• Title/Summary/Keyword: Cell nucleus

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Molecular probe for identification of cysts of resting cyst of PSP-producer Alexandrium tamarense (Dinophyceae) (분자생물학적 방법을 이용하여 마비성 패류 독소를 생산하는 알렉산드륨 타마렌스 시스트 탐색)

  • Cho, Eun-Seob
    • Journal of Life Science
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    • v.13 no.2
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    • pp.163-167
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    • 2003
  • Identification of species within the toxin-producing genus Alexandrium is vital for biotoxin monitoring and mitigation decisions regarding shellfish industry. In particular, the discrimination of resting cysts of only A. tamarense from that of Alexandrium spp. is considerable important to fundamentally monitor and predict this species before vegetative cells occur in the nature. Fluorescent cTAM-F1 DNA probe was responsible to not only binding the activity of the vegetative cells in A. tamarense, but also to the resting cysts, which was treated with methanol after fixation and stained by primuline on the surface The location of fluorescence in cultured vegetative cells and resting cysts was almost at tile bottom of the nucleus. The optimal incubation temperature and time using in situ hybridization were 50-$54^{\circ}C$ and 40-60 min, respectively, to penetrate the DNA probe into cell.

Ultrastructural Changes of Oocytes from Vitellogenesis into Maturation in Korean Catfish, Silurus asotus (난황형성부터 성숙까지 메기(Silurus asotus) 난모세포의 미세구조)

  • Yoon, Jong-Man;Kim, Young-Gill
    • Applied Microscopy
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    • v.28 no.4
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    • pp.447-463
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    • 1998
  • This study was carried out to investigate the histomorphological changes and the electrophoretic patterns of egg components, obtained from 100 of 2-year-old female catfishes (Silurus asotus). Female Korean catfishes collected in the vicinity of Chollabukdo have synchronous ovaries which discontinously ovulate eggs during the breeding season (from late May until early July). The fishes were killed by severing the spinal cord just posterior to the head after immobilization with tricaine methanesulfonate (MS 222). Especially, the light microscopic and ultrastructural changes of ooplasm and follicular membranes of oocytes, were examined by means of light and transmission electron microscopy. The size of the nucleoli and number of the yolk granules increased as the oocyte groved. Yolk granules were deposited in the oocyte as fluid Due to tile presence of large early and late maturing oocytes, their ovaries were enlarged, transparent, granular and greenish in color. As the percentages of fish in late maturing oocyte (LMO) and ripe oocyte (RO) stage increased from March to April, mean gonadosomatic index (GSI) values (19.95%) increased. Zona radiata changed from squamous into cuboid in shape in the early maturing oocyte (EMO) stage. Processes, microvilli, from the zona radiata and from the oocyte grow, and make contact with each other in the pore canals of the zona radiata during vitellogenesis, but are withdrawn as the zona radials becomes more compact and devoid of pore canals during oocyte maturation. Seasonal changes in the microscopic appearance of the ovaries were well correlated with those in both gonadosomatic index and macroscopic appearance. The main cytological changes such as increase in size of cell, nucleus, nucleolus, and increase in number of nucleoli and mitochondria demonstrated with electron microscopy in the previtellogenic oocytes of Korean catfish, provided evidences for important synthetic processes in an early preparatory phase of oocyte development. The electrophoretic pattern of major band in mature stage was much thicker (24 k, 66 k, 90-110k dalton) than that in previtellogenic phase.

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Effect of Hwangryunagyotang Water Extract on Endothelial Cells by Free Cholesterol. (황련아교탕(黃連阿膠湯)이 Free Cholesterol에 의한 혈관내피세포 손상에 미치는 영향)

  • Lee, So-Yeon;Yoon, Hyun-Duk;Shin, Oh-Chul;Shin, Yoo-Jung;Park, Chi-Sang
    • The Journal of Internal Korean Medicine
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    • v.27 no.3
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    • pp.589-602
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    • 2006
  • Hwangryunagyotang is supposed to have significant effects on some sorts of cardiovascular diseases like atherosclerosis. For this study. ACAT inhibitor was put in LDLR -/- mice to derive free cholesterol from it. This was to examine the effectiveness of Hwangryunnagyotang on its protecting and recovering function with endothelial cells damaged by free cholesterol through experimental. The results reported below. Hwangryunagyotang suppressed the crystallization of reactive oxygen species in macrophages and the numbers of free cholesterol crystal plate structured and reduced fragmentation of nucleus in ECV 304 cell strain by ACAT inhibitor significantly. Hwangryunagyotang also suppressed the necrosis of tissue in LDLR -/- mice' (treated with ACAT inhibitor) inflammatory portion which is adjacent to aortic root, proximal aorta and carotid artery by immunohistochemistry and fluorescence microscopy. On the whole, Hwangryunagyotang suppressed the necrosis of endothelial cells and especially it's effcet for the necrosis of para-myocardial tissues by free cholesterol. With this result, I suggest Hwangryunagyotang might have protective and recovery effects on atherosclerosis, so we need to carry on this study henceforth clinically and experimentally as well.

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Inhibition of Nitric Oxide Production by Ethyl Digallates Isolated from Galla Rhois in RAW 264.7 Macrophages

  • Park, Pil-Hoon;Hur, Jin;Lee, Dong-Sung;Kim, Youn-Chul;Jeong, Gil-Saeng;Sohn, Dong-Hwan
    • Biomolecules & Therapeutics
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    • v.19 no.4
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    • pp.419-424
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    • 2011
  • Galla Rhois and its components are known to possess anti-infl ammatory properties. In the present study, we prepared equilibrium mixture of ethyl m-digallate and ethyl p-digallate isomers (EDG) from Galla Rhois and examined its effect on nitric oxide (NO) production in murine macrophage cell line. Treatment of RAW264.7 macrophages with EDG signifi cantly inhibited NO production and inducible nitric oxide synthase (iNOS) expression stimulated by LPS, as assessed by Western blot and quantitative RT-PCR analyses. We also demonstrated that EDG treatment led to an increase in heme oxygenase-1 (HO-1) mRNA and protein expression. EDG treatment also enhanced expression level of nuclear factor-erythroid 2-related factor 2 (Nrf2) in nucleus, which is critical for transcriptional induction of HO-1. Treatment with SnPP (tin protoporphyrin IX), a selective HO-1 inhibitor, reversed EDG-mediated inhibition of nitrite production, suggesting that HO-1 plays an important role in the suppression of NO production by EDG. Taken together, these results indicate that EDG isolated from Galla Rhois suppresses LPS-stimulated NO production in RAW 264.7 macrophages via HO-1 induction.

OCT4B Isoform Promotes Anchorage-Independent Growth of Glioblastoma Cells

  • Choi, Sang-Hun;Kim, Jun-Kyum;Jeon, Hee-Young;Eun, Kiyoung;Kim, Hyunggee
    • Molecules and Cells
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    • v.42 no.2
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    • pp.135-142
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    • 2019
  • OCT4, also known as POU5F1 (POU domain class 5 transcription factor 1), is a transcription factor that acts as a master regulator of pluripotency in embryonic stem cells and is one of the reprogramming factors required for generating induced pluripotent stem cells. The human OCT4 encodes three isoforms, OCT4A, OCT4B, and OCT4B1, which are generated by alternative splicing. Currently, the functions and expression patterns of OCT4B remain largely unknown in malignancies, especially in human glioblastomas. Here, we demonstrated the function of OCT4B in human glioblastomas. Among the isoform of OCT4B, OCT4B-190 ($OCT4B^{19kDa}$) was highly expressed in human glioblastoma stem cells and glioblastoma cells and was mainly detected in the cytoplasm rather than the nucleus. Overexpression of $OCT4B^{19kDa}$ promoted colony formation of glioblastoma cells when grown in soft agar culture conditions. Clinical data analysis revealed that patients with gliomas that expressed OCT4B at high levels had a poorer prognosis than patients with gliomas that expressed OCT4B at low levels. Thus, $OCT4B^{19kDa}$ may play a crucial role in regulating cancer cell survival and adaption in a rigid environment.

Study on the Antagonistic Activity on Arylhydrocarbon Receptor of Phenyldiazenylphenylpicolinamides (Phenyldiazenylphenylpicolinamide 유도체들의 방향성탄화수소 수용체의 길항 활성에 대한 연구)

  • Yoon, Wan-Young;Lee, Hyosung
    • Journal of Digital Convergence
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    • v.17 no.1
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    • pp.443-447
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    • 2019
  • Aryl hydrocarbon receptor (AhR) is the master regulator of xenobiotics metabolizing enzymes (XMEs). AhR is activated by aryl hydrocarbons upon binding then goes into the cell nucleus and acts as a transcription factor. Despite the role of AhR in human physiology has been investigated for a long while, it is yet to be understood mainly due to the lack of appropriate chemical agents. Furthermore, it has been reported that AhR is related to a wide range of pathogenesis. In addition, recent studies suggest that the study on the development of AhR antagonist may provide a valid therapeutic agent. Some known antagonists in current use are partially agonistic whereas a pure antagonist is still absent. In this study, two phenyl-ring structures of phenyldiazenylphenylpicolinamide has been modified into various structures and evaluated its impact on the AhR antagonistic activity to elucidate the structure-activity relationship.

L-ascorbic acid induces apoptosis in human laryngeal epidermoid Hep-2 cells by modulating the nuclear factor kappa-light-chain-enhancer of activated B cells/mitogen-activated protein kinase/Akt signaling pathway

  • Park, Jung-Sun;Kim, Yoon-Jung;Park, Sam Young;Chung, Kyung-Yi;Oh, Sang-Jin;Kim, Won-Jae;Jung, Ji-Yeon
    • International Journal of Oral Biology
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    • v.45 no.4
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    • pp.169-178
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    • 2020
  • L-ascorbic acid (L-AA; vitamin C) induces apoptosis in cancer cells. This study aimed to elucidate the molecular mechanisms of L-AA-induced apoptosis in human laryngeal epidermoid carcinoma Hep-2 cells. L-AA suppressed the viability of Hep-2 cells and induced apoptosis, as shown by the cleavage and condensation of nuclear chromatin and increased number of Annexin V-positive cells. L-AA decreased Bcl-2 protein expression but upregulated Bax protein levels. In addition, cytochrome c release from the mitochondria into the cytosol and activation of caspase-9, -8, and -3 were enhanced by L-AA treatment. Furthermore, apoptosis-inducing factor (AIF) and endonuclease G (EndoG) were translocated into the nucleus during apoptosis of L-AA-treated Hep-2 cells. L-AA effectively inhibited the constitutive nuclear factor-κB (NF-κB) activation and attenuated the nuclear expression of the p65 subunit of NF-κB. Interestingly, L-AA treatment of Hep-2 cells markedly activated Akt and mitogen-activated protein kinase (MAPK; extracellular signal-regulated kinase 1/2, p38, and c-Jun N-terminal kinase [JNK]) and and LY294002 (Akt inhibitor), SB203580 (p38 inhibitor) or SP600125 (a JNK inhibitor) decreased the levels of Annexin V-positive cells. These results suggested that L-AA induces the apoptosis of Hep-2 cells via the nuclear translocation of AIF and EndoG by modulating the Bcl-2 family and MAPK/Akt signaling pathways.

Anti-inflammatory and antioxidant effects of Barringtonia augusta Kurz extract (Barringtonia augusta Kurz 추출물의 항염증 및 항산화 효능 평가)

  • Ryu, Soo Ho;Kim, Min Jeong;Bach, Tran The;Jung, Sung Keun
    • Korean Journal of Food Science and Technology
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    • v.53 no.2
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    • pp.154-159
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    • 2021
  • Barringtonia augusta Kurz is a species of the genus Barringtonia. Although several studies have analyzed the biological activity of B. racemosa Roxb and B. acutangula, the anti-inflammatory and antioxidant effects of B. augusta extract (BKE) remain unclear. Therefore, in this study, we investigated the anti-inflammatory and antioxidant effects of BKE using lipopolysaccharide (LPS) and RAW 264.7. BKE suppressed LPS-induced nitric oxide (NO) and inducible NO synthase expression without affecting RAW 264.7 cell viability. Additionally, BKE showed 2,2-Diphenyl-1-picrylhydrazyl and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) radical scavenging capacities and inhibited LPS-induced reactive oxygen species production in RAW 264.7 cells. BKE also suppressed LPS-induced phosphorylation of IκB kinase and nuclear factor kappa-B (NF-κB) and p65 translocation from the cytosol to the nucleus in RAW 264.7 cells. These results suggest that BKE is a possible novel material that exerts beneficial antioxidant and anti-inflammatory effects through the inhibition of NF-κB signaling pathways.

M6A reader hnRNPA2/B1 is essential for porcine embryo development via gene expression regulation

  • Kwon, Jeongwoo;Jo, Yu-Jin;Yoon, Seung-Bin;You, Hyeong-ju;Youn, Changsic;Kim, Yejin;Lee, Jiin;Kim, Nam-Hyung;Kim, Ji-Su
    • Journal of Animal Reproduction and Biotechnology
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    • v.37 no.2
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    • pp.121-129
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    • 2022
  • Heterogeneous nuclear ribonucleoprotein A2/B1 (hnRNPA2/B1) is an N6-methyladenosine (m6A) RNA modification regulator and a key determinant of prem-RNA processing, mRNA metabolism and transportation in cells. Currently, m6A reader proteins such as hnRNPA2/B1 and YTHDF2 has functional roles in mice embryo. However, the role of hnRNPA2/B1 in porcine embryogenic development are unclear. Here, we investigated the developmental competence and mRNA expression levels in porcine parthenogenetic embryos after hnRNPA2/B1 knock-down. HhnRNPA2/B1 was localized in the nucleus during subsequent embryonic development since zygote stage. After hnRNPA2/B1 knock-down using double stranded RNA injection, blastocyst formation rate decreased than that in the control group. Moreover, hnRNPA2/B1 knock-down embryos show developmental delay after compaction. In blastocyste stage, total cell number was decreased. Interestingly, gene expression patterns revealed that transcription of Pou5f1, Sox2, TRFP2C, Cdx2 and PARD6B decreased without changing the junction protein, ZO1, OCLN, and CDH1. Thus, hnRNPA2/B1 is necessary for porcine early embryo development by regulating gene expression through epigenetic RNA modification.

Nrf2-mediated activation of HO-1 is required in the blocking effect of compound K, a ginseng saponin metabolite, against oxidative stress damage in ARPE-19 human retinal pigment epithelial cells

  • Cheol Park;Hee-Jae Cha;Kyoung-Seob Song;Heui-Soo Kim;EunJin Bang;Hyesook Lee;Cheng-Yun Jin;Gi-Young Kim;Yung Hyun Choi
    • Journal of Ginseng Research
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    • v.47 no.2
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    • pp.311-318
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    • 2023
  • Background: The beneficial effects of compound K (CK) on different chronic diseases have been shown to be at least related to antioxidant action. Nevertheless, since its antioxidant activity in human retinal pigment epithelial (RPE) cells is still unknown, here we investigated whether CK alleviates oxidative stress-stimulated damage in RPE ARPE-19 cells. Methods: The cytoprotective consequence of CK in hydrogen peroxide (H2O2)-treated cells was evaluated by cell viability, DNA damage, and apoptosis assays. Fluorescence analysis and immunoblotting were performed to investigate the inhibitory action of CK on reactive oxygen species (ROS) production and mitochondrial dysfunction. Results: H2O2-promoted cytotoxicity, oxidative stress, DNA damage, mitochondrial impairment, and apoptosis were significantly attenuated by CK in ARPE-19 cells. Furthermore, nuclear factor erythroid 2-related factor 2 (Nrf2) phosphorylation level and its shuttling to the nucleus were increased, which was correlated with upregulated activation of heme oxygenase-1 (HO-1). However, zinc protoporphyrin, a blocker of HO-1, significantly abrogated the preventive action of CK in H2O2-treated ARPE-19 cells. Conclusion: This study indicates that activation of Nrf2/HO-1 signaling by CK plays an important role in rescuing ARPE-19 cells from oxidative cellular damage.