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Acceleration of Mesenchymal-to-Epithelial Transition (MET) during Direct Reprogramming Using Natural Compounds

  • Seo, Ji-Hye;Jang, Si Won;Jeon, Young-Joo;Eun, So Young;Hong, Yean Ju;Do, Jeong Tae;Chae, Jung-il;Choi, Hyun Woo
    • Journal of Microbiology and Biotechnology
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    • v.32 no.10
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    • pp.1245-1252
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    • 2022
  • Induced pluripotent stem cells (iPSCs) can be generated from somatic cells using Oct4, Sox2, Klf4, and c-Myc (OSKM). Small molecules can enhance reprogramming. Licochalcone D (LCD), a flavonoid compound present mainly in the roots of Glycyrrhiza inflata, acts on known signaling pathways involved in transcriptional activity and signal transduction, including the PGC1-α and MAPK families. In this study, we demonstrated that LCD improved reprogramming efficiency. LCD-treated iPSCs (LCD-iPSCs) expressed pluripotency-related genes Oct4, Sox2, Nanog, and Prdm14. Moreover, LCD-iPSCs differentiated into all three germ layers in vitro and formed chimeras. The mesenchymal-to-epithelial transition (MET) is critical for somatic cell reprogramming. We found that the expression levels of mesenchymal genes (Snail2 and Twist) decreased and those of epithelial genes (DSP, Cldn3, Crb3, and Ocln) dramatically increased in OR-MEF (OG2+/+/ROSA26+/+) cells treated with LCD for 3 days, indicating that MET effectively occurred in LCD-treated OR-MEF cells. Thus, LCD enhanced the generation of iPSCs from somatic cells by promoting MET at the early stages of reprogramming.

Endoplasmin regulates differentiation of tonsil-derived mesenchymal stem cells into chondrocytes through ERK signaling

  • Kim, Hye Ryeong;Choi, Hyeongrok;Park, Soon Yong;Song, Young-Chul;Kim, Jae-Ho;Shim, Sangin;Jun, Woojin;Kim, Kyung-Jin;Han, Jin;Chi, Seung-Wook;Leem, Sun-Hee;Chung, Jin Woong
    • BMB Reports
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    • v.55 no.5
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    • pp.226-231
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    • 2022
  • It is well-known that some species of lizard have an exceptional ability known as caudal autotomy (voluntary self-amputation of the tail) as an anti-predation mechanism. After amputation occurs, they can regenerate their new tails in a few days. The new tail section is generally shorter than the original one and is composed of cartilage rather than vertebrae bone. In addition, the skin of the regenerated tail distinctly differs from its original appearance. We performed a proteomics analysis for extracts derived from regenerating lizard tail tissues after amputation and found that endoplasmin (ENPL) was the main factor among proteins up-regulated in expression during regeneration. Thus, we performed further experiments to determine whether ENPL could induce chondrogenesis of tonsil-derived mesenchymal stem cells (T-MSCs). In this study, we found that chondrogenic differentiation was associated with an increase of ENPL expression by ER stress. We also found that ENPL was involved in chondrogenic differentiation of T-MSCs by suppressing extracellular signal-regulated kinase (ERK) phosphorylation.

Anti-inflammatory and antioxidant activities of Sargassum horneri extract in RAW264.7 macrophages

  • Kim, Min Ju;Jo, Hee Geun;Ramakrishna, Chilakala;Lee, Seung-Jae;Lee, Dong-Sung;Cheong, Sun Hee
    • Korean Journal of Exercise Nutrition
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    • v.25 no.4
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    • pp.45-53
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    • 2021
  • [Purpose] In this study, we investigated whether a 70% ethanolic (EtOH) extract of Sargassum horneri had antioxidant and anti-inflammatory effects in lipopolysaccharide (LPS)-stimulated macrophage-like RAW 264.7 cells. [Methods] The proximate composition, fatty acids, amino acids, and dietary fiber of S. horneri, various biologically active compounds, and antioxidant activity were analyzed. [Results] The DPPH and ABTS free radical scavenging activities, as well as the reduction power, of the S. horneri extract used here were significantly increased in a concentration-dependent manner. This indicates that S. horneri contains bioactive compounds, such as phenols and flavonoids, that have excellent antioxidant activity. The cellular viability and metabolic activity results confirmed that the extract had no discernible toxicity at concentrations up to 100 ㎍/mL. The levels of nitrites and cytokines (PGE2, TNF-α and IL-6), which mediate pro-inflammatory effect, were significantly inhibited by treatment with either 50 or 100 ㎍/mL S. horneri extract, whereas that of IL-1β was significantly inhibited by treatment with 100 ㎍/mL of the extract. Similarly, the expression of iNOS and COX-2 proteins also decreased according to 50 or 100 ㎍/mL extract concentrations. NF-κB binding to DNA was also significantly inhibited by treatment with 100 ㎍/mL of extract. [Conclusion] These results suggest that 70% EtOH extracts of S. horneri can relieve inflammation caused by disease or high intensity exercise.

Structural resemblance of the DNAJA-family protein, Tid1, to the DNAJB-family Hsp40

  • Jang, Jinhwa;Lee, Sung-Hee;Kang, Dong-Hoon;Sim, Dae-Won;Ryu, Kyung-Suk;Jo, Ku-Sung;Lee, Jinhyuk;Ryu, Hyojung;Kim, Eun-Hee;Won, Hyung-Sik;Kim, Ji-Hun
    • BMB Reports
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    • v.55 no.10
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    • pp.488-493
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    • 2022
  • The specific pair of heat shock protein 70 (Hsp70) and Hsp40 constitutes an essential molecular chaperone system involved in numerous cellular processes, including the proper folding/refolding and transport of proteins. Hsp40 family members are characterized by the presence of a conserved J-domain (JD) that functions as a co-chaperone of Hsp70. Tumorous imaginal disc 1 (Tid1) is a tumor suppressor protein belonging to the DNAJA3 subfamily of Hsp40 and functions as a co-chaperone of the mitochondrial Hsp70, mortalin. In this work, we performed nuclear magnetic resonance spectroscopy to determine the solution structure of JD and its interaction with the glycine/phenylalanine-rich region (GF-motif) of human Tid1. Notably, Tid1-JD, whose conformation was consistent with that of the DNAJB1 JD, appeared to stably interact with its subsequent GF-motif region. Collectively with our sequence analysis, the present results demonstrate that the functional and regulatory mode of Tid1 resembles that of the DNAJB1 subfamily members rather than DNAJA1 or DNAJA2 subfamily proteins. Therefore, it is suggested that an allosteric interaction between mortalin and Tid1 is involved in the mitochondrial Hsp70/Hsp40 chaperone system.

The roles of growth factors and hormones in the regulation of muscle satellite cells for cultured meat production

  • Syed Sayeed Ahmad;Hee Jin Chun;Khurshid Ahmad;Sibhghatulla Shaikh;Jeong Ho Lim;Shahid Ali;Sung Soo Han;Sun Jin Hur;Jung Hoon Sohn;Eun Ju Lee;Inho Choi
    • Journal of Animal Science and Technology
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    • v.65 no.1
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    • pp.16-31
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    • 2023
  • Cultured meat is a potential sustainable food generated by the in vitro myogenesis of muscle satellite (stem) cells (MSCs). The self-renewal and differentiation properties of MSCs are of primary interest for cultured meat production. MSC proliferation and differentiation are influenced by a variety of growth factors such as insulin-like growth factors (IGF-1 and IGF-2), transforming growth factor beta (TGF-β), fibroblast growth factors (FGF-2 and FGF-21), platelet-derived growth factor (PDGF) and hepatocyte growth factor (HGF) and by hormones like insulin, testosterone, glucocorticoids, and thyroid hormones. In this review, we investigated the roles of growth factors and hormones during cultured meat production because these factors provide signals for MSC growth and structural stability. The aim of this article is to provide the important idea about different growth factors such as FGF (enhance the cell proliferation and differentiation), IGF-1 (increase the number of myoblasts), PDGF (myoblast proliferation), TGF-β1 (muscle repair) and hormones such as insulin (cell survival and growth), testosterone (muscle fiber size), dexamethasone (myoblast proliferation and differentiation), and thyroid hormones (amount and diameter of muscle fibers and determine the usual pattern of fiber distributions) as media components during myogenesis for cultured meat production.

Physicochemical properties and physiological activity of bean sprouts extract containing Hovenia dulcis Thunb concentrates

  • Ji-An Heo;Wool-Lim Park;Hye-Ji Min;Jeong-Ho Kim;Yeong-Seon Won;Kwon-Il Seo
    • Food Science and Preservation
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    • v.30 no.4
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    • pp.617-629
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    • 2023
  • Reactive oxygen species are the byproducts of metabolic processes in the body, However, excessive amount may cause side effects such as cancer. Therefore, to reduce the production of these species, but their long-term administration at high doses may induce side effects. Hence, natural materials with antioxidant activities are attracting attention. Two of these natural materials are soybean sprouts and Hovenia dulcis Thunb. fruits, but few studies have evaluated the effects of their combination. Thus, we prepared a soybean sprout extract containing 1.5% H. dulcis Thunb. fruit concentrate (BHM) to develop a functional food material derived from natural products and then confirmed its physicochemical properties and physiological activity. Among the organic acids detected in BHM, malic acid exhibited the highest content of 1,451.03 ppm, and the main free sugars were glucose (645.48 ppm) and fructose (738.11 ppm). Taurine was the most abundant free amino acid at a concentration of 11.95 ppm, followed by those of arginine (10.97 ppm) and glutamic acid (10.16 ppm). Analyses of the mineral components revealed large amounts of Zn and Fe in BHM, and the respective total polyphenol and flavonoid contents in BHM were 957.16 and 601.93 ppm. The DPPH radical and H2O2 scavenging activities and reducing power indicated excellent antioxidant efficacy compared to the positive controls. Furthermore, blood alcohol and acetaldehyde concentrations were measured to confirm the hangover-relieving effects of BHM, with both significantly decreased (p<0.05). BHM displays potential for development as a functional food, and the results of this study may be used as basic data in further research.

Effects of Environmental Factors on the Eye Direction in Juvenile Starry Flounder Platichthys stellatus (강도다리(Platichthys stellatus) 변태 과정에서 안구 이동의 방향에 관여하는 환경적 요인)

  • Tae Min Kim;Hyun Seok Jang;Jung Yeol Park;Hyo bin Lee;Han Kyu Lim
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.57 no.4
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    • pp.448-458
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    • 2024
  • The direction of eye movement in Pleuronectiformes is species-specific. In farmed flounder, deviations in eye direction can reduce marketability. Therefore, we explored the effects of genetic and environmental factors on the direction of eye movements in the starry flounder Platichthys stellatus to address this economic challenge. Four pairs of female and male starry flounder broodstock, with eyes positioned to the right or left, were used in a mating experiment. The experimental groups were established with breeding seawater temperatures set at 10, 14, and 18℃ for the water temperature experiment. Whole-body samples of juveniles were collected to analyze the expression of genes related to eye movement and direction. The mating experiment results showed no significant differences in abnormal eye movement. In the water temperature experiment, the rate of abnormal eye movement was significantly higher at 32.19±1.33% in the 18℃ group than in the 10 and 14℃ groups. Genetic analysis of eye movement related to water temperature revealed that the expression of eye migration genes was significantly higher at 10℃. Moreover, no significant differences were observed in mating experiments. In conclusion, water temperature and not mating affected the eye movement of starry flounder.

Antifungal Activity of Bacillus sp. AM-651 Against Phytophthora capsici (고추역병 유발병원균 Phytophthora capsici에 대한 Bacillus sp. AM-651의 항진균활성)

  • Lee, Jung-Bok;Shin, Jeong-Hak;Jang, Jong-Ok;Shin, Kee-Sun;Choi, Chung-Sik;Kim, Kun-Woo;Jo, Min-Sub;Jeon, Chun-Pyo;Kim, Yun-Hoi;Kwon, Gi-Seok
    • Microbiology and Biotechnology Letters
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    • v.36 no.3
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    • pp.227-232
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    • 2008
  • Biological antagonists of Phytophthora capsici were isolated from soil in Gyeongbuk, Korea. Among the isolated bacteria, a Bacillus sp. was identified from l6S rDNA sequence analysis and named Bacillus sp. AM-651. Bacillus sp. AM-65l strain which can strongly a antifungal activity against Phytophthora capsici. Culture conditions for the maximum production of the antagonistic substance were optimized. The production of antibiotic were high on modified Davis mineral medium pH 7 at $30^{\circ}C$. The medium for highest production of the agonistic substance optimized. It is composed the best activity on glucose, $(NH_4)_2SO_4$ and $K_2HPO_4$ at 0.5%, 0.1%, and 0.7%, respectively. By time course of culture solution selected Bacillus sp. AM-65l, the culture solution after 48hrs had strongly growth inhibition rate against P. capsici. And culture solution of Bacillus sp. AM-651 was stable within a pH range $5{\sim}11$ and temperature range $4{\sim}70^{\circ}C$. Bacillus sp. AM-651 cultured broth shown fungal growth inhibitory activity against B. sorokiniana, B. cinerea, R. solani avove and beyond P. capsici and comparatively showed a high activity against C. gloeosporioides, B. dothidea, B. cinerea and F. graminearum by agar diffusion method.

Current status of sweetpotato genomics research (고구마 유전체 연구현황 및 전망)

  • Yoon, Ung-Han;Jeong, Jae Cheol;Kwak, Sang-Soo;Yang, Jung-Wook;Kim, Tae-Ho;Lee, Hyeong-Un;Nam, Sang-Sik;Hahn, Jang-Ho
    • Journal of Plant Biotechnology
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    • v.42 no.3
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    • pp.161-167
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    • 2015
  • Sweetpotato [Ipomoea batatas (L.) Lam] grows well in harsh environmental conditions, and is cultivated as one of the top seven food crops in the world. Recently, sweetpotato is drawing interest from people as a healthy food because it is high in dietary fiber, vitamins, carotenoids and overall nutrition value. However, few studies have been conducted on sweetpotato genome sequencing in spite of its importance. This review is aimed at increasing the efficiency of sweetpotato genome sequencing research as well as establishing a base for gene utilization in order to control useful traits. Recently, animal and plant genome sequencing projects increased significantly. However, sweetpotato genome sequencing has not been performed due to polyploidy and heterogeneity problems in its genome. Meanwhile research on its transcriptome has been conducted actively. Recently, a draft of the diploid sweetpotato genome was reported in 2015 by Japanese researchers. In addition, the Korea-China-Japan Trilateral Research Association of Sweetpotato (TRAS) has conducted research on gene map construction and genome sequencing of the hexaploid sweetpotato Xushu 18 since 2014. The Bill & Melinda Gates Foundation launched the 'sweetpotato genomic sequencing to develop genomic tools for Sub-Sahara Africa breeding program'. The chloroplast genome sequence acquired during sweetpotato genome sequencing is used in evolutionary analyses. In this review, the trend of research in the sweetpotato genome sequencing was analyzed. Research trend analysis like this will provide researchers working toward sweetpotato productivity and nutrient improvement with information on the status of sweetpotato genome research. This will contribute to solving world food, energy and environmental problems.

Multiple Monoclonal Antibodies Produced in a Single Transgenic Plant (형질전환 식물체에서의 복합 단일 항체 단백질 생산)

  • Ahn, Mi-Hyun;Oh, Eun-Yi;Song, Mi-Ra;Lu, Zhe;Kim, Hyun-Soon;Joung, Hyouk;Ko, Ki-Sung
    • Journal of Life Science
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    • v.19 no.1
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    • pp.123-128
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    • 2009
  • Production of highly valuable immunotherapeutic proteins such as monoclonal antibodies and vaccines using plant biotechnology and genetic engineering has been studied as a popular research field. Plant expression system for mass production of such useful recombinant therapeutic proteins has several advantages over other existing expression systems with economical and safety issues. Immunotherapy of multiple monoclonal antibodies, which can recognize multiple targeting including specific proteins and their glycans highly expressed on the surface of cancer cells, can be an efficient treatment compared to a single targeting immunotherapy using a single antibody. In this study, we have established plant production system to express two different targeting monoclonal antibodies in a single transgenic plant through crossing fertilization between two different transgenic plants expressing anti-colorectal cancer mAbCO17-1A and anti-breast cancer mAbBR55, respectively. The F1 seedlings were obtained cross fertilization between the two transgenic parental plants. The presence, transcription, and protein expression of heavy chain (HC) and light chain (LC) genes of both mAbs in the seedlings were investigated by PCR, RT-PCR, and immunoblot analyses, respectively. Among all the seedlings, some seedlings did not carry or transcribe the HC and LC genes of both mAbs. Thus, the seedlings with presence and transcription of HC and LC genes of both mAbs were selected, and the selected seedlings were confirmed to have relatively stronger density of HC and LC protein bands compared to the transgenic plant expressing only each mAb. These results indicate that the F1 seedling plant with carrying both mAb genes was established. Taken together, plant crossing fertilization can be applied to generate an efficient production system expressing multiple monoclonal antibodies for immunotherapy in a single plant.