• Journal of Microbiology and Biotechnology
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• 제30권2호
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• pp.244-247
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• 2020

We have expressed extracellular poly(3-hydroxybutyrate) (PHB) depolymerase of Ralstonia pickettii T1 on the Escherichia coli surface using Pseudomonas OprF protein as a fusion partner by C-terminal deletion-fusion strategy. Surface display of depolymerase was confirmed by flow cytometry, immunofluorescence microscopy and whole cell hydrolase activity. For the application, depolymerase was used as an immobilized catalyst of enantioselective hydrolysis reaction for the first time. After 48 h, (R)-methyl mandelate was completely hydrolyzed, and (S)-mandelic acid was produced with over 99% enantiomeric excess. Our findings suggest that surface displayed depolymerase on E. coli can be used as an enantioselective biocatalyst.

• BMB Reports
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• 제48권10호
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• pp.583-588
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• 2015

Microtubule actin crosslinking factor 1 (MACF1), a widely expressed cytoskeletal linker, plays important roles in various cells by regulating cytoskeleton dynamics. However, its role in osteoblastic cells is not well understood. Based on our previous findings that the association of MACF1 with F-actin and microtubules in osteoblast-like cells was altered under magnetic force conditions, here, by adopting a stable MACF1-knockdown MC3T3-E1 osteoblastic cell line, we found that MACF1 knockdown induced large cells with a binuclear/multinuclear structure. Further, immunofluorescence staining showed disorganization of F-actin and microtubules in MACF1-knockdown cells. Cell counting revealed significant decrease of cell proliferation and cell cycle analysis showed an S phase cell cycle arrest in MACF1-knockdown cells. Moreover and interestingly, MACF1 knockdown showed a potential effect on cellular MTT reduction activity and mitochondrial content, suggesting an impact on cellular metabolic activity. These results together indicate an important role of MACF1 in regulating osteoblastic cell morphology and function.

• Journal of Plant Biology
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• 제29권2호
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• pp.85-94
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• 1986

Canavanine content of the cotyledons of Canavalia lineata decreased gradually during germination and growth of seedlings but continued to increase in roots and leaves. After abscission of cotyledons, canavanine content of leaves depleted competely. The activity of canavanase could be detected in leaves and roots, but not in cotyledons. High arginase activity was observed in the cotyledons of seeds at the earlyimbibition period. During the growth of seedlings, cotyledonary canavanine appeared to be transported to the growing of seedlings where it could be utilized through nitrogen metabolic pathways. In crude cell-free extracts of leaves, maximum activities of canavanase or arginase appeared in 30mM Tris-HCl buffer (pH 9.0) or 30mM NaHCO3 buffer (pH 10.0), respectively. The activities of these two enzymes differed from each other when treated with Co2+ or Mn2+. These results support the idea that canavanase and arginase might be different enzymes.

• 한국환경농학회지
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• 제16권2호
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• pp.142-148
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• 1997

중금속오염폐수처리에의 미생물 이용 가능성을 검토하기 위하여 중금속에 강한 내성을 지님과 동시에 균체내 중금속 축적능력이 우수한 중금속 내성균을 분리하여 competing ion, 대사저해제 등과 같은 외부 요인에 따른 균체내 중금속 축적변화를 조사한 결과는 다음과 같다. 중금속을 처리한 용액중 양이온인 $Al^{3+}$이온과 음이온인 $CO_3\;^{2-}$ 및 $PO_4\;^{2-}$이온이 competing ion으로 존재할 경우 중금속 내성균의 균체내 중금속 축적은 크게 감소되었으나, 그외 다른 양이온들과 음이온들에 대해서는 거의 영향을 받지 않았다. Cd, Zn 및 Cu 내성균주의 Cd, Zn 및 Cu 축적은 대사저해제에 의하여 크게 감소되었으나, Pb 내성균주에 의한 Pb 축적은 거의 영향을 받지 않는 것으로 나타나 Cd, Zn 및 Cu 내성균주의 Cd, Zn 및 Cu 축적은 에너지 의존적인 과정으로서 물질대사와 관련되어 있는 것으로 생각되었으며, Pb 내성균주의 Pb 축적은 에너지 비의존적인 과정으로서 물질대사와 관련이 별로 없는 물리적인 과정에 의하여 축적되는 것으로 생각되었다. 각 중금속 내성균주들은 해당 중금속 축적능력에 비하여 낮았으나 타 중금속들의 축적능력도 있었으며, 여러가지 중금속들을 동일 농도로 복합처리 하였을 경우 해당 중금속들의 축적능력이 타 중금속들에 비하여 매우 높게 나타나 각 중금속 내성균주들의 해당 중금속 축적은 선택적인 과정에 의하여 축적되는 것으로 생각되었다.

• 한국환경독성학회:학술대회논문집
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• 한국환경독성학회 2003년도 춘계학술대회
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• pp.185-185
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• 2003

To develope the novel anti-allergic drug, many sophoricoside derivatives were synthesized. Among these derivatives, JSH-II-3, JSH-Ⅵ-3, JSH-Ⅶ-3, and JSH-Ⅷ-3 were selected and subjected to high throughput toxicity screening (HTTS) because they revealed strong IL-5 inhibitory activity and limitation of quantity. Mouse lymphoma thymidine kinase (tk$\^$+/-/) gene assay (MOLY) and single cell gel electrophoresis (Comet) assay in mammalian cells were used as HTTS tool in our laboratory. In MOLY assay, JSH-Ⅶ-3 at 50 ∼ 6 $\mu\textrm{g}$/ml concentrations was not shown significant mutagenic effect in the absence and presence of S-9 metabolic activation system. However, the concentration of ISH-II-3, 38 $\mu\textrm{g}$/ml, induced increased mutation frequency (MF) in the presence of S-9 metabolic activation system. Also in comet assay, DNA damage was not observed in JSH-Ⅵ-3 and JSH-Ⅶ-3, wherase concentration of 32.8 $\mu\textrm{g}$/ml in JSH-II-3 and 13.9 $\mu\textrm{g}$/ml in JSH-Ⅶ-3 were induced DNA damage in the absence of S-9 metabolic activation system. Therefore, we suggest that JSH-Ⅵ-3 and JSH-Ⅶ-3 have no genotoxic effects but JSH-II-3 and JSH-Ⅷ-3 induce some mutagenicity and DNA strand breaks in mouse lymphoma cell line used this study.

• BMB Reports
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• 제54권9호
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• pp.464-469
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• 2021

Cardiomyocyte differentiation occurs through complex and finely regulated processes including cardiac lineage commitment and maturation from pluripotent stem cells (PSCs). To gain some insight into the genome-wide characteristics of cardiac lineage commitment, we performed transcriptome analysis on both mouse embryonic stem cells (mESCs) and human induced PSCs (hiPSCs) at specific stages of cardiomyocyte differentiation. Specifically, the gene expression profiles and the protein-protein interaction networks of the mESC-derived platelet-derived growth factor receptor-alpha (PDGFRα)⁺ cardiac lineage-committed cells (CLCs) and hiPSC-derived kinase insert domain receptor (KDR)⁺ and PDGFRα⁺ cardiac progenitor cells (CPCs) at cardiac lineage commitment were compared with those of mesodermal cells and differentiated cardiomyocytes. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway analyses revealed that the genes significantly upregulated at cardiac lineage commitment were associated with responses to organic substances and external stimuli, extracellular and myocardial contractile components, receptor binding, gated channel activity, PI3K-AKT signaling, and cardiac hypertrophy and dilation pathways. Protein-protein interaction network analysis revealed that the expression levels of genes that regulate cardiac maturation, heart contraction, and calcium handling showed a consistent increase during cardiac differentiation; however, the expression levels of genes that regulate cell differentiation and multicellular organism development decreased at the cardiac maturation stage following lineage commitment. Additionally, we identified for the first time the protein-protein interaction network connecting cardiac development, the immune system, and metabolism during cardiac lineage commitment in both mESC-derived PDGFRα⁺ CLCs and hiPSC-derived KDR⁺PDGFRα⁺ CPCs. These findings shed light on the regulation of cardiac lineage commitment and the pathogenesis of cardiometabolic diseases.

• Nutrition Research and Practice
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• 제18권1호
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• pp.46-61
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• 2024

BACKGROUND/OBJECTIVES: An increasing life expectancy in society has burdened healthcare systems substantially because of the rising prevalence of age-related metabolic diseases. This study compared the effects of animal protein hydrolysate (APH) and casein on metabolic diseases using aged mice. MATERIALS/METHODS: Eight-week-old and 50-week-old C57BL/6J mice were used as the non-aged (YC group) and aged controls (NC group), respectively. The aged mice were divided randomly into 3 groups (NC, low-APH [LP], and high-APH [HP] and fed each experimental diet for 12 weeks. In the LP and HP groups, casein in the AIN-93G diet was substituted with 16 kcal% and 24 kcal% APH, respectively. The mice were sacrificed when they were 63-week-old, and plasma and hepatic lipid, white adipose tissue weight, hepatic glucose, lipid, and antioxidant enzyme activities, immunohistochemistry staining, and mRNA expression related to the glucose metabolism on liver and muscle were analyzed. RESULTS: Supplementation of APH in aging mice resulted in a significant decrease in visceral fat (epididymal, perirenal, retroperitoneal, and mesenteric fat) compared to the negative control (NC) group. The intraperitoneal glucose tolerance test and area under the curve analysis revealed insulin resistance in the NC group, which was alleviated by APH supplementation. APH supplementation reduced hepatic gluconeogenesis and increased glucose utilization in the liver and muscle. Furthermore, APH supplementation improved hepatic steatosis by reducing the hepatic fatty acid and phosphatidate phosphatase activity while increasing the hepatic carnitine palmitoyltransferase activity. Furthermore, in the APH supplementation groups, the red blood cell (RBC) thiobarbituric acid reactive substances and hepatic H₂O₂ levels decreased, and the RBC glutathione, hepatic catalase, and glutathione peroxidase activities increased. CONCLUSIONS: APH supplementation reduced visceral fat accumulation and alleviated obesity-related metabolic diseases, including insulin resistance and hepatic steatosis, in aged mice. Therefore, high-quality animal protein APH that reduces the molecular weight and enhances the protein digestibility-corrected amino acid score has potential as a dietary supplement for healthy aging.

• The Korean Journal of Physiology and Pharmacology
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• 제19권5호
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• pp.389-399
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• 2015

Zinc has been considered as a vital constituent of proteins, including enzymes. Mobile reactive zinc ($Zn^{2+}$) is the key form of zinc involved in signal transductions, which are mainly driven by its binding to proteins or the release of zinc from proteins, possibly via a redox switch. There has been growing evidence of zinc's critical role in cell signaling, due to its flexible coordination geometry and rapid shifts in protein conformation to perform biological reactions. The importance and complexity of $Zn^{2+}$ activity has been presumed to parallel the degree of calcium's participation in cellular processes. Whole body and cellular $Zn^{2+}$ levels are largely regulated by metallothioneins (MTs), $Zn^{2+}$ importers (ZIPs), and $Zn^{2+}$ transporters (ZnTs). Numerous proteins involved in signaling pathways, mitochondrial metabolism, and ion channels that play a pivotal role in controlling cardiac contractility are common targets of $Zn^{2+}$. However, these regulatory actions of $Zn^{2+}$ are not limited to the function of the heart, but also extend to numerous other organ systems, such as the central nervous system, immune system, cardiovascular tissue, and secretory glands, such as the pancreas, prostate, and mammary glands. In this review, the regulation of cellular $Zn^{2+}$ levels, $Zn^{2+}$-mediated signal transduction, impacts of $Zn^{2+}$ on ion channels and mitochondrial metabolism, and finally, the implications of $Zn^{2+}$ in health and disease development were outlined to help widen the current understanding of the versatile and complex roles of $Zn^{2+}$.

• Applied Microscopy
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• 제22권1호
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• pp.15-32
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• 1992

This study was an attempt to investigate and compare the fine structure of small cells in the space of Disse of various animal livers. All animal livers contained small cells with or without lipid droplets and the one with lipid droplet seemed to be more developed and show an abundance of activity in function. The fine structure of the small cells observed in the nonmammals was similar to that of Ito cell in the mammal. The electron density of the small cells was similar to that of other cell types in the same animal liver. The cisternal dilation of rough endoplasmic reticulum and Golgi apparatus was more predominant in the mammalian Ito cells. In the nonmammalian, aquatic vertebrates, however, lysosomes and filaments are much more abundant in the Ito cell and its abundant cytoplasmic processes rich in filaments were usually extended between the parenchymal cells. The disparity in size of organelles and numbers of lipid droplets in the small cells showed a tendency similar to those of other cell types in the same animal. From these results, it is considered that the small cells in the space of Disse is a Ito cell and the Ito cell without lipid droplets differentiates into the one containing lipid droplets according to the characteristics of the different animals respectively, and that the Ito cells in the mammals are more active in metabolic function, while those in the nonmammalian aquatic vertebrates are abundant in support of parenchyme.

• 한국동물학회지
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• 제16권2호
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• pp.109-118
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• 1973

동면전, 중, 후 및 활동기 동안 조선산개구리(Rana amurensis coreana Okada)의 위장관 점막내에 출현하는 enterochromaffin cell의 내분비성 분비과립의 변화를 형태학적으로 관찰하여 다음과 같은 결과를 얻었다. 1. 활동기 및 동면전, 중, 후기의 위장관 점막상피 내 enterochromaffin cell의 출현비율은 1: 1.5 :0.36 : 0.9로서 본 세포의 수는 활동기군에 비하여 동면전기군은 유의하게 증가하였으며 동면 중기군은 유의하게 감소하였다. 2. 활동기 및 동면전기군의 EC cell은 대체로 그 세포질내에 분비과립이 차있는 다과립성 및 중자등과립성 세포가 대다수였으나 동면중기 및 후기군에서는 희과립세포가 현저히 증가되었다. 3. EC cell의 형태학적 변화는 다른 신진대사 특히 내분비적 활성의 약화로 야기되는 생체의 적응현상에서 온 결과라고 추측된다.