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
/
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.
Purpose : The purpose of this study was to investigate the applicability of poly-L-guluronic alginate (PGA) gel in vascular embolization with angiography simulation. Materials and Methods : To prepare a gel-forming PGA from no guluronate-rich Laminaria japonica, a new acid hydrolysis method was employed with a lower HCL concentration (0.03 M) and a shorter treatment time (5 min). The obtained PGAs were selected based on gel stability and viscosity. Glass aneurysm model was used to simulate gel embolization in vitro. Then, finally, the PGA was used to embolize the renal vascular system by using a rabbit model and angiography. Results : Glass aneurysm model was made to simulate gel embolization procedure. PGA solution was injected from pump through 2-way catheter. Subsequent injection of $CaCl_2$ successfully formed gels inside aneurysm model that conforming to its inner contour. In rabbit model, first, renal artery and aorta leading to the right kidney were ligated to block blood flow, then conventional contrast agent was injected through aorta to check the arterial patency to the left kidney. In sequential artery injection method, PGA and $CaCl_2$ were injected through renal artery sequentially via a single catheter. Re-injection of contrast agent after removing ligated aorta showed blood flow to the right kidney but no flow in the left kidney. This result demonstrated a complete blocking of blood flow due to gel formation in vascular bed of the left kidney. Conclusion : Instillation of calcium alginate into aneurysm model and arterial system in vivo produced an embolization that better fills and conforms to the contour of aneurysms or blocking vascular bed completely. Therefore, PGA was effective endovascular occlusion materials and provide an efficiency of vascular angiography.
The effect of alachlor treatment on protein synthesis was studied. Protein synthesis was inhibited by $1{\times}10^{-4}$ M and $1{\times}10^{-3}$M of alachlor 5.8% and 86.5%, respectively, while did not occur blow $1{\times}10^{-5}$M alachlor. Soluble protein of alachlor treated oat root tips was examined by polyacrylamide gel electrophoresis. The proteins extracted from oat root tips showed that they were made up of subunits blow 100 kd polypeptides by SDS-PAGE. As compared to control, high molecular proteins(above 47 kd) were inhibited of oat root treated with alachlor, while low molecular proteins(below 23 kd) were increased. Two-D gels showed that alachlor caused decrease(1-6 spots) or increase(7-10 spots) in number of polypeptides on silver staining. The intensity of some polypeptides of soluble proteins (molecular mass of 83 kd : 1, 2 spots, 70 kd : 3, 4 spots, and 47.5 kd : 5, 6 spots) decreased in alachlor treatment, whereas the intensity of other peptide bands (20 kd : 7 spot and 16 kd : 8, 9, 10 spots) increased. Oat root tip proteins present in the neutral zone are masked by diffusing of major proteins, but proteins in acid zone are resolved minor proteins.
Background: Hyaluronic acid (HA) has been applied as a primary biomaterial for temporary soft tissue augmentation and as a carrier for cells and the delivery of growth factors to promote tissue regeneration. Although HA derivatives are the most versatile soft tissue fillers on the market, they are resorbed early, within 3 to 12 months. To overcome their short duration, they can be combined with cells or growth factors. The purpose of this study was to investigate the stimulating effects of human fibroblasts and basic fibroblast growth factors (bFGF) on collagen synthesis during soft tissue augmentation by HA hydrogels and to compare these with the effects of a commercial HA derivative (Restylane®). Methods: The hydrogel group included four conditions. The first condition consisted of hydrogel (H) alone as a negative control, and the other three conditions were bFGF-containing hydrogel (HB), human fibroblast-containing hydrogel (HF), and human fibroblast/bFGF-containing hydrogel (HBF). In the Restylane® group (HGF), the hydrogel was replaced with Restylane® (R, RB, RF, RBF). The gels were implanted subdermally into the back of each nude mouse at four separate sites. Twelve nude mice were used for the hydrogel (n = 6) and Restylane® groups (n = 6). The specimens were harvested 8 weeks after implantation and assessed histomorphometrically, and collagen synthesis was evaluated by RT-PCR. Results: The hydrogel group showed good biocompatibility with the surrounding tissues and stimulated the formation of a fibrous matrix. HBF and HF showed significantly higher soft tissue synthesis compared to H (p < 0.05), and human collagen type I was well expressed in HB, HF, and HBF; HBF showed the strongest expression. The Restylane® filler was surrounded by a fibrous capsule without any soft tissue infiltration from the neighboring tissue, and collagen synthesis within the Restylane® filler could not be observed, even though no inflammatory reactions were observed. Conclusion: This study revealed that HA-based hydrogel alone or hydrogel combined with fibroblasts and/or bFGF can be effectively used for soft tissue augmentation.
Adherence of Candida albicans(C. albicans) to the surface of a denture is believed to be an initial and essential step in the formation of denture-induced stematitis. Previous studies have provided enormous infomation on the relationship between composition of palatine gland/parotid saliva and upper denture stomatitis. Relatively little information is available on the correlation between lower denture stomatitis and sublingual-submandibular ( SLSM ) saliva. The plaque samples were collected from the two sites($100mm^2$) on the inner surface of lower partial denture corresponding to the stematitis and healthy region of the lower partial dentures of 12 denture stomatitis patients and 6 nor-mal persons who wore lower partial dentures. The samples were plated to isolate C. albicans on a selective Saboraud's dextrose agar plate and the isolates were identified by germ tube test and gram staining. The subjects were divided into group I (stomatitis with C. albican), group II (lesion without C. albicans), group III (no lesion but C. albicans), and group IV (normal and healthy denture wearer). Individual SLSM saliva($20{\mu}g$ of protein) was analyzed by SDS-PAGE (SDS -poly-acrylamide gel electrophoresis) with Coomassie brilliant blue and PAS(Periodic Acid Schinff) stain-ing. The salivary proteins separated in the polyacryamide gels were subjected to immunoblot anaysis using anti-lactoferrin, anti-sIgA, and anti-secretory component of sIgA. In this study using custom made acrylic denture resin beads(5mm in diameter) coated with stimulated individual SLSM saliva, the binding ability of individual C. albicans strains to the beads was observed. Levels of C, albicans adhered to the acrylic resin beads were determined by measuring the optical density of the bound C. albicans to the beads at 580nm. The results showed that a higher number of C. albicans was observed in the lesion site than healthy site. The saliva of group I contained more high molecular weight glycoprotein(mucin, MGI) as compared to group II, III and IV. And lactoferrin and sIgA affected to the binding ability of C. albicans to acylic resin beads. Binding ability of individual C. albicans to the acrylic resin coated with respective individual saliva was found to be greater in group I than the other 3 groups. And when bound cells of C. albicans isolated from individual subject #2 to the saliva coated beads were used binding ability of subject #2 saliva coated beads was founed to be greater than the other sutjects. These results suggested that denture induced stomatitis is related to individual patient's salivary protein composition, especially MG-1. Future studies will be directed toward saliva exam-ination of patients who have general disease and analysis of pellicles formed on prosthesis with respect to oral disease.
Purpose : Fragile X syndrome (FXS) is the most common heritable cause of cognitive impairment. FXS is caused by hyperexpansion and hypermethylation of a polymorphic CGG trinucleotide repeat in the 5' untranslated region of the fragile X mental retadation-1(FMR1) gene. Combination of Southern blotting and simple polymerase chain reaction(PCR) amplification of the FMR1 repeat region is commonly used for diagnosis in females. To give a definite diagnosis in a female child suspected of having FXS, we carried out the molecular diagnostic test for FXS using the recently developed Abbott Molecular Fragile X PCR Kit. Methods : The PCR amplification of the FMR1 repeat region was performed using the Abbott Mdecular Fragile X PCR Kit. The amplified products were analyzed by size-separate analysis on 1.5% agarose gels and by DNA fragment analysis using Gene scan. Results : Agarose gel and Gene scan analyses of PCR products of the FMR1 repeat region showed that the patient had two heterozygous alleles with a normal 30 repeats and full mutation of >200 repeats whereas her mother had two heterozygous alleles with the normal 30 repeats and premutation of 108 repeats, suggesting that the premutation of 108 repeats in her mother may have led to the full mutation of >200 repeats in the patient. Conclusion : We diagnosed FXS in a female patient using a simplified molecular diagnostic test. This commercially available diagnostic test for FXS, based on PCR, may be a suitable alternative or complement method to Southern blot analysis and PCR analysis and/or methylation specific(MS)-PCR analysis for the molecular diagnosis of FXS in both males and females.
The physicochemical properties and gel-forming properties of corn & red bean crude starches were investigated. The results were as follows: 1. The shape of corn crude starch granule was polygonal and the mean value of minor axis and major axis were $11.5\mu\textrm{m}$ and $14.9\mu\textrm{m}$, respectively. In the meantime, the shape of red bean crude starch granule was oval and the mean value of minor axis and major axis were $22.3\mu\textrm{m}$ and $31.4\mu\textrm{m}$. 2. Amylose content of corn and red bean refined starch were 16.52 and 43.61% respectively. 3. Blue value of corn and red bean crude starch were 0.099 and 0.842, respectively. 4. Amylose of corn had molecular weight of 107,000 and degree of polymerization of 660. Amylopectin had degree of branching of 6.9 per 100 glucose units and glucose units of 14.6 persegment of amylopectin. Amylose of red bean had molecular weight of 118,000 and amylopectin had degree of branching of 5.2. 5. Water binding capacities of corn and red bean starch were 238.5 and 284.8. 6. Both swelling powers of corn and red bean starch were increased rapidly from $70^{\circ}C$ to $90^{\circ}C$. 7, Gelatinization of corn and red bean were 75.6 and $61.8^{\circ}C$. 8. Brabender hot-paste viscosities of corn at 6% and 8% showed the similar amylogrm patterns with peak viscosity. And red bean had no peak viscosity. 9. The difference of sensory characteristics for ‘Mook’ and kidney bean & red bean starch gels was significant.
Roy, Swapan Kumar;Kwon, Soo Jeong;Cho, Seong-Woo;Kamal, Abu Hena Mostafa;Kim, Sang-Woo;Sarker, Kabita;Jeong, Hae-Ryong;Lee, Moon-Soon;Chung, Keun-Yook;Woo, Sun-Hee
Proceedings of the Korean Society of Crop Science Conference
/
2017.06a
/
pp.128-128
/
2017
Copper (Cu) is very toxic to plant cells due to its inhibitory effects on many physiological and biochemical processes. In spite of its potential physiological and economic significance, molecular characterization after Cu stress has so far been grossly overlooked in sorghum. To explore the molecular alterations that occur in response to copper stress, the present study was executed in ten-day-old Cu-exposed leaves of sorghum seedlings. The growth of shoots was markedly reduced, and ionic alterations were prominently observed in the leaves when the seedlings were exposed to different concentrations (0, 100, and $150{\mu}M$) of $CuSO_4$. Using two-dimensional gels with silver staining, 643 differentially expressed protein spots (${\geq}1.5-fold$) were identified as either significantly increased or reduced in abundance. Of these spots, a total of 24 protein spots (${\geq}1.5-fold$) from Cu-exposed sorghum leaves were successfully analyzed by MALDI-TOF-TOF mass spectrometry. Of the 24 differentially expressed proteins from Cu-exposed sorghum leaves, a total of 13 proteins were up-regulated, and 11 proteins were down-regulated. The abundance of most identified protein species, which function in carbohydrate metabolism, stress defense, and protein translation, was significantly enhanced, while that of another protein species involved in energy metabolism, photosynthesis and growth and development were severely reduced. The resulting differences in protein expression patterns together with related morpho-physiological processes suggested that these results could help to elucidate plant adaptation to Cu stress and provide insights into the molecular mechanisms of Cu responses in $C_4$ plants. The over-expression of GAPDH plays a significant role in assisting Sorghum bicolor to attenuate the adverse effects of oxidative stress caused by Cu, and the proteins involved in resistance to stress helped the sorghum plants to tolerate high levels of Cu.
Makgeolli mashes that were brewed using five different commercial nuruks (fermentation starters) were investigated for changes in physicochemistry, microbial diversity, and biogenic amine (BA) production. Mash A brewed with the nuruk (Gaeryang-nuruk) had the highest level of alcohol concentration and the greatest number of yeast cells, whereas mash E had the greatest number of bacterial cells. Only three biogenic amines were detected in the makgeolli mashes: tyramine, putrescine, and cadaverine. Using a PCR-DGGE technique, we observed that mash E had the highest BA production, and had the greatest number of bands on the denaturing gradient gels. We also observed that the numbers of bacterial cells correlated significantly with the putrescine and the total BA content, and that the BA content correlated significantly with the color values (L, a, b). This study shows that the quality of a makgeolli can depend on the type of nuruk. Therefore, we suggest that the quality management of makgeolli should start with the stage of nuruk manufacture.
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