• Molecules and Cells
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• v.22 no.1
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• pp.119-125
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• 2006

The rat is an accepted model for studying human psychiatric/neurological disorders. We provide a protocol for total soluble protein extraction using trichloroacetic acid/acetone (TCA/A) from rat (female) whole brain, 10 brain regions and the pituitary gland, and show that two-dimensional gel electrophoresis (2-DGE) using precast immobilized pH (4-7) gradient (IPG) strip gels (13 cm) in the first dimension yields clean silver nitrate stained protein profiles. Though TCA/A precipitation may not be "ideal", the important choice here is the selection of an appropriate lysis buffer (LB) for solubilizing precipitated proteins. Our results reveal enrichment of protein spots by use of individual brain regions rather than whole brain, as well as the presence of differentially expressed spots in their proteomes. Thus individual brain regions provide improved protein coverage and are better suited for differential protein detection. Moreover, using a phosphoprotein-specific dye, ingel detection of phosphoproteins was demonstrated. Representative high-resolution silver nitrate stained proteome profiles of rat whole brain total soluble protein are presented. Shortcomings apart (failure to separate membrane proteins), gel-based proteomics remains a viable option, and 2-DGE is the method of choice for generating high-resolution proteome maps of rat brain and brain regions.

• Proceedings of the Microbiological Society of Korea Conference
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• 2008.05a
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• pp.118-118
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• 2008

The phylogenetic diversity of microbial communities in calcareous mats from Spearfish Creek, a freshwater stream located in the Black Hills of South Dakota, was examined using PCR-based 16S rDNA sequence analysis. In this study, two types of calcareous mats were compared: mature mats formed on the natural substrate of rock surfaces and developing mats on an artificial substrate of glass slides. Among 63 unique isolates from a clone library of 16S rRNA genes from mature mat samples, there were 8 phyla of Bacteria represented. The predominant phylum was Proteobacteria (48%), with the $\beta$ subclass being the largest group. Denaturing gradient gel electrophoresis (DGGE) analysis of 16S rRNA genes from slide samples collected at intervals for four months showed considerable diversity of the microbial community from the earliest stages of community development. Amplicons isolated from DGGE gels and sequenced indicated that community succession has occurred without increasing microbial diversity. However, light microscopic analysis revealed a significant increase in microbial cell density throughout the community development. Scanning electron microscopy of mat samples provides evidence that diatoms are also important members of calcareous mat communities.

• Reproductive and Developmental Biology
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• v.33 no.4
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• pp.243-248
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• 2009

To examine the differential expression of proteins during the cycling (70~80% confluences) and G0/G1 (full confluences) phases in porcine fetal fibroblast cells, we used a global proteomics approach by 2-D gel electrophoresis (2-DE) and MALDI-TOF-MS. Cycling cell were harvested at approximately 70% to 80% confluent state while cells in G0/G1 phase were recovered after maintenance of a confluent state for 48 hr. Cellular proteins with isoelectric points ranging between 3.0~10.0, were analyzed by 2-DE with 2 replicates of each sample. A total of approximately 700 spots were detected by 2.D gels stained with Coomassie brilliant blue. On comparing the cell samples obtained from the cycling and G0/G1 phases, a total of 13 spots were identified as differentially expressed proteins, of which 8 spots were up-regulated in the cycling cell and 5 were up-regulated in the G0/G1 phase. Differentially expressed proteins included K3 keratin, similar to serine protease 23 precursor, protein disulfide-isomerase A3, microsomal protease ER-60, alpha-actinin-2, and heat-shock protein 90 beta. The identified proteins were grouped on the basis of their basic functions such as molecular binding, catabolic, cell growth, and transcription regulatory proteins. Our results show expression profiles of key proteins in porcine fetal fibroblast cells during different cell cycle status.

• Journal of Pharmaceutical Investigation
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• v.36 no.4
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• pp.253-262
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• 2006

This study was performed to investigate the effects of treatment for dermatitis using the herbal gel preparations. Scutellariae Radix(SR) and Zingiberis Rhizoma(ZR) were used for the purpose of herbal preparations. Baicalin, baicalein are the ingredients of SR, having biological effects like anti-inflammatory, anti-oxidative, anti-bacterial and antiallergic action. 6-Gingerol is one of the ingredients of ZR having biologicai effects like anti-inflammatory and analgesic action. The three types of hydrogels(SRE, SRH, SZH) were formulated with Carbopol 940, Labrasol, Triethanolamine etc. Baicalin was hydrolysed to baicalein by $\beta$-glucuronidase for the purpose to increase rate of skin permeation. Content of ingredients by HPLC determination, rate of skin permeation using Franz type diffusion cell, anti-oxidative activity for the free radical, hydroxyl radical, superoxide, anti-inflammatory by using carrageenan injection, efficacy on the dermatitis induced by 2,4-dinitro-chlorobenzene(DNCB) were experimented. Baicalein showed higher permeability than baicalin, so it is considered that baicalein was more suitable form than baicalin for transdermal absorption by its lipophilic property. In the anti-oxidative experiments, SZH gel was the most effective scavenging activity than the other gels in all experiments. In anti-inflammatory test, SRM and SZH gel more decreased edma rapidly than SRE gel. In case of using SZH gel, treatment period for the dermatitis was more declined than that of other gel groups. These results suggests that the SZH hydrogel could be suitable preparations for the treatment of dermatitis.

• Reproductive and Developmental Biology
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• v.36 no.2
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• pp.121-131
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• 2012

After spermatogenesis, spermatozoa come in contact with fluids in the epididymis where they mature. During ejaculation, spermatozoa are mixed with secretions from prostate gland, vesicular glands, and bulbourethral glands. During natural mating, seminal plasma is deposited in the female reproductive tract eliciting various physiological and immunological responses. With the advances in proteomics, the components of seminal plasma have been identified and the information may be valuable in identifying markers for fertility. Components of seminal plasma that affect fertility have been discovered and the mechanism of action of these factors has been determined. The objective of this study was to determine the specific seminal plasma proteins from Korean native cattle, Hanwoo, and Korean native brindle cattle (KNBC) with the long term goal of improving fertilization rate. After SDS-PAGE and 2-dimensional gel electrophoresis, proteins were identified by Q-ToF analysis. They include plasma serine protease inhibitor precursor and platelet-activating factor acetylhydrolase after SDS-PAGE. Number and density of the spots in 2-dimensional gels were higher in KNBC than Hanwoo. Proteins identified from the paired spots of both breeds include chain A, bull seminal plasma PDC-109 Fibronectin Type II module, BSP-30 kDa precursor, and Spermadhesin Z13 or its precursor. Interestingly, some proteins were identified from multiple spots. The functional differences of these diverse forms of the proteins may require further studies. With their previously reported roles in sperm capacitation by these proteins, the studies on the mechanism of action, ligand interaction and the variation in the genome may help improving fertility in cattle.

• Korean journal of food and cookery science
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• v.18 no.2
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• pp.157-163
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• 2002

The gelatinization and retrogradation properties of rice starch added with dietary fibers from sikhe were investigated. The pasting temperature and setback of starch measured by using a rapid visco analyzer were increased by the addition of the fiber, whereas the peak viscosity, trough viscosity, end viscosity, breakdown and consistency were lowered. The onset temperature of starch measured by a DSC was increased, but the enthalpy of gelatinization of the starch was decreased as the addition of fiber increased. The melting peak temperature of the retrograded starch gels was 41∼46$\^{C}$ as measured by a DSC. The melting enthalpy of the control retrograded starch gel was increased while storing for 1, 3 and 7 days at 4$\^{C}$, but that of containing sikhe fiber showed no differences.

• Journal of Embryo Transfer
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• v.25 no.4
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• pp.281-286
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• 2010

This study was to evaluate the protein profile of seminal plasma using 2-DE in Hanwoo. Seminal plasma was harvested from five mature Hanwoo, and seminal plasma protein was extracted by M-PER Mammalian Protein Extraction Reagent. Proteins were refined by clean-up kit and quantified by Bradford method until total protein was $300\;{\mu}l$. Immobilized pH gradient (IPG) strip was used 18 cm and 3~11 NL. SDS-PAGE was used 12% acrylamide gel. Each gels were visualized by comassie brilliant blue and silver staining. These spots were analyzed by MALDI-TOF MS and searched on NCBInr. The result, 20 proteins of 36 protein spots were searched through peptide sequencing on the NCBInr. 8 proteins profiled by 2-DE were proved through previous bovine studies and the name of each protein was albumin, nucleobindin, clusterin, TIMP-2, spermadhesin Z13, spermadhesin-1 and BSP proteins (BSP 30 kDa and BSP A1/A2). 12 new proteins were ATP synthase, protein MAK16 homolog, Transmembrane protein 214, E3 ubiquitin-protein ligase BRE1A, dual serine/threonine and tyrosine protein kinase, tissue factor pathway inhibitor 2, alpha-actinin-4, RUN domain-containing protein 3B, catenin alpha-1, protein-glutamine gamma-glutamyltransferase 2, plakophilin-1 and inter-alpha-trypsin inhibitor heavy chain H1 has not been previously described in the bovine seminal plasma study. These proteins may be contribute to define the type of proteins affecting fertility of male and improve the fertilizing ability of semen in Hanwoo.

• Microbiology and Biotechnology Letters
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• v.20 no.4
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• pp.451-458
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• 1992

Pervaporation which is capable of removing ethanol selectively was adopted to reduce the ethanol inhibition and in situ recovery of ethanol in ethanol fermentation, The composite membrane made of silicone and polysulfone was used to separate the ethanol selectively. The ethanol selectivity of the membrane was about 4 and the total flux was 300 g/m2 h at 301:: and 10 mmHg for 25 g/l of feed concentration. Saccharomyces cerevisiae entrapped within Ca-alginate gels was employed for ethanol fermentations in a fluidized-bed bioreactor. The pervaporation membrane unit and fluidized-bed bioreactor were combined into one system. The proposed model equations for the combined system showed good accordances with the experimental results. It was found from the simulation results that the ethanol concentration in the broth for the combined system was lower than that for the continuous fermentation system without a membrane unit. Ethanol productivity can be thus increased by employing the combined system.

• Korean Journal of Food Science and Technology
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• v.27 no.5
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• pp.690-696
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• 1995

Properties of dry milled rice flours made of paddy with different moisture content (12.6, 18.4 and 24.4%) were compared with those of wet milled rice flour. Among dry milled rice flour, the higher moisture content of paddy the finer particle size and the more distinct shape of starch were observed. As the moisture content of paddy increased, L (lightness) value and water absorption index of rice flour were increased, while setback in amylogram and water soluble index were decreased. Lower gel consistency and gel strength were found in dry milled rice flours than in wet milled rice flour, while no significant difference was found among dry milled rice flours. Enthalpy for melting crystalline of retrograded gels was higher in dry milled rice flours made of paddy with 24.4% moisture content than in other dry milled rice flours. Wet milled rice flour had lower retrogradation enthalphy than dry milled rice flours.

• Korean Journal of Food Science and Technology
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• v.22 no.7
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• pp.812-816
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• 1990

This study describes the sorbitol production with permeabilized cells of Zymomonas mobilis immobilized in Ca-alginate. Toluene treated cells lose activity of glucose-fructose oxidoreductase due to the leaking of enzyme from the cells. To prevent this leakage, the permeabilized cells were treated with 0.25% glutaraldehyde by stirring for 1 h at room temperature. A continuous process with glutaraldehyde treated cells was developed and no significant reduction in the degree of conversion occurred during 210 h operation. The productivities were estimated to be about $7.2{\sim}7.5\;g/l-h$ for sorbitol at dilution rate $0.18\;h^{-1}$.