• Title/Summary/Keyword: silk proteins

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Uptake of Fibroin Microspheres by 3T3 Cells (3T3 세포에 의한 피브로인 마이크로스피어의 흡수)

  • Lee, Jin Sil;Go, Nam Kyung;Lee, Shin Young;Hur, Won
    • KSBB Journal
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    • v.29 no.5
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    • pp.328-335
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    • 2014
  • Vehicle toxicity is one of the main obstacles for intracellular delivery of bioactive compounds. Silk fibroin is a natural polymer proven to have high biocompatibility since being used as suture material. In this report, fibroin microspheres were prepared without any chemical modification or cross-linking not to affect its biocompatibility. The microspheres were taken up by more than 90% of 3T3 cells. Cellular uptake continued after medium replenishment with a different-colored fluorescent microsphere, suggesting that simultaneous ingestion and exocytosis occurred. Cellular uptake of fibroin microspheres did not affect cell viability. Intracellular trafficking of the microspheres using lysosome-specific fluorescent dye revealed that fibroin microspheres were localized both in the cytoplasm and in the lysosome. Accordingly, fibroin microspheres can be a potential vehicle for intracytoplasmic delivery of large cargos, such as mixtures of proteins, nutrients or artificial organelles.

Antioxidant and Anti-Adipogenic Activities of Bread Containing Corn Silk, Job's Tears, Lentinus edodes, and Apple Peel in 3T3-L1 Preadipocytes (옥수수수염, 율무, 표고버섯 그리고 사과껍질을 함유한 빵의 항산화 및 3T3-L1 지방 전구세포 분화 억제 활성)

  • Lee, Chang Won;Park, Yong Il;Kim, Soo-Hyun;Lim, Heekyung;Chung, Mi Ja
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.45 no.5
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    • pp.651-663
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    • 2016
  • Corn silk, Job's tears, Lentinus edodes, and apple peel 70% ethanol extracts (CS, JT, LE, and AP) were studied for their antioxidant activities. CS among all extracts showed the highest antioxidant activities based on total polyphenol and flavonoid contents, 2,2-diphenyl-${\beta}$-picrylhydrazyl and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonate) radical scavenging activity, and reducing power. Adipocyte differentiation was investigated by Oil Red O staining assay using CS, JT, LE, AP, and extract of developed bread containing corn silk, Job's tears, Lentinus edodes, and apple peel (DB) treated to 3T3-L1 adipocytes. DB1 and DB2 showed anti-adipogenic and antioxidant effects. Triglyceride (TG) accumulation in 3T3-L1 cells was measured, and among the samples tested (CS, JT, LE, and AP), CS was found to have the highest inhibitory activity against TG accumulation of differentiated 3T3-L1 adipocytes and regulated factors associated with adipogenesis. CS suppressed lipid droplet formation and adipocyte differentiation in 3T3-L1 cells in a dose-dependent manner. We examined the effects of CS on the levels of CCAAT-enhancer-binding protein ${\beta}(C/EBP{\beta})$, peroxisome proliferator activated receptor ${\gamma}(PPAR{\gamma})$, and adipocyte-specific lipid binding protein (aP2) mRNA as well as protein levels in 3T3-L1 cells treated with CS at various concentrations (0, 10, 50, and $100{\mu}g/mL$) during adipocyte differentiation and treatment with CS in 3T3-L1 adipocytes down-regulated expression of $PPAR{\gamma}$ and aP2 mRNA. CS also significantly inhibited up-regulation of $C/EBP{\beta}$, $PPAR{\gamma}$, and aP2 proteins during adipocyte differentiation. These data indicate that DBs have anti-adipogenic activity induced by CS in 3T3-L1 preadipocytes, and CS exerts anti-adipogenic activity by inhibiting expression of $C/EBP{\beta}$, $PPAR{\gamma}$, and aP2 signaling pathway in 3T3-L1 adipocytes. JT, LE, and AP had no inhibitory effects on differentiation of 3T3-L1 preadipocytes but displayed strong antioxidant effects. These results suggest that the developed bread may be a health beneficial food that can prevent or treat obesity and diseases induced by oxidative stress.

Reeling of recombinant flourescence cocoons through low temperature decompressed cooking (저온감압 자견법에 의한 재조합 형광누에고치의 조사)

  • Park, Jong-Hwa;Kim, Sung-Wan;Jeong, Young-Hun;Lee, Jong-Kil;Go, Young-Mi;Lee, Sang-Chan;Choi, Kwang-Ho;Kim, Seong-Ryul;Goo, Tae-Won
    • Journal of Sericultural and Entomological Science
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    • v.51 no.2
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    • pp.142-146
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    • 2013
  • The fluorescent proteins are generally denatured by heat treatment and thus lose their color. The normal reeling method includes processing by drying and cooking the cocoons near $100^{\circ}C$ before reeling. Therefore, the usual processing method cannot be used for making colored fluorescent silks. To develop a method that is applicable to producing transgenic silk without color loss, we develop reeling methods adequate for a recombinant fluorescence cocoons. It was found that the fluorescence cocoons keep their native color when dried at temperatures lower than $60^{\circ}C$ for 15 h. Also, a new cooking method to soften the fluorescent cocoons was developed: the cocoons were soaked in a solution of 0.2% sodium carbonate ($Na_2CO_3$)/0.1% nonionic surfactant (Triton X100) at $60^{\circ}C$ and then placed under vacuum. The repeated vacuum treatments enabled complete penetration of the solution into the cocoons, and the cocoons were thus homogenously softened and ready for reeling. In this state, the cooked cocoons can be reeled by an automated reeling machine. Our results suggest that drying and cooking of the cocoons at low temperature enables the subsequent reeling of the colored fluorescent silks by an automatic reeling machine without color loss and can produce silks that can be used for making higher value-added silk materials.

Hepatic Steatosis Alleviated in Diabetic Mice upon Dietary Exposure to Fibroin via Transgenic Rice: Potential STAMP2 Involvement in Hepatocytes

  • Park, Ji-Eun;Jeong, Yeon Jae;Kim, Hye Young;Yoo, Young Hyun;Lee, Kwang Sik;Yang, Won Tae;Kim, Doh Hoon;Kim, Jong-Min
    • Development and Reproduction
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    • v.24 no.3
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    • pp.231-239
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    • 2020
  • Many benefits of silk protein fibroin (SPF) have been suggested in biomedical applications; and notably, significant SPF effects have been observed for metabolic syndromes that are directly linked to insulin resistance, such as type 2 diabetes mellitus (T2DM). Based on our previous findings, we believe that SPF from spiders exhibits outstanding glucose-lowering effects in diabetic BKS.Cg-m+/+Leprdb mice. In order to evaluate the dietary effects of SPF in diabetic animals, we generated several lines of transgenic rice (TR) that expresses SPF, and the feeding of TR-SPF to diabetic animals decreased blood glucose levels, but did not change insulin levels. Western blot analyses of hepatic proteins showed that AMP-activated protein kinase (AMPK) expression and phosphorylation both decreased in TR-SPF-fed groups, compared with controls. This finding suggests that the glucose-lowering effects in this diabetic animal model might be AMPK-independent. In contrast, six-transmembrane protein of prostate 2 (STAMP2) was upregulated after TR-SPF exposure. Together with STAMP2, the Akt protein phosphorylation increased after TR-SPF exposure, which indicates that STAMP2 leads to Akt phosphorylation and thus increases insulin sensitivity in hepatocytes. Importantly, the hepatic steatosis that was seen in the liver of diabetic mice was remarkably alleviated in TR-SPF-fed mice. Hepatocytes that were immunopositive for STAMP2 were overwhelmingly observed in hepatic tissues from TR-SPF-fed mice compared to the control. Taken together, these results suggest that feeding diabetic mice with TR-SPF upregulates STAMP2 expression and increases Akt phosphorylation in hepatic tissues and thus potentially alleviates insulin resistance and hepatic steatosis.

Analysis of nutrient composition of silkworm pupae in Baegokjam, Goldensilk, Juhwangjam, and YeonNokjam varieties

  • Lee, Ji Hae;Jo, You-Young;Kim, Seong-Wan;Kweon, HaeYong
    • International Journal of Industrial Entomology and Biomaterials
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    • v.43 no.2
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    • pp.85-93
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    • 2021
  • Silkworm pupae have been utilized as a food source and have high-quality proteins and fatty acids. However, studies on the nutritional composition of pupae according to their variety, developmental stage, and sex have not been conducted. In this study, the nutritional composition of four Korean silk varieties (Baegokjam, BG; Goldensilk, GS; Juhwangjam, JH; and YeonNokjam, YN) were analyzed according to developmental stage and sex. The main ingredient of the pupa was protein, and the protein levels were higher in females than in males. Fat levels were high in males and increased in YN and GS during the late stage but decreased in BG. Fiber content rapidly increased in the late stages, and the lowest content was observed in BG. The pupae contained all essential amino acids, which were detected at higher ratios in females with a high protein content. Fatty acids had a different predominance depending on the variety. Oleic acid level was high in BG, linoleic acid and linolenic acid levels were high in GS, and palmitic acid and stearic acid levels were high in JH. In the mineral analysis, zinc was dominant in BG, whereas calcium and iron levels were relatively low. Zinc level was low in GS, and calcium and magnesium levels were high in JH. Potassium, sodium, magnesium, and phosphorus levels were low in YN. The silkworm pupae showed differences in components according to their variety, sex, and developmental stage; therefore, selecting a variety suitable for its purpose is necessary.

Sericin Enhances Secretion of Thyroglobulin in the Thyrocytes (갑상선세포에서 sericin에 의한 thyroglobulin의 분비증가)

  • Jin, Cho-Yi;Song, Seong-Hee;Go, Young-Hwa;Kwon, Ki-Sang;Yun, Eun-Young;Goo, Tae-Won;Yeo, Joo-Hong;Kim, Seung-Whan;Choi, Jong-Soon;Yu, Kweon;Kwon, O-Yu
    • Journal of Life Science
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    • v.20 no.8
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    • pp.1249-1253
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    • 2010
  • Sericin is a type of high molecular weight water-soluble glycoprotein surrounding fibroin (silk protein) that has been used as a cell culture supplement and accelerates cell proliferation in various serum-free media. The purpose of this study was to investigate the enhancing effect of thyroglobulin (Tg) secretion by sericin in thyrocytes, FRTL-5 cells. While Tg-mRNA expression was not enhanced, a secreted form of Tg was obviously increased by sericin. In this status, expression of both endoplasmic reticulum (ER) molecular chaperones (Bip & calreticulin) and ER membrane proteins (IRE1, PERK & ATF6) was enhanced. The proximal step of IRE1, XBP1 mRNA splicing was slightly detected however, the proximal step of PERK, phosphorylation of $eIF2{\alpha}$, was changeless. In addition, sericin enhanced cell viability by the MTT assay. The above results showing the ability of sericin to promote protein production demonstrated its potential usefulness as a new biomaterial.

Non-Destructive Scientific Analysis of the Gold Fabric Excavated of Cheongsong Shim's Grave (청송심씨 묘에서 출토된 금직물의 비파괴 과학적 분석)

  • Lee, Hwang-Jo;Wi, Koang-Chul
    • Journal of Conservation Science
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    • v.38 no.3
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    • pp.243-253
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    • 2022
  • Using non-destructive analytical methods, we identified the material characteristics of two gold fabric artifacts excavated from the Cheongsong Sim clan (Bugeum Wonsam, Jikgeum Chima), including the artifact condition, fiber type, surface contamination, and metallic threads. We found that the artifacts were buried and had turned brown; thus, we were unable to determine their original color. The fiber type was determined to be silk from cocoons, based on scanning electron microscopy, Fourier transform infrared (FT-IR) analyses of Amide I, II, III, and IV peaks, and color reactions Further, the FT-IR and X-ray fluorescence (XRF) analyses identified the white and black stains as natural resin hydrolyzed substances, such as lipids and proteins, that occurred as microbial decomposition due to body decay. Finally, the XRF analyses identified the thin gold layer of the metallic yarn as gold (Au). According to the FT-IR data and the color reaction to the metallic yarn medium, the adhesive component of the medium was a product of-Amides I, II, III, and 3000 cm-1 within Amides A and B (an animal type), respectively. Thus, the medium was identified as Hanji (Korean paper), which is made from domestically produced Broussonetia kazinoki fibers.