• Title/Summary/Keyword: Functional characterization

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Immobilization of As and Pb in Contaminated Soil Using Bead Type Amendment Prepared by Iron NanoparticlesImpregnated Biochar (철 나노 입자가 담지된 바이오차 기반 비드 형태 안정화제를 이용한 비소 및 납 오염토양의 안정화)

  • Choi, Yu-Lim;Kim, Dong-Su;Kang, Tae-Jun;Yang, Jae-Kyu;Chang, Yoon-Young
    • Journal of Environmental Impact Assessment
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    • v.30 no.4
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    • pp.247-257
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    • 2021
  • In this work, Iron Nano-Particles Impregnated BioChar/bead (INPBC/bead) soil amendment was developed to increase biochar's reactivity to As in soil and preventing possible wind loss. Prior to preparation of INPBC/bead, INPBC was produced utilizing lignocellulosic biomass and Fe(III) solution in a hydrothermal method, followed by a calcination process. Then, the bead type amendment, INPBC/bead was produced by cross-linking reaction of alginate with INPBC. FT-IR, XRD, BET, and SEM-EDS analyses were utilized to characterize the as-synthesised materials. The particle size range of INPBC/bead was 1-4 mm, and different oxygen-containing functional groups and Fe3O4 crystalline phase were produced on the surface of INPBC/bead, according to the characterization results. The soil cultivation test was carried out in order to assess the stabilization performance of INPBC/bead utilizing As and Pb-contaminated soil obtained from an abandoned mining location in South Korea. After 4 weeks of culture, TCLP and SPLP extraction tests were performed to assess the stabilization efficacy of the amendment. The TCLP and SPLP findings revealed that raising the application ratio improved stabilizing efficiency. The As stabilization efficiency was determined to be 81.56 % based on SPLP test findings for a 5% in (w/w) INPBC/bead treatment, and the content of Pb in extracts was reduced to the limit of detection. According to the findings of this study, INPBC/bead that can maintain pH of origin soil and minimize wind loss might be a potential amendment for soil polluted with As and heavy metals.

Analysis of 16S rRNA gene sequencing data for the taxonomic characterization of the vaginal and the fecal microbial communities in Hanwoo

  • Choi, Soyoung;Cha, Jihye;Song, Minji;Son, JuHwan;Park, Mi-Rim;Lim, Yeong-jo;Kim, Tae-Hun;Lee, Kyung-Tai;Park, Woncheoul
    • Animal Bioscience
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    • v.35 no.11
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    • pp.1808-1816
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    • 2022
  • Objective: The study of Hanwoo (Korean native cattle) has mainly been focused on meat quality and productivity. Recently the field of microbiome research has increased dramatically. However, the information on the microbiome in Hanwoo is still insufficient, especially relationship between vagina and feces. Therefore, the purpose of this study is to examine the microbial community characteristics by analyzing the 16S rRNA sequencing data of Hanwoo vagina and feces, as well as to confirm the difference and correlation between vaginal and fecal microorganisms. As a result, the goal is to investigate if fecal microbiome can be used to predict vaginal microbiome. Methods: A total of 31 clinically healthy Hanwoo that delivered healthy calves more than once in Cheongju, South Korea were enrolled in this study. During the breeding season, we collected vaginal and fecal samples and sequenced the microbial 16S rRNA genes V3-V4 hypervariable regions from microbial DNA of samples. Results: The results revealed that the phylum-level microorganisms with the largest relative distribution were Firmicutes, Actinobacteria, Bacteroidetes, and Proteobacteria in the vagina, and Firmicutes, Bacteroidetes, and Spirochaetes in the feces, respectively. In the analysis of alpha, beta diversity, and effect size measurements (LefSe), the results showed significant differences between the vaginal and fecal samples. We also identified the function of these differentially abundant microorganisms by functional annotation analyses. But there is no significant correlation between vaginal and fecal microbiome. Conclusion: There is a significant difference between vaginal and fecal microbiome, but no significant correlation. Therefore, it is difficult to interrelate vaginal microbiome as fecal microbiome in Hanwoo. In a further study, it will be necessary to identify the genetic relationship of the entire microorganism between vagina and feces through the whole metagenome sequencing analysis and meta-transcriptome analysis to figure out their relationship.

Isolation of Simiduia sp. SH-2 and Characterization of Its β-Agarase (한천분해세균 Simiduia sp. SH-2 균주의 분리 및 β-agarase의 특성조사)

  • Lee, Dong-Geun;Kim, Geun-Dae;Lee, Sang-Hyeon
    • Journal of Life Science
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    • v.32 no.10
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    • pp.778-783
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    • 2022
  • This study isolated a new agarase-producing bacterium and characterized its agarase. A new agar-degrading strain was isolated from the seashore of Namhae in Gyeongnam province, Korea, and was purely cultured using the Marine Agar 2216 media. The isolated bacterium was identified as Simiduia sp. SH-2 after 16S rRNA gene sequencing. The crude agarase was obtained from the culture medium of the Simiduia sp. SH-2 strain, and the agar-degrading activity was measured. The highest level of activity of the Simiduia sp. SH-2-derived agar-degrading enzyme was 625 U/l. Agar degradation activity was most significant at 40℃ and pH 7.0. Compared to the activity at 40℃, the relative activity was 31% at 20℃ and 71% at 30℃. Compared to the activity at pH 7.0, the relative activity was 94% and 89% at pH 6.0 and pH 8.0, respectively. Residual activity was greater than 96% after exposure to 20℃ and 30℃ for 2 hr and more than 49% after exposure to 40℃ for 2 hr. Simiduia sp. SH-2 was identified as a strain producing β-agarase that creates neoagarooligosaccharides, such as neoagarotetraose and neoagarohexaose. Therefore, the Simiduia sp. SH-2 strain and its β-agarase are expected to be useful functional material producers in the food, cosmetic, and pharmaceutical industries.

Functional characterization of naturally-occurring constitutively activating/inactivating mutations in equine follicle-stimulating hormone receptor

  • Byambaragchaa, Munkhzaya;Ahn, Tae-Young;Choi, Seung-Hee;Kang, Myung-Hwa;Min, Kwan-Sik
    • Animal Bioscience
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    • v.35 no.3
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    • pp.399-409
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    • 2022
  • Objective: Follicle-stimulating hormone (FSH) is the central hormone involved in mammalian reproduction, maturation at puberty, and gamete production that mediates its function by control of follicle growth and function. The present study investigated the mutations involved in the regulation of FSH receptor (FSHR) activation. Methods: We analyzed seven naturally-occurring mutations that were previously reported in human FSHR (hFSHR), in the context of equine FSHR (eFSHR); these include one constitutively activation variant, one allelic variant, and five inactivating variants. These mutations were introduced into wild-type eFSHR (eFSHR-wt) sequence to generate mutants that were designated as eFSHR-D566G, -A306T, -A189V, -N191I, -R572C, -A574V, and -R633H. Mutants were transfected into PathHunter EA-parental CHO-K1 cells expressing β-arrestin. The biological function of mutants was analyzed by quantitating cAMP accumulation in cells incubated with increasing concentrations of FSH. Results: Cells expressing eFSHR-D566G exhibited an 8.6-fold increase in basal cAMP response, as compared to that in eFSHR-wt. The allelic variation mutant eFSHR-A306T was not found to affect the basal cAMP response or half maximal effective concentration (EC50) levels. On the other hand, eFSHR-D566G and eFSHR-A306T displayed a 1.5- and 1.4-fold increase in the maximal response, respectively. Signal transduction was found to be completely impaired in case of the inactivating mutants eFSHR-A189V, -R572C, and -A574V. When compared with eFSHR-wt, eFSHR-N191I displayed a 5.4-fold decrease in the EC50 levels (3,910 ng/mL) and a 2.3-fold decrease in the maximal response. In contrast, cells expressing eFSHR-R633H displayed in a similar manner to that of the cells expressing the eFSHR-wt on signal transduction and maximal response. Conclusion: The activating mutant eFSHR-D566G greatly enhanced the signal transduction in response to FSH, in the absence of agonist treatment. We suggest that the state of activation of the eFSHR can modulate its basal cAMP accumulation.

Identification and Characterization of Secreted Phosphoprotein 2 as a Novel Bioactive Protein for Myocardial Differentiation (심근세포로의 분화에 관여하는 새로운 생리활성 단백질 SPP2의 발굴)

  • Sejin Jeon
    • Journal of Life Science
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    • v.33 no.1
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    • pp.64-72
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    • 2023
  • Despite several advances in identification of cardiac transcription factors, there are still needs to find new bioactive molecules that promote cardiomyogenesis from stem cells to highly efficient myocardial differentiation. We analyzed Illumina expression microarray data of mouse embryonic stem cells (mESCs)-derived cardiomyocytes. 276 genes were upregulated (≥ 4fold) in mESCs-derived cardiomyocytes compared undifferentiated ESCs. Secreted phosphoprotein 2 (Spp2) is one of candidates and is known to inhibit bone morphogenetic protein 2 (BMP2) signal transduction as a pseudoreceptor for BMP2. However, its function in cardiomyogenesis is unknown. We confirmed that Spp2 expression increased during the differentiation into functional cardiomyocytes using mESCs, TC-1/Kh2 and E14. Interestingly, Spp2 secretion transiently increased 3 days after formation of embryoid bodies (EBs), indicating that the extracellular secretion of Spp2 is involved in the differentiation of ESCs into cardiomyocytes. To characterize Spp2, we performed experiments using the C2C12 mouse myoblast cell line, which has the property of shifting the differentiation pathway from myoblastic to osteoblastic by treatment with BMP2. Similar to the differentiation of ESCs, transcription of Spp2 increased as C2C12 myoblasts differentiated into myotubes. In particular, Spp2 secretion increased dramatically in the early stage of differentiation. Furthermore, treatment with Spp2-Flag recombinant protein promoted the differentiation of C2C12 myoblasts into myotubes. Taken together, we suggest a novel bioactive protein Spp2 that differentiates ESCs into cardiomyocytes. This may be useful for understanding the molecular pathways of cardiomyogenesis and for experimental or clinical promotion of stem cell therapy for ischemic heart diseases.

Preparation of Dual-functionalized Polymeric Membrane Electrolyte and Ni, Co-based Nanowire/MOF Array on Carbon Cloth for High-performance Supercapacitor (이중 기능 고분자 전해질 막의 제조 및 탄소 섬유에 니켈, 코발트 기반의 나노와이어/MOF 배열을 통한 고성능 슈퍼커패시터 연구)

  • Hye Jeong Son;Bong Seok Kim;Ji Min Kwon;Yu Bin Kang;Chang Soo Lee
    • Membrane Journal
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    • v.33 no.4
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    • pp.211-221
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    • 2023
  • This study presents a comprehensive study on the synthesis and characterization of PVI-PGMA/LiTFSI polymeric membrane electrolytes and CxNy-C flexible electrodes for energy storage applications. The dual-functional PVI-PGMA copolymer exhibited excellent ionic conductivity, with the PVI-PGMA73/LiTFSI200 membrane electrolyte achieving the highest conductivity of 1.0 × 10-3 S cm-1. The electrochemical performance of the CxNy-C electrodes was systematically investigated, with C3N2-C demonstrating superior performance, achieving the highest specific capacitance of 958 F g-1 and lowest charge transfer resistance (Rct) due to its highly interconnected hybrid structure comprising nanowires and polyhedrons, along with binary Co/Ni oxides, which provided abundant redox-active sites and facilitated ion diffusion. The presence of a graphitic carbon shell further contributed to the enhanced electrochemical stability during charge-discharge cycles. These results highlight the potential of PVI-PGMA/LiTFSI polymeric membrane electrolytes and CxNy-C electrodes for advanced energy storage devices, such as supercapacitors and lithium-ion batteries, paving the way for further advancements in sustainable and high-performance energy storage technologies.

Characterization of Carbamazepine-Imprinted Acorn Starch/PVA-Based Biomaterials (카바마제핀 각인 도토리 전분/PVA 기반 바이오소재의 특성)

  • Kyeong-Jung Kim;Ji-Hoon Kang;Bo-Gyeong Kim;Min‑Jin Hwang;Soon-Do Yoon
    • Applied Chemistry for Engineering
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    • v.35 no.3
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    • pp.192-199
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    • 2024
  • In this study, carbamazepine (CBZ) imprinted starch/PVA-based biomaterials were prepared by the casting method and UV irradiation, and their physicochemical properties, CBZ adsorption ability, and release properties were investigated. The surface properties of the prepared biomaterials were characterized using FE-SEM, while the stability of CBZ under UV irradiation and the functional groups of the biomaterials were characterized using FT-IR analysis. The adsorption properties of CBZ on the biomaterials were evaluated by binding isotherm and Scatchard plot. Results indicate that CBZ imprinted biomaterials possess a specific binding site of CBZ. To evaluate the applicability of the transdermal drug delivery system, the release properties of CBZ from prepared biomaterials using various pH buffers and artificial skin at 36.5 ℃ were investigated. Results indicated that the CBZ release at high pH was faster than at low pH. In addition, CBZ was released continuously for 12 h in the artificial skin test. The drug release mechanism of CBZ followed a pseudo-Fickian diffusion mechanism in buffer solution, whereas the release from artificial skin exhibited a non-Fickian diffusion mechanism.

Enzymatic characterization of Paenibacillus amylolyticus xylanases GH10 and GH30 for xylan hydrolysis (Paenibacillus amylolyticus 유래 xylanase GH10 및 GH30의 xylan 가수분해 특성)

  • Nam, Gyeong-Hwa;Jang, Myoung-Uoon;Kim, Min-Jeong;Lee, Jung-Min;Lee, Min-Jae;Kim, Tae-Jip
    • Korean Journal of Microbiology
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    • v.52 no.4
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    • pp.463-470
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    • 2016
  • The enzymatic degradation of xylans is the most versatile way to obtain the high value-added functional compounds or the fermentable sugars for renewable energy. The endo-${\beta}$-xylanases are the major enzymes which hydrolyze the internal ${\beta}$-1,4-linkages of xylan backbones to produce the mixtures of xylooligosaccharides including xylobiose and xylotriose. Among them, glucuronoxylanase GH30 can exclusively hydrolyze the internal ${\beta}$-1,4-linkages of xylans decorated with methylglucuronic acid branches. In the present study, two xylanolytic enzyme (PaXN_10 and PaGuXN_30) genes were cloned from Paenibacillus amylolyticus KCTC 3005, and expressed in Escherichia coli, respectively. PaXN_10 (38.7 kDa) belongs to the endo-${\beta}$-xylanases GH10 family, while PaGuXN_30 (58.5 kDa) is a member of glucuronoxylanase GH30. They share the same optimal reaction conditions at $50^{\circ}C$ and pH 7.0. Enzymatic characterization proposed that P. amylolyticus can utilize the hardwood glucuronoarabinoxylans via the cooperative actions of xylanases GH10 and GH30. The extracellular PaGuXN_30 is secreted into the medium and hydrolyzes glucuronoarabinoxylans to release a series of aldouronic acid mixtures with a methylglucuronic acid branch. The resultant products being transported into the microbial cell are successively degraded into the smaller xylooligosaccharides by the intracellular PaXN_10, which will be utilized for the cellular metabolism.

Characterization of Cigarette Smoke Extract (CSE)-induced Cell Death in Lung Epithelial Cells (폐상피세포에서 흡연추출물-유도성 세포사에 관한 연구)

  • Choi, Eun Kyung;Kim, Yun Seup;Park, Jae Seuk;Jee, Young Koo;Lee, Kye Young
    • Tuberculosis and Respiratory Diseases
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    • v.58 no.1
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    • pp.43-53
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    • 2005
  • Emphysema is characterized by air space enlargement and alveolar destruction. The mechanism responsible for the development of emphysema was thought to be protease/antiprotease imbalance and oxidative stress. A very recent study shows that alveolar cell apoptosis causes lung destruction and emphysematous changes. Thus, this study was performed to support the evidence for the role of apoptosis in the development of emphysema by characterizing cigarette smoke extract (CSE)-induced apoptosis in A549 (type II pneumocyte) lung epithelial cells. CSE induced apoptosis at low concentration (10% or less) and both apoptosis and necrosis at high concentration (20%). Apoptosis was demonstrated by DNA fragmentation using FACScan for subG1 fraction. Discrimination between apoptosis and necrosis was done by morphologic analysis using fluorescent microscopy with Hoecst 33342/propium iodide double staing and electron microscopy. Cytochrome c release was confirmed by using immunofluorescence with monoclonal anti-cytochrome c antibody. However, CSE-induced cell death did not show the activation of caspase 3 and was not blocked by caspase inhibitors. This suggests that CSE-induced apoptosis might be caspase-independent apoptosis. CSE-induced cell death was near completely blocked by N-acetylcystein and bcl-2 overexpression protected CSE-induced cell death. This results suggests that CSE might induce apoptosis through intracellular oxidative stress. CSE also activated p53 and functional knock-out of p53 using stable overexpression of HPV-E6 protein inhibited CSE-induced cell death. The characterization of CSE-induced cell death in lung epithelial cells could support the role of lung cell apoptosis in the pathogenesis of emphysema.

Proteome Characterization of Sorghum (Sorghum bicolor L.) at Vegetative Stage under Waterlogging Stress (토양 과습 조건하에서 수수 잎의 단백질 양상)

  • Yun, Min Heon;Jeong, Hae-Ryong;Yoo, Jang-Hwan;Roy, Swapan Kumar;Kwon, Soo-Jeong;Kim, Joo-Ho;Chun, Hyen Chung;Jung, Ki Yuol;Cho, Seong-Woo;Woo, Sun-Hee
    • Korean Journal of Plant Resources
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    • v.31 no.2
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    • pp.124-135
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    • 2018
  • The study was performed to explore the molecular changes in the vegetative stage (3-and 5-leaf) of sorghum under waterlogging stress. A total of 74 differentially expressed protein spots were analyzed using LTQ-FT-ICR MS. Among them, 12 proteins were up-regulated and 3 proteins were down-regulated. Mass spectrometry (MS) results showed that about 50% of the proteins involved in various metabolic processes. The level of protein expression of malate dehydrogenase and glyceraldehyde-3-phosphate dehydrogenase related to carbohydrate metabolic process increased in both 3 and 5-leaf stage under waterlogging stress. These proteins are known to function as antistress agents against waterlogging stress. The expression of oxygen-evolving enhancer protein 1 protein related to photosynthesis was slightly increased in the treated group than in the control group, however the expression level was increased in the 5-leaf stage compared to the 3-leaf stage. Probable phospholipid hydroperoxide glutathione peroxidase protein and superoxide dismutase protein related to response to oxidative stress showed the highest expression level in 5-leaf stage treatment. This suggests that the production of reactive oxygen species by the waterlogging stress was the most abundant in the 5-leaf treatment group, and the expression of the antioxidant defense protein was increased.