoglobin, which was confirmed also by Western blot analysis. These results are consistent with previous studies of cardiac markers in ischemic hearts, indicating myoglobin as a suitable marker of myocardial injury. In addition, the present use of multiple techniques indicates that proteomic analysis is an appropriate means to identify cardiac markers in studies of IPC and IR.

• The Journal of Korean Society for Radiation Therapy
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• v.19 no.1
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• pp.7-17
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• 2007

Purpose: As increasing complexity of modern radiotherapy technique, more developing dosimetry is required. Polymer gel dosimeters offer a wide range of potential applications with high resolution and assured quality in the thee-dimensional verification of complex dose distribution such as intensity-modulated radiotherapy (IMRT). The purpose of this study is to find the most sensitive and suitable gel as a dosimeter by varying its composition ratio and its condition such as temperature during manufacturing. Materials and Methods: Each polymer gel with various ratio of composition was irradiated with the same amount of photon beam accordingly. Various polymer gels were analyzed and compared using a dedicated software written in visual C++ which converts TE images to R2 map images. Their sensitivities to the photon beam depending on their composition ratio were investigated. Results: There is no dependence on beam energy nor dose rate, and calibration curve is linear. Conclusion: The polymer gel dosimeter developed by using anti-oxidant in this study proved to be suitable for dosimetry.