• Title/Summary/Keyword: Zebrafish

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Ecotoxicity Assessment of Silver Nanomaterials with Different Physicochemical Characteristics in Diverse Aquatic Organisms (다양한 특성의 은나노물질이 수생생물에 미치는 독성영향평가)

  • Hong, Nam-Hui;Jung, Youn-Joo;Park, June-Woo
    • Korean Journal of Environmental Biology
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    • v.34 no.3
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    • pp.183-192
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    • 2016
  • Silver nanomaterials have been intensively applied in consumer products of diverse industrial sectors because of their strong biocidal properties and reported to be hazardous to aquatic organisms once released in the environment. Nanomaterials including sliver, are known to be different in toxicity according to their physicochemical characteristics such as size, shape, length etc. However studies comparing toxicity among silver nanomaterials with different physicochemical characteristics are very limited. Here, toxicities of silver nanomaterials with different size (50, 100, 150 nm), length (10, $20{\mu}m$), shape (wire, sphere), and coating material (polyvinylpyrrolidone, citrate) using OECD test guidelines were evaluated in aquatic species (zebrafish, daphnia, algae) and compared. On a size property, the smaller of silver nanomaterials, the more toxic to tested organisms. Sphered type of silver nanomaterials was less toxic to organisms than wired type, and shorter nanowires were less toxic than longer ones. Meanwhile the toxic effects of materials coated on silver nanomaterials were slightly different in each tested species, but not statistically significant. To the best of our knowledge, it is first investigation to evaluate and compare ecotoxicity of silver nanomaterials having different physicochemical characteristics using same test species and test guidelines. This study can provide valuable information for human and environmental risk assessment of silver nanomaterials and guide material manufacturers to synthesize silver nanomaterials more safely to human and environment.

Calmodulin of Olive Flounder Paralichthys olivaceus : Cloning and Expression Analysis

  • Hong, Gyeong-Eun;Kong, Hee Jeong;Nam, Bo-Hye;Kim, Young-Ok;Kim, Woo-Jin;Lee, Sang-Jun;Choi, Tae-Jin
    • Journal of Marine Bioscience and Biotechnology
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    • v.2 no.4
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    • pp.234-237
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    • 2007
  • Calmodulin (CaM) is a $Ca^{2+}$-binding protein essential for biological functions mediated through $Ca^{2+}$-dependent mechanism. A cDNA clone for CaM was isolated from a cDNA library of olive flounder Paralichthys olivaceus. The CaM cDNA concists of 782 bp and encodes a polypeptide of 149 amino acids with four $Ca^{2+}$-binding motifs EF-hands (EF-I, EF-II, EF-III, and EF-IV). The deduced amino acid sequence of CaM shows 97-100% amino acid sequence identity to other CaM sequences. Semi-quantitative PCR analysis revealed that the CaM transcription was began during early development and the CaM mRNA is expressed highly in brain and intestine, and moderately in kidney, gill, and eye of healthy olive flounder. Taken together, CaM may be necessary for early olive flounder development and that it may have a part in homeostasis.

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Cloning of the Bombyx mori short neuropeptide F receptor (BsNPF-R) cDNA (누에 short neuropeptide F receptor (BsNPF-R)의 cDNA cloning)

  • Shin, Hyojung;Kwon, Kisang;Hong, Sun Mee;Kim, Hong Geun;Park, Kwan-Ho;Choi, Ji-Young;Kim, Seung-Whan;Yu, Kweon;Kwon, O-Yu
    • Journal of Life Science
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    • v.26 no.6
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    • pp.721-726
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    • 2016
  • It has already been reported that short neuropeptide F (sNPF) stimulates feeding behaviors in a wide variety of insect species. In the present study, we cloned cDNA, encoding a sNPF receptor homologue from a silkworm, Bombyx mori, named BsNPF-R. The amino acid sequence of BsNPF-R was compared with those of sNPF-R thus far reported, which is shared with humans (36%), mice (34%), zebrafish (35%), and fruit flies (51%), respectively. A BsNPF-R protein’s mass was theoretically estimated to be 42,731 Da and it is a putative plasma membrane-penetrating protein. The mRNA expression of BsNPF-R was tested; the results showed that a strong expression was detected at the midgut, post-silk gland, Malpighian, and testis; however, a weak expression was at the fat body, hemocyte, and ovary. In addition, the synthesized sNPF of a silkworm regulated the BsNPF-R mRNA expression through the cell-based functional analysis.

Anthocyanins from Hibiscus syriacus L. Attenuate LPS-Induced Inflammation by Inhibiting the TLR4-Mediated NF-κB Signaling Pathway

  • Karunarathne, Wisurumuni Arachchilage Hasitha Maduranga;Molagoda, Ilandarage Menu Neelaka;Lee, Kyoung Tae;Choi, Yung Hyun;Kang, Chang-Hee;Jeong, Jin-Woo;Kim, Gi-Young
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2019.10a
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    • pp.92-92
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    • 2019
  • Excessive or chronic inflammation contributes to the pathogenesis of many inflammatory diseases such as sepsis, rheumatoid arthritis, and ulcerative colitis. Hibiscus syriacus L. has been used as a medicinal plant in many Asian countries, even though its anti-inflammatory activity has been unclear. Therefore, we investigated the anti-inflammatory effect of anthocyanin fractions from the H. syriacus L. varieties Pulsae (PS) on the lipopolysaccharide (LPS)-induced expression of proinflammatory mediators and cytokines in RAW264.7 macrophages. PS suppressed LPS-induced nitric oxide (NO) and prostaglandin $E_2$ ($PGE_2$) secretion concomitant with downregulation of inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2) expression. Furthermore, PS inhibited the production of proinflammatory cytokines such as tumor necrosis factor-alpha ($TNF-{\alpha}$), interleukin-6 (IL-6), and IL-12 in LPS-stimulated RAW264.7 macrophages. Further study showed that PS significantly decreased LPS-induced nuclear translocation of the nuclear $factor-{\kappa}B$ ($NF-{\kappa}B$) subunits, p65 and p50. Molecular docking data showed that many anthocyanins from PS fit into the hydrophobic pocket of MD2 and bound to Toll-like receptor 4 (TLR4), indicating that PS inhibits the TLR4-MD2-mediated inflammatory signaling pathway. Especially, apigenin-7-O-glucoside most powerfully bound to MD2 and TLR4 through LYS122, LYS122, and SER127 at a distance of $2.205{\AA}$, $3.098{\AA}$, and $2.844{\AA}$ and SER441 at a distance of $2.873{\AA}$ (docking score: -8.4) through hydrogen bonding, respectively. Additionally, PS inhibited LPS-induced TLR4 dimerization/expression on the cell surface, which consequently decreased MyD88 recruitment and IRAK4 phosphorylation. PS completely blocked LPS-mediated mortality in zebrafish larvae by diminishing the recruitment of neutrophil and macrophages accompanied by low levels of proinflammatory cytokines. Taken together, our results indicate that PS attenuates LPS-mediated inflammation in both in vitro and in vivo by blocking the TLR4/MD2-MyD88/IRAK4-$NF-{\kappa}B$ axis. Therefore, PS might be used as a novel modulatory candidate for effective treatment of LPS-mediated inflammatory diseases.

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Cloning and Characterization of Phosphoinositide 3-Kinase γ cDNA from Flounder (Paralichthys olivaceus) (넙치에서 분리된 phosphoinositide 3-kinase γ 유전자의 클로닝 및 특성 연구)

  • Jeong, Tae Hyug;Youn, Joo Yeon;Ji, Keunho;Seo, Yong Bae;Kim, Young Tae
    • Journal of Life Science
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    • v.24 no.4
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    • pp.343-351
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    • 2014
  • Phosphoinositide 3-kinase (PI3K) plays a central role in cell signaling and leads to cell proliferation, survival, motility, exocytosis, and cytoskeletal rearrangements, as well as specialized cell responses, superoxide production, and cardiac myocyte growth. PI3K is divided into three classes; type I PI3K is preferentially expressed in leukocytes and activated by ${\beta}{\gamma}$ subunits of heterotrimeric G-proteins. In this study, the cDNAs encoding the $PI3K{\gamma}$ gene were isolated from a brain cDNA library constructed using the flounder (Paralichthys olivaceus). The sequence of the isolated $PI3K{\gamma}$ was 1341 bp, encoding 447 amino acids. The nucleotide sequence of the $PI3K{\gamma}$ gene was analyzed with that of other species, including Oreochromis niloticus and Danio rerio, and it turned out to be well conserved during evolution. The $PI3K{\gamma}$ gene was subcloned into the expression vector pET-44a(+), and expressed in the E. coli BL21 (DE3) codon plus cell. The resulting protein was expressed as a fusion protein of approximately 49 kDa containing a C-terminal six-histidine extension that was derived from the expression vector. The expressed protein was purified to homogeneity by His-tag affinity chromatography and showed enzymatic activity corresponding to $PI3K{\gamma}$. The binding of wortmannin to $PI3K{\gamma}$, as detected by anti-wortmannin antisera, closely followed the inhibition of the kinase activities. The results obtained from this study will provide a wider base of knowledge on the primary structure and characterization of the $PI3K{\gamma}$ at the molecular level.

Studies on Antioxidant Activity and In Vitro Inhibitory Activity of Tyrosinase and Collagenase in Artocarpus nitidus subsp. lingnaensis (Merr.) F.M. Jarrett using 4 Parameter Logistic (변수 분석을 통한 아토카푸스 니티두스 추출물과 분획물의 항산화, 타이로시나제 및 콜라제나제 In Vitro 저해활성 연구)

  • Son, Kwang-Hee;Kim, Young Kook;Choi, Sangho;Zhang, Zhiyun;Shin, Dong-Ha;Lee, Jong Suk;Park, Ho-Yong
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.45 no.2
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    • pp.161-173
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    • 2019
  • In this study, the antioxidative and inhibitory activity of tyrosinase and collagenase for the solvent extract and silica column fractions of Artocarpus nitidus were evaluated. The activities were quantified using the 4 parameter logistic. LC/MS analysis showed that the major component of the fractions was polyphenol and the total polyphenol content of the extract was $48.1{\pm}2.6mg\;GAE/g$. The radical scavenging activities ($SC_{50}$) for 1,1-diphenyl-2-picrylhydrazyl of the extract, fraction-1 and fraction-2 were 16.7, 42.0 and $10.1{\mu}g/mL$, respectively. The value for fraction-2 was the closest to ascorbic acid ($1.5{\mu}g/mL$). The tyrosinase inhibitory activity of the extracts and the fractions showed $IC_{50}$ of 64.9, 0.9 and $1.2{\mu}g/mL$, respectively, and overall activity was higher than that of kojic acid ($7.4{\mu}g/mL$) and arbutin ($119.0{\mu}g/mL$). In the experiment by zebrafish embryo, the whitening activity of fraction-2 (27.5%) was higher than that of kojic acid (18.6%), and there was no adverse effect up to $500{\mu}g/mL$ of fraction-2. For the collagenase inhibitory activity, the samples showed $IC_{50}$ of 139.8, 20.6, and $16.8{\mu}g/mL$, respectively, which were competitive to 1, 10-Phenanthroline ($55.4{\mu}g/mL$). The extract and fraction-2 showed $IC_{50}$ of 61.8 and $67.1{\mu}g/mL$ for elastase. These results suggest that A. nitidus extract can be used as a cosmetic material useful for antioxidant, whitening, and prevention of skin aging without adverse effects.

Molecular Characterization and Expression Analysis of Clathrin-Associated Adaptor Protein 3-δ Subunit 2 (AP3S2) in Chicken

  • Oh, Jae-Don;Bigirwa, Godfrey;Lee, Seokhyun;Song, Ki-Duk
    • Korean Journal of Poultry Science
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    • v.46 no.1
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    • pp.31-37
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    • 2019
  • A chicken clathrin-associated adaptor protein $3-{\delta}$ subunit 2 (AP3S2) is a subunit of AP3, which is involved in cargo protein trafficking to target membrane with clathrin-coated vesicles. AP3S2 may play a role in virus entry into host cells through clathrin-dependent endocytosis. AP3S2 is also known to participate in metabolic disease developments of progressions, such as liver fibrosis with hepatitis C virus infection and type 2 diabetes mellitus. Chicken AP3S2 (chAP3S2) gene was originally identified as one of the differentially expressed genes (DEGs) in chicken kidney which was fed with different calcium doses. This study aims to characterize the molecular characteristics, gene expression patterns, and transcriptional regulation of chAP3S2 in response to the stimulation of Toll-like receptor 3 (TLR3) to understand the involvement of chAP3S2 in metabolic disease in chicken. As a result, the structure prediction of chAP3S2 gene revealed that the gene is highly conserved among AP3S2 orthologs from other species. Evolutionarily, it was suggested that chAP3S2 is relatively closely related to zebrafish, and fairly far from mammal AP3S2. The transcriptional profile revealed that chAP3S2 gene was highly expressed in chicken lung and spleen tissues, and under the stimulation of poly (I:C), the chAP3S2 expression was down-regulated in DF-1 cells (P<0.05). However, the presence of the transcriptional inhibitors, BAY 11-7085 (Bay) as an inhibitor for nuclear factor ${\kappa}B$ ($NF{\kappa}B$) or Tanshinone IIA (Tan-II) as an inhibitor for activated protein 1 (AP-1), did not affect the expressional level of chAP3S2, suggesting that these transcription factors might be dispensable for TLR3 mediated repression. These results suggest that chAP3S2 gene may play a significant role against viral infection and be involved in TLR3 signaling pathway. Further study about the transcriptional regulation of chAP3S2 in TLR3 pathways and the mechanism of chAP3S2 upon virus entry shall be needed.