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Identification and Functional Analysis of Pig β-1,4-N-Acetylglucosaminyltransferase A (MGAT4A) (돼지 유래의 β-1,4-N-acetylglucosaminyltransferase A (MGAT4A) 유전자의 동정 및 기능 분석)

  • Kim, Ji-Youn;Hwang, Hwan-Jin;Chung, Hak-Jae;Park, Mi-Ryung;Byun, Sung June;Kim, Kyung-Woon
    • Journal of Life Science
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    • v.26 no.3
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    • pp.275-281
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    • 2016
  • Glycan modification is important in pharmaceutical industry. Especially, sialic acid affects the bioactivity and stability of medicine. Milk of pig has been used as bioreactor to produce various pharmaceutical proteins. Therefore, it is necessary to modify the glycan chain in pig mammary grand. β-1,4-N-Acetylglucosaminyltransferase A (pMGAT4A) is one of the essential enzymes for increase of sialic acid content, but pig MGAT4A is unclear. In this study, the pMGAT4A was identified and characterized. The pMGAT4A has 1638 nucleotides encoding 535 amino acids and type II membrane topology, which is one of the common features in many glycosyltransferases. The gene was strongly expressed in liver and mammary gland, whereas was weakly expressed in small intestine, stomach and bladder. For functional test, HA-tagged MGAT4A was over-expressed in porcine kidney (PK-15) cell line. Forced expression of pMGAT4A gene was identified by qPCR, and we identified that pMGAT4A is located in Golgi complex by co- staining with HA antibody and BODIPY TR ceramide. In addition, we identified the increase of mannose-β-1,4-N-acetylglucosamine structure by ELISA and immunofluorescence using Datura stramonium agglutinin (DSA), which recognizes mannose-β-1,4-Nacetylglucosamine. Through the specific activity analysis, we showed that pMGAT4A modified bi-antennary to tri-antennary. This event affects sialic acid content. Therefore, we thought that over-expression of pMGAT4A will be necessary in pig mammary grand for improved medicine.

Isolation and Characterization of Bacillus subtilis MP56 with Antimicrobial Activity against MDR (Multi Drug Resistant) Strains (다약제내성균에 대한 항균 활성을 가지는 Bacillus subtilis MP56 균주의 분리 및 특성분석)

  • Park, Sungyong;Yoo, Jincheol;Seong, Chinam;Cho, Seungsik
    • Korean Journal of Microbiology
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    • v.49 no.1
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    • pp.90-94
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    • 2013
  • A new Bacillus strain designated as MP56 producing antimicrobial substance has been isolated from the mud flat of Korea. The strain MP56 was found to exhibit broad spectrum of antimicrobial activity against Gram-positive pathogenic microorganisms and MDR (multi drug resistant) strains. The 16S rRNA sequence revealed that the MP56 was closely related to Bacillus subtilis with 99.93% homology. The optimal medium composition for production of antimicrobial substance in the B. subtilis MP56 were 1% mannitol, 1% oat meal, 0.01% $CaCl_2$. Antimicrobial activity of the culture broth against different pathogenic strains was assessed using the antimicrobial spectrum. The result suggests that Bacillus strain MP56 produces high quality antimicrobial substance that might be very useful to control varieties of pathogenic microbial growth.

Sulfasalazine Induces Apoptosis and Cell Cycle Arrest in RAW 264.7 Macrophages (마우스 대식세포에서 설파살라진의 세포사멸 및 세포주기 정체에 미치는 영향 연구)

  • Seong Mi Kim;Sohyeon Park ;Jin-Kyung Kim
    • Journal of Life Science
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    • v.33 no.10
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    • pp.767-775
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    • 2023
  • Sulfasalazine is a disease-modifying antirheumatic abiotic agent. It is a derivative of aminosalicylic acid and has been used for the treatment of various inflammatory diseases, such as rheumatoid arthritis, ulcerative colitis, and Crohn's disease, since it was first synthesized in 1941 and approved as a medicine in the United States in 1950. However, its mechanism of action has not yet been clearly identified. In this study, the effects of sulfasalazine on cell survival, apoptosis, and cell cycle progression in macrophages, which are major immune cells that regulate inflammatory responses, were investigated using mouse macrophage RAW 264.7 cells. Sulfasalazine inhibited the viability of RAW 264.7 cells in a dose-dependent manner, starting at a concentration of 0.25 mM. Annexin-V staining was used to confirm that the decrease in cell viability was due to apoptosis, and the number of Annexin-V-positive cells increased significantly at a concentration of 0.25 mM or higher. The effect of sulfasalazine on the expression of key proteins that regulate the G0/G1 phase of the cell cycle was also investigated. Sulfasalazine treatment significantly increased the expression of the cyclin-dependent kinase inhibitors p21 and p27 in RAW 264.7 cells. Although sulfasalazine is frequently used as a control drug in studies on inflammatory diseases, such as inflammatory colitis and rheumatoid arthritis, studies on its effect on macrophages are very limited. Therefore, the results of this study are expected to provide vital information on the use of sulfasalazine as a disease treatment.

Effects of Radiation Mutant Perilla frutescens var. crispa in Preventing and Alleviating Symptoms in a Monosodium Iodoacetate-Induced Osteoarthritis Rat Model (MIA로 퇴행성관절염을 유도한 랫드에 방사선 형질전환 차조기가 증상 예방 및 완화에 미치는 효과)

  • Sim, Boo-Yong;Joo, In-Hwan;Kim, Sung-Kyu;Ji, Joong-Gu
    • Journal of the Korean Applied Science and Technology
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    • v.37 no.4
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    • pp.830-838
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    • 2020
  • The present study aimed to evaluate the effect of radiation mutant Perilla frutescens var. crispa on bone metabolism and the inflammatory response in a monosodium iodoacetateinduced rat model of osteoarthritis. radiation mutant Perilla frutescens var. crispa was administered orally at doses of 25, 50 or 100 mg/kg/day for 2 weeks before direct injection of monosodium iodoacetate (3 mg/50 μl of 0.9% saline) into the intra-articular space of the rats' right knees. The rats subsequently received the same doses of oral radiation mutant Perilla frutescens var. crispa for another 4 weeks. It was evaluated that the treatment effects based on serum biomarkers, and morphological and histopathological analysis of the knee joints. Compared with those in control rats, the radiation mutant Perilla frutescens var. crispa treatments significantly reduced the serum levels of inflammation, bone metabolism markers (i.e., COX-2, LTB4, MMP-3, and COMP), and the amount of fibrous tissue. Otherwise, it was significantly increased the concentration of TIMP-1 and calcitonin. In addition, the radiation mutant Perilla frutescens var. crispa treatments effectively preserved the knee cartilage and synovial membrane. As a result, it indicates that radiation mutant Perilla frutescens var. crispa prevented and alleviated osteoarthritis symptoms. Thus, radiation mutant Perilla frutescens var. crispa can be used in food and drug material for the management of osteoarthritis.

Pharmacokinetics and the Intestinal Permeability of Amaranth in Rats (적색식용색소인 아마란스의 약동학 특성 및 위장관 투과도 연구)

  • Han, Youjin;Goo, Soo Hyeon;Nam, So Jeong;Kang, Yun Ju;Kwon, Mihwa;Song, Im-Sook
    • Journal of Life Science
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    • v.27 no.7
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    • pp.812-816
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    • 2017
  • Although amaranth, a red-colored tar dye, is usually used in foods, cosmetics, and pharmaceutics, its bioavailability and intestinal absorption have not previously been investigated. Therefore, the purpose of this study was to investigate the pharmacokinetics properties and intestinal permeability of amaranth in rats following the intravenous and oral administration of this dye. Amaranth rapidly disappeared from the plasma following the intravenous injection, with a half-life of 38.8 minutes. However, the plasma concentration of amaranth was increased to 400 minutes following the oral administration of amaranth, and the absorption time and bioavailability of amaranth were calculated to be 356 minutes and 55.6%, respectively. This suggests that once amaranth exists in the gut, this dye may be efficiently and effectively absorbed. Consistent with this result, the intestinal permeability of amaranth was comparable to atenolol, a marker compound of moderate permeability, and to one-third of caffeine's intestinal permeability (a highly permeable compound). In conclusion, a significantly long absorption time and substantial intestinal absorption of amaranth was observed following the oral administration of amaranth at a dose of 300 mg/kg in rats, despite the rapid elimination of this dye from the plasma. These results may suggest the necessity of a careful and limited use of amaranth dye when it is added to food, lip-care cosmetics, and orally administered pharmaceutics.

Use of Fraxnius rhynchophylla H.ance bark as antioxidant (물푸레나무 수피의 항산화 효과)

  • 김대구;이병근;김현용
    • Journal of Korea Foresty Energy
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    • v.22 no.2
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    • pp.69-77
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    • 2003
  • This study reviewed antioxidants and various enzymic physiological functions of Korean ash tree extractives that contains coumarine and ester component in order to examine the practical possibility of those materials as pharmaceutical and cosmetic additive applications. first, in the experiment of enzymic physiological activity, the electron donating ability showed 62.00% at 1ppm and over 90% at 10ppm or higher concentration, and the pseudo SOD ability showed 39% at the concentration of 2000ppm and 84.73% at the concentration of 5000ppm. Second, the inhibition activity of xanthine oxidase showed a relatively low inhibition effect of 45.57% at 200ppm, and the inhibition effect of tyrosinase showed 57.51% at 2000ppm. However, it showed a very low inhibition effect at the concentration of 500ppm or less. Third, the lipid oxidation, when Cu$\^$2+/ion was used as an oxidation accelerator, showed the inhibition effect of 82% at the concentration of 10ppm, and when Fe$\^$2+/ion was used also as the oxidation accelerator, it showed a very outstanding effect of 60% and more at 100ppm. Conclusively it is clear that the extracts of Korean ash tree bark can be used as cosmetic and pharmaceutical additive agents due to their excellent activities as antioxidant and enzymic physiological functions of it.

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Anti-inflammatory and Anti-Atopic Effects of Crude Extracts and Solvent Fractions of Phormium tenax leaf (신서란(Phormium tenax) 잎 조추출물 및 용매 분획물의 항염증 및 항아토피 효과)

  • Yang, Kwon Min;Song, Sang mok;Lee, Doseung;Yoon, Weon-Jong;Kim, Chan-Shick;Kim, Chang Sook
    • Korean Journal of Plant Resources
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    • v.32 no.5
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    • pp.433-441
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    • 2019
  • This study describes a preliminary evaluation of the anti-inflammatory activity and anti-atopic activity of Phormium tenax leaf extracts. P. tenax leaf was extracted using 70% ethanol and then fractionated sequentially with n-hexane, methylene chloride, ethyl acetate, n-butanol. In order to effectively screen for anti-inflammatory agents, we first investigated the inhibitory effects of P. tenax leaf crude extracts and solvent fractions on production of pro-inflammatory factors[nitric oxide(NO), prostaglandin $E_2(PGE_2)$, inducible nitric oxide synthase(iNOS) and cyclooxygenase-2(COX-2)] and pro-inflammatory cytokines [tumor necrosis $factor-{\alpha}(TNF-{\alpha})$, interleukin-6(IL-6) and $interleukin-1{\beta}(IL-1{\beta})$] in lipopolysaccharide(LPS)-stimulated RAW 264.7 cells. In addition, we also evaluated of their inhibitory effect on the atopic dermatitis-like inflammatory markers such as macrophage-derived chemokine(MDC) and thymus and activation-regulated chemokine(TARC) in HaCaT cells. Among the five solvent fractions of P. tenax, methylene chloride and ethyl acetate fractions inhibited production of pro-inflammatory factors and pro-inflammatory cytokines in a dose dependent manner, respectively. These fractions were also showed inhibitory activity for MDC and TARC expression levels in $IFN-{\gamma}-stimulated$ HaCaT cells, respectively. These results suggest that P. tenax have significantly effects of anti-inflammatory activity and anti-atopic activity that might be beneficial for the topical treatment of inflammatory skin disorders.

Application of Molecular Diagnostics Technology in the Development of a Companion Diagnostics for Malignant Solid Tumors (악성 고형암의 항암제 동반진단 기술에서 분자진단기술의 적용)

  • Kim, Jin-Hee
    • The Journal of the Korea Contents Association
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    • v.19 no.3
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    • pp.365-374
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    • 2019
  • Unlike benign tumors, malignant tumors are capable of metastasis, easy to relapse, poor survival, and low quality of life. In Korea, here is a tendency to treat the tumors collectively according to the General Principles of Cancer Chemotherapy(GPCC) of the Health Insurance Review & Assessment Service (HIRA). But recently, companion diagnostics(CDx) is recommended rather than unilateral medication because biomarker-based molecular diagnostics is possible to predict the drug response of patients before drug treatment. Not only domestic but also overseas Food and Drug Administratio (FDA) recommends the development of the CDx system at the stage of drug development to ensure the responsiveness and safety of medicines. In this study, I focused on the necessity of CDx development direction as well as CDx development status through literature review. Furthermore I also discussed CDx types according to the molecular diagnostic technology such as immunohistochemistry (IHC), polymerase chain reaction (PCR), in situ hybridization (ISH), and next-generation sequencing (NGS) not only in the approved CDx but also in the developing one by US FDA. And I suggested the technology issue of CDx development process such as a selection of molecular diagnostics at the time of release, a clear understanding of the CDx mechanism, and a convergence of drug with CDx development. The necessity of social insurance system also was proposed for CDx development.

Inhibitory Effects of Four Solvent Fractions of Alnus firma on α-Amylase and α-Glucosidase. (사방오리나무 추출물의 α-amylase 및 α-glucosidase 저해활성)

  • Choi, Hye-Jung;Jeong, Yong-Kee;Kang, Dae-Ook;Joo, Woo-Hong
    • Journal of Life Science
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    • v.18 no.7
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    • pp.1005-1010
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    • 2008
  • In this study, we investigated the inhibitory effect of four solvent fractions of Alnus firma on ${\alpha}-amylase$, ${\alpha}-glucosidase$ and aldose reductase activities. The inhibitory test showed that methanol (MeOH) extract and hexane (HX) fraction strongly inhibited pork pancreatin and salivary ${\alpha}-amylase$ activity. The MeOH extract and HX fraction of Alnus firma at the concentration of 4 mg/ml inhibited more than 70% of pancreatin and salivary ${\alpha}-amylase$ activity. The inhibitory effect of fractions has different specificities against ${\alpha}-amylase$ from pancreatin and salivary. In addition, the MeOH extract and butanol (BuOH) fraction showed the highest inhibitory activity on yeast ${\alpha}-glucosidase$ at values of $IC_{50}$ $137.36\;{\mu}g/ml$ and $115.14\;{\mu}g/ml$ respectively. The MeOH extract and BuOH fraction showed the highest inhibitory activity on yeast ${\alpha}-glucosidase$ than commercial agent such as 1-deoxynorjirimycin and acarbose. Inhibition kinetics of solvent fractions showed that ${\alpha}-glucosidase$ has been inhibited noncompetitively by the MeOH, EA and BuOH fraction. The aldose reductase from human muscle cell had been inhibited strongly by the MeOH extract and EA fraction at 57.996% and 83.293% at the concentration of $50\;{\mu}g/ml$, respectively. These findings may contribute to biological significance in that ${\alpha}-amylase$, ${\alpha}-glucosidase$ and aldose reductase inhibitory compounds could be used as a functional food and a drug for the symptomatic treatment of antidiabetic disease in the future.

Single Cell Oil-Recent Trends in Microbial Production and Utilization (미생물에 의한 단세포유지의 생산과 이용)

  • Kim, Yong-Ro;Yoon, Suk Hoo
    • Korean Journal of Food Science and Technology
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    • v.47 no.6
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    • pp.687-697
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    • 2015
  • With the shortage of edible fats and oils and depletion of fossil fuels in many countries, microbial lipids is emerging as one of the most promising sources of fats and oils in the global market. Oleaginous microorganisms, also called single cell oils (SCOs), can accumulate lipids more than 25% in the cell volume. Triacylglycerols are the major storage lipids. SCOs offer several advantages for lipid production as follows: SCOs have short life span which would shorten production time, cultivation conditions are not affected by climate and place; the production process is easy to control. There are a number of oleaginous yeasts, molds, and microalgae. Furthermore, the lipid productivity of SCOs can be enhanced through strain improvement and the optimization of cultivation conditions. The new strains developed using recent advanced biotechnical methods showed greatly improved oleaginicity. Further, hydrolysates of lignocellulosic materials can be used as carbon sources for economic production of SCO.