• Journal of Microbiology and Biotechnology
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• v.17 no.2
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• pp.207-217
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• 2007

The Saccharomyces0 cerevisiae KNU5377 strain, which was isolated from spoilage in nature, has the ability to convert biomass to alcohol at high temperatures and it can resist against various stresses [18, 19]. In order to understand the defense mechanisms of the KNU5377 strain under menadione (MD) as oxidative stress, we used several techniques for study: peptide mass fingerprinting (PMF) by matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometry (MS) followed by two-dimensional (2D) gel electrophoresis, liquid chromatography electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS), and surface-enhanced laser desorption ionization-time of flight (SELDI-TOF) technology. Among the 35 proteins identified by MALDI-TOF MS, 19 proteins including Sod1p, Sod2p, Tsa1p, and Ahp1p were induced under stress condition, while 16 proteins were augmented under normal condition. In particular, five proteins, Sod1p, Sod2p, Ahp1p, Rib3p, Yaf9p, and Mnt1p, were induced in only stressed cells. By LC-ESI-MS/MS analysis, 37 proteins were identified in normal cells and 49 proteins were confirmed in the stressed cells. Among the identified proteins, 32 proteins were found in both cells. Five proteins including Yel047cp and Met6p were only upregulated in the normal cells, whereas 17 proteins including Abp1P and Sam1p were elevated in the stressed cells. It was interesting that highly hypothetical proteins such as Ynl281wp, Ygr279cp, Ypl273wp, Ykl133cp, and Ykr074wp were only expressed in the stressed cells. SELDI-TOF analysis using the SAX2 and WCX2 chips showed that highly multiple-specific protein patterns were reproducibly detected in ranges from 2.9 to 27.0 kDa both under normal and stress conditions. Therefore, induction of antioxidant proteins, hypothetical proteins, and low molecular weight proteins were revealed by different proteomic techniques. These results suggest that comparative analyses using proteomics might contribute to elucidate the defense mechanisms of KNU5377 under MD stress.

• Reproductive and Developmental Biology
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• v.33 no.3
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• pp.183-187
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• 2009

This study analyzed change of proteins in granulosa cells during the porcine follicuar development by proteomics techniques. Granulosa cells of the follicles, of which the diameter is $2{\sim}4\;mm$ and $6{\sim}10\;mm$, were collected from ovary of slaughtered pig that each follicle of diameter $1{\sim}4\;mm$ and $6{\sim}10\;mm$. We extracted glanulosa cell proteins by M-PER Mammalian Protein Extraction Reagent. Proteins were refined by clean-up kit and quantified by Bradford method until total protein was $200{\mu}l$. Immobilized pH gradient(IPG) strip used 18 cm, $3{\sim}10\;NL$. SDS-PAGE used 10% acrylamide gel. After silver staining, Melanie 7 and naked eye test were used for spot analyzation. Increasing proteins in glanulosa cell of $6{\sim}10\;mm$ follicle were 7 spots. This spots were analyzed by MALDI-TOF MS and searched on NCBInr. In results, 7 spots were similar to zinc/ling finger protein 3 precursor (RING finger protein 203), angiomotin, heat shock 60 kDa protein 1 (chaperonin) isoform 1 (HSP60), similar to transducin-like enhancer protein 1 (TLE 1), SH3 and PX domains 2A (SH3PXD2A). Those proteins were related with transfer between cells. Increase of proteins has an effect on follicular development.

• The Korean Journal of Zoology
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• v.35 no.3
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• pp.308-321
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• 1992

심근세포는 골격근 세포와 더불어 세포의 분화기작을 이해하기 위한 좋은 모델이라 할 수 있다. 본 연구에서는 심근세포의 배양중에 일어나는 심근 근원섬유형성과정에 대한 이해를 위하여 SDS- 및 two-dimensional gel electrophoresis와 immunofluorescent 및 immunoelectron microscopy를 수단으로 이를 검토하였다. 배양과정에서 나타나는 주요한 단백질 중에서 SDS-PAGE에 의해 209 kDa와 53 kDa 및 46 kDa band가 각각 막분획 및 세포질 분획 단백질에서 배양과정동안에 가장 현저한 양상을 보였다. 특히 배양 3일과 4일의 양상에서 뚜렷한 변화를 나타내어 이들의 OD 격은 배양 96시간에서의 막분획에서 0.38(209 kDa)와 0.52(53 kDa) 및 배양 72시간에서의 세포질 분획에서 0.34(46 kDa)로 각각 최고치를 나타내었다. 또한 이차원 전기영동상의 양상에서도 세포질 및 막분획 단백질의 전반적인 전개양상이 배양 96시간에서 가장 두드러짐으로써 결국 배양 72시간 및 96시간대에서 심근 모세포의 분화와 관련된 일련의 단백질 합성과정이 가장 활발하다는 것을 알 수 있었다. 배양중에 발현되는 actin에 대한 형광현미경적 분석에 의해, actin은 세포내에서 불균등하게 분포하였고 근원섬유가 형성되는 시기인 배양 96시간에 세포외곽부에서 나타나는 다소 강한 염색성을 관찰할 수 있었다. 심근세포의 근원섬유는 전자현미경 관찰에 의해 primitive forms로 배양 96시간에 출현함을 알 수 있었으며 이 시기의 근원섬유의 형태는 여러 구성대(A, H and M bands)가 미약하나 1대는 비교적 뚜렷하였다. 따라서 심근세포의 T근원섬유형성과정에서 관찰되는 뚜렷한 단백질양상의 변화와 근원섬유의 출현시기 및 구성과정간에는 중요한 관련성이 있으며 이러한 과정에서 actin은 세포질내에서 불균등한 분포를 나타낸다는 것을 알 수 있었다.

• Animal cells and systems
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• v.15 no.2
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• pp.147-154
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• 2011

Animal cell cultures generally require a nutrient-rich medium supplemented with animal serum. Adult bovine serum contains a variety of nutrients including inorganic minerals, vitamins, salts, proteins and lipids as well as growth factors that promote animal cell growth. To evaluate the potential use of gender-specific bovine serum (GSBS) for cell culture, the biochemical properties of male serum (MS), female serum (FS) and castrated-male serum (CMS) were investigated. Overall, the chemical profile of GSBS was similar to that of bovine references except for glucose, creatine kinase, lactate dehydrogenase and potassium. FS showed elevated total protein and sodium concentrations compared to MS and CMS. Proteins present in MS, FS and CMS but absent in fetal bovine serum (FBS) were selected by two-dimensional gel electrophoresis and identified by peptide mass fingerprinting. Some of the identified proteins are known to be involved in immune responses and the others have unknown physiological roles. Moreover, it was found that some proteins such as alpha-2-macroglobulin appeared to be gender-specific with higher contents in FS. Insulin and testosterone was significantly higher in MS, and $17{\beta}$-estradiol and estrone were higher in FS, as compared to the other sera. Taken together, the results indicate that each GSBS has a different ratio of components. Differences in serum constituents may affect cell cultures in a different manner and could be beneficial, depending on the specific aim of cell cultures.

• Applied Chemistry for Engineering
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• v.30 no.1
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• pp.23-28
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• 2019

We report a supercapacitor cable, which consists of three-dimensional (3D) porous graphene coated onto the surface of carbon thread. The 3D porous framework of graphene was constructed by microwave-activated process using a graphene oxide-coated carbon thread. The use of microwave irradiation enabled to convert graphene oxide into reduced graphene oxide without any reducing agents and activate graphene sheets into exfoliated and porous graphene sheets. Combining two wire electrodes with a polymer gel electrolyte successfully completed supercapacitor device in a form of cable construction. The supercapacitor cables were highly flexible, and thus can be transformed into various shapes of devices and be integrated into textile items. A high area-capacitance of 38.1 mF/cm was obtained at a scan rate of 10 mV/s. This capacitance was retained 88% of its original value at 500 mV/s. The cycle life was also demonstrated by repeating a charge/discharge process during 10,000 cycles even under bent states, showing a high capacitance retention of 96.5%.

• Applied Microscopy
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• v.49
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• pp.3.1-3.7
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• 2019

Graphene, which is one of the most promising materials for its state-of-the-art applications, has received extensive attention because of its superior mechanical properties. However, there is little experimental evidence related to the mechanical properties of graphene at the atomic level because of the challenges associated with transferring atomically-thin two-dimensional (2D) materials onto microelectromechanical systems (MEMS) devices. In this study, we show successful dry transfer with a gel material of a stable, clean, and free-standing exfoliated graphene film onto a push-to-pull (PTP) device, which is a MEMS device used for uniaxial tensile testing in in situ transmission electron microscopy (TEM). Through the results of optical microscopy, Raman spectroscopy, and TEM, we demonstrate high quality exfoliated graphene on the PTP device. Finally, the stress-strain results corresponding to propagating cracks in folded graphene were simultaneously obtained during the tensile tests in TEM. The zigzag and armchair edges of graphene confirmed that the fracture occurred in association with the hexagonal lattice structure of graphene while the tensile testing. In the wake of the results, we envision the dedicated preparation and in situ TEM tensile experiments advance the understanding of the relationship between the mechanical properties and structural characteristics of 2D materials.

• Mycobiology
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• v.49 no.6
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• pp.604-606
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• 2021

In our ongoing search for new secondary metabolites from fungal strains, one novel compound (1) and nine known compounds (2-10) were isolated from the EtOAc-soluble layer of the culture broth of Panus rudis. The culture broth of P. rudis was extracted in acetone and fractionated by solvent partition; column chromatography using silica gel, Sephadex LH-20, and Sephadex G-10; MPLC; and HPLC. The structures of isolated compounds were elucidated by one- and two-dimensional NMR and LC-ESI-mass measurements. One new compound, panepoxydiol (1), and nine known compounds, (E)-3-(3-hydroxy-3-methylbut-1-en-1-yl)-7-oxabicyclo[4.1.0]hept-3-ene-2,5-diol (2), isopanepoxydone (3), neopanepoxydone (4), panepoxydone (5), panepophenanthrin (6), 4-hydroxy-2,2-dimethyl-6-methoxychromane (7), 6-hydroxy-2,2-dimethyl-3-chromen (8), 2,2-dimethyl-6-methoxychroman-4-one (9), 3,4-dihydroxy-2,2-dimethyl-6-methoxychromane (10), were isolated from the culture broth of P. rudis. This is the first report of isolation of a new compound panepoxydiol (1) and nine other chemical constituents (2-5, 7-10) from the culture broth of P. rudis.

• Journal of Forest and Environmental Science
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• v.26 no.2
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• pp.83-94
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• 2010

Chinese fir (Latin name: Cunninghaimia lanceolata) is one of the major commercial coniferous trees. Most of Chinese fir forests are managed in successive rotation sites, which lead productivity to decline. Autotoxicity is the important reason for soil degradation of Chinese fir plantation, especially, phenolic acids are considered as the major allelopathic toxins which induce autotoxicity in Chinese fir rotation stands. We performed here proteomic approach to investigate the response of proteins in Chinese fir leaves to salicylic acid. The tube plantlets of Chinese fir clone were treated with 120 mg/L salicylic acid for 1, 3 and 5th day. 2-DE, coupled with MALDI-TOF-TOF/MS, was used to separate and identify the responsive proteins. We found 12, 7, and 12 candidate protein spots that were up- or down-regulated by at least 2.5 fold after 1, 3, and 5th day of the stress, respectively. Of these protein spots, 16 spots were identified successfully. According to the putative physiological functions, these proteins were categorized into five classes (1) the proteins involved in protein stability and folding, including 26S proteome, Grp78, Hsp70, Hsp90 and PPIase; (2) the protein involved in photosynthesis and respiration, including OEC 33 kDa subunit, GAPDH; (3) the protein related to cell endurance to acid, F-ATPase; (4) the protein related to cytoskeleton, tubulin; (5) the protein related to protein translation: prolyl-tRNA synthetase. These results give new insights into autotoxic substance stress response in Chinese fir leaves and provide preliminary footprints for further studies on the molecular signal mechanisms induced by the stress.

• Mycobiology
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• v.50 no.5
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• pp.366-373
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• 2022

Regulation of proper gene expression is important for cellular and organismal survival, maintenance, and growth. Abnormal gene expression, even for a single critical gene, can thwart cellular integrity and normal physiology to cause diseases, aging, and death. Therefore, gene expression profiling serves as a powerful tool to understand the pathology of diseases and to cure them. In this study, the difference in gene expression in Flammulina velutipes was compared between the wild type (WT) mushroom and the mutant one with clogging phenomenon. Differentially expressed transcripts were screened to identify the candidate genes responsible for the mutant phenotype using the DNA microarray analysis. A total of 88 genes including 60 upregulated and 28 downregulated genes were validated using the real-time quantitative PCR analysis. In addition, proteomic differences between the WT and mutant mushroom were analyzed using two-dimensional gel electrophoresis and matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF). Interestingly, the genes identified by these genomic and proteomic analyses were involved in stress response, translation, and energy/sugar metabolism, including HSP70, elongation factor 2, and pyruvate kinase. Together, our data suggest that the aberrant expression of these genes attributes to the mutant clogging phenotype. We propose that these genes can be targeted to foster normal growth in F. velutipes.

• Food Science of Animal Resources
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• v.44 no.3
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• pp.533-550
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• 2024

Peptides with bioactive effects are being researched for various purposes. However, there is a lack of overall research on pork-derived peptides. In this study, we reviewed the process of obtaining bioactive peptides, available analytical methods, and the study of bioactive peptides derived from pork. Pepsin and trypsin, two representative protein digestive enzymes in the body, are hydrolyzed by other cofactors to produce peptides. Bicinchoninic acid assay, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, chromatography, and in vitro digestion simulation systems are utilized to analyze bioactive peptides for protein digestibility and molecular weight distribution. Pork-derived peptides mainly exhibit antioxidant and antihypertensive activities. The antioxidant activity of bioactive peptides increases the accessibility of amino acid residues by disrupting the three-dimensional structure of proteins, affecting free radical scavenging, reactive oxygen species inactivation, and metal ion chelating. In addition, the antihypertensive activity decreases angiotensin II production by inhibiting angiotensin converting enzyme and suppresses blood pressure by blocking the AT1 receptor. Pork-derived bioactive peptides, primarily obtained using papain and pepsin, exhibit significant antioxidant and antihypertensive activities, with most having low molecular weights below 1 kDa. This study may aid in the future development of bioactive peptides and serve as a valuable reference for pork-derived peptides.