• 제목/요약/키워드: intact cells

검색결과 444건 처리시간 0.028초

Antitumor activities of hypericin as a protein tyrosine kinase blocker

  • Kil, Kwang-Sup;Yum, Young-Na;Seo, Seung-Hoon;Lee, Kyung-Tae
    • Archives of Pharmacal Research
    • /
    • 제19권6호
    • /
    • pp.490-496
    • /
    • 1996
  • Naphtodianthrone hypericin produced a potent antitumor activity in vitro against several tumor cells. However, it did not show any cytotoxicity on normal cells such as Macaccus rheus monkey kidney cells (MA-104) and primary cultured rat hepatocytes up to $500{\mu}M$ concentration. Hypericin added to A431 human epidermoid carcinoma cell membrane inhibited the autophosphorylation of the epidermal growth factor (EGF) receptor and the tyrosine phosphorylation of RR-SRC peptide catalyzed by an EGF-receptor. Similarly, treatment of the A431 cells with hypericin inhibited the tyrosine phosphorylation of EGF-dependent endogenous EGF-receptor by western blotting analysis. Hypericin also inhibited the T cell PTK, $P56^{lck}$, in a dose-dependent fashion with an $IC_{50}=5{\mu}M$. The tyrosine phosphorylation, on RR-SRC peptide and EGF-induced receptor autophosphorylation, either in vitro or in intact cells was inhibited by hypericin at the same concentration as that in A431 cell proliferation. These data suggest that hypericin directly inhibits EGF-receptor and $P56^{lck}$ PTK activity in vitro and can mediate such action in vivo.

  • PDF

도축한우에 있어서 근육지방증 (Steatosis in a Slaughtered Korean Native Cattle)

  • Do, Sun-Hee;Lee, Cha-Soo;Jeong, Won-Il;Chung, Jae-Yong;Jeong, Da-Hee;Noh, Dong-Hyung;An, Mi-Young;Jee, Young-Heun;Lee, Mi-Na
    • 한국임상수의학회지
    • /
    • 제19권3호
    • /
    • pp.350-352
    • /
    • 2002
  • 2002년 5월 도축중 발견된 거세수소의 근육 병변에 대한 검사 의뢰가 들어왔다. 이 병변의 육안적 소견은 광범위한 근육의 지방침윤이었고, 병리조직학적 검사에서 근섬유가 지방조직에 의해 대치된 것이 관찰되었으며, 기타 염증 소견은 관찰되지 않았다. 본 증례는 골격근의 광범위한 부분이 지방조직으로 대치된 지방증(steatosis)으로 진단되어 이에 보고하는 바이다.

Immuno-modulation Effects of Ginsenoside Rg1 in Rat microglia

  • Joo, Seong-Soo;Won, Tae-Joon;Lee, Do-Ik
    • 대한약학회:학술대회논문집
    • /
    • 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.1
    • /
    • pp.204.2-205
    • /
    • 2003
  • Microglial cell is a monocyte involved in the brain, which acts for a primary immune reaction and phagocytosis. Microglia has also been considered to have a great role in AD pathogenesis due to its intact inflammatory and phagocytic responses against foreign invaders. In the study, we tried to investigate the modulation of activation of microglia using Rg1, a class of ginsenoside from red ginseng. which are known to protect neuron cells. (omitted)

  • PDF

Fucoidan Induces Apoptosis in A2058 Cells through ROS-exposed Activation of MAPKs Signaling Pathway

  • Ryu, Yea Seong;Hyun, Jin Won;Chung, Ha Sook
    • Natural Product Sciences
    • /
    • 제26권3호
    • /
    • pp.191-199
    • /
    • 2020
  • Fucoidan, a natural component of brown seaweed, has various biological activities such as anti-cancer activity, anti-oxidant, and anti-inflammatory against various cancer cells. However, the fucoidan has been implicated in melanoma cells via apoptosis signaling pathway. Therefore, we investigated apoptosis with fucoidan in A2058 human melanoma cells with dose- and time-dependent manners. In our results, A2058 cells viability decreased at relatively short-time and low-concentration through fucoidan. This effects of fucoidan on A2058 cells appeared to be mediated by the induction of apoptosis, as manifested by morphological changes through DNA-binding dye Hoechst 33342 staining. When a dose of 80 ㎍/mL fucoidan was treated, the cells were observed: crescent or ring-like structure, chromatin condensation, and nuclear fragmentation. With the increase at 100 ㎍/mL fucoidan, the cell membrane is intact throughout the total process, including membrane blebbing and loss of membrane integrity as well as increase of sub-G1 DNA. Furthermore, to understand the exact mechanism of fucoidan-treated in A2058 cells, western blotting was performed to detect apoptosis-related protein expression. In this study, Bcl-2 family proteins can be regulated by fucoidan, suggesting that fucoidan-induced apoptosis is modulated by intrinsic pathway. Therefore, expression of Bcl-2 and Bax may result in altered permeability, activating caspase-3 and caspase-9. And the cleaved form of poly ADP-ribose polymerase was detected in fucoidan-treated A2058 cells. These results suggest that A2058 cells are highly sensitive to growth inhibition by fucoidan via apoptosis, as evidenced by activation of extracellular signal-regulated kinases/p38/Bcl-2 family signaling, as well as alteration in caspase-9 and caspase-3.

Effect of growth phase of cyanobacterium on release of intracellular geosmin from cells during microfiltration process

  • Matsushita, Taku;Nakamura, Keisuke;Matsui, Yoshihiko;Shirasaki, Nobutaka
    • Membrane and Water Treatment
    • /
    • 제6권3호
    • /
    • pp.225-235
    • /
    • 2015
  • During low-pressure membrane treatments of cyanobacterial cells, including microfiltration (MF) and ultrafiltration (UF), there have reportedly been releases of intracellular compounds including cyanotoxins and compounds with an earthy-musty odor into the water, probably owing to cyanobacterial cell breakage retained on the membrane. However, to our knowledge, no information was reported regarding the effect of growth phase of cyanobacterial cells on the release of the intracellular compounds. In the present study, we used a geosmin-producing cyanobacterium, Anabaena smithii, to investigate the effect of the growth phase of the cyanobacterium on the release of intracellular geosmin during laboratory-scale MF experiments with the cells in either the logarithmic growth or stationary phase. Separate detection of damaged and intact cells revealed that the extent of cell breakage on the MF membrane was almost the same for logarithmic growth and stationary phase cells. However, whereas the geosmin concentration in the MF permeate increased after 3 h of filtration with cells in the logarithmic growth phase, it did not increase during filtration with cells in the stationary phase: the trend in the geosmin concentration in the MF permeate with time was much different between the logarithmic growth and stationary phases. Adsorption of geosmin to algogenic organic matter (AOM) retained on the MF membrane and/or pore blocking with the AOM were greater when the cells were in the stationary phase versus the logarithmic growth phase, the result being a decrease in the apparent release of intracellular geosmin from the stationary phase cells. In actual drinking water treatment plants employing membrane processes, more attention should be paid to the cyanobacterial cells in logarithmic growth phase than in stationary phase from a viewpoint of preventing the leakage of intracellular earthy-musty odor compounds to finished water.

기니피그 위점막이 카테콜아민 유발 위운동에 미치는 영향 (Influences of Gastric Mucosa upon the Catecholamine Induced Gastric Motility in Guinea-pig)

  • 이풍렬;김기환;이상진
    • The Korean Journal of Physiology
    • /
    • 제23권2호
    • /
    • pp.277-289
    • /
    • 1989
  • The effects of noradrenaline on the contractile and electrical activities were investigated using the circular muscle strips with intact mucosa prepared from the antrum and fundus of guinea-pig stomach. Electrical responses of circular muscle cells were recorded using glass capillary microelectrodes filled with 3 M KCI. All experiments were performed in tris-buffered Tyrode solution which was aerated with 100% $O_2\;and\;kept\;at\;35^{\circ}C$. The results obtained were as follows: 1) The spontaneous contractions recorded from the antral and fundic circular muscle strips with intact mucosa were suppressed dose-dependently by the application of noradrenaline, whereas those recorded from the mucosa-free strips were potentiated in a dose-dependent manner. 2) The inhibitory influences on the contractile activities in the normal intact strips were developed via both ${\alpha}-adrenoceptors\;and\;{\beta}-adrenoceptors$, while the excitatory influences in the mucosa-free strips resulted from the strong excitatory effect via ${\alpha}-adrenoceptors$ and the weak inhibitory effect via ${\beta}-adrenoceptors$. 3) Noradrenaline produced hyperpolarization of membrane potential, and increased the amplitude and the maximum rate of rise of slow waves in the mucosa-free strips of antral and fundic circular muscle. 4) Apamin blocked the appearance of the component of initial suppression of spontaneous phasic contractions observed in the mucosa-free strips of antral circular muscle after the application of noradrenaline. 5) The inhibitory influences on the contractile activities in the normal strips with intact mucosa remained unaffected even in the strip with separate mucosa, in which mucosa and muscle layer were mechanically disconnected . From the above results, following conclusions could be made. (1) There are no regional differences between the effects of noradrenaline on the antral circular muscle and those on the fundic circular muscle. (2) Excitatory responses to noradrenaline observed in the mucosa-free strip result from the dominant ${\alpha}-excitatory$ and tile weak ${\beta}-inhibitory$ action of noradrenaline. (3) Inhibitory responses to noradrenaline in the normal strips with intact mucosa develop via both ${\alpha}-inhibitory\;and\;{\beta}-inhibitory$ actions.

  • PDF

흰쥐 뇌내(腦內)의 무수카린성 콜린 수용체의 이질성(異質性) (Multiple Binding Affinities for Muscarinic Acetylcholine Receptors in Rat Brain)

  • 이종화
    • 대한약리학회지
    • /
    • 제23권2호
    • /
    • pp.101-111
    • /
    • 1987
  • 중추신경계 특히 뇌내(腦內)의 무수카린성 콜린 수용체 (mAchR)에 대한 수용체 특성의 연구의 하나로, 물리 화학적 성상에 다른 두 종류의 콜린길항제를 사용하여 서로 다른 두 형태의 조직에서 약물의 작용양상 및 다른 약물과의 상호작용을 정초하였다. 실험동물로는 흰쥐를 일정기간 규정사료를 사육하였고, 사용한 Radioactive ligands는 $(^3H)$ QNB와 $(^3H)$ NMS였으며 그외에 다른 수종의 길항제 또는 효능제와의 치환작용을 brain homogenates와 intact brain cell aggregates에서 관찰하여 다음과 같은 결과를 얻었다. 1. $(^3H)$ QNB와 $(^3H)$ NMS는 모두 질량작용의 법칙에 비례하여 수용체와의 결합에서 높은 친화력과 포화를 보였으며 또한 높은 결합 능력을 나타내었다. 더욱이 homogenates 제제와 intact cell aggregates제제에서의 결과 사이에는 유사한 점이 많았다. 2. Homogenates제제를 사용한 실험에서, 제 3 급아민콜린길항제인 QNB, atropine과 scopolamine 또는 제 4 급 암모늄골린 길항제인 methylatropine과 methylscopolamine을 사용하여 위의 radioactive ligands와의 치환작용을 검토하였다. $(^3H)$ NMS 실험군에서는 제 3 급아민 및 제 4 급 암모늄길항제 모두가 구조의 구별없이 질량작용의 법칙에 따라 치환되었으나 $(^3H)$ QNB 실험군에서는 제 4급 암모늄콜린 길항제들을 단일성(unity)이 아닌 높고 낮은 두 종류의 친화도를 가진 결합부위의 양상을 나타내었다. 또 비특이성 콜린길항제인 pirenzepine을 사용한 실험군에서는 두 ligands을 모두 치환시켰고 서로 다른 결합부위가 있음을 보였다. 3. Intact cell aggregates 제제를 사용한 실험에서, $(^3H)$ NMS와 $(^3H)$ QNB 모두 homogenates 제제에서와 같은 양상의 반응을 보였다. 또 $(^3H)$ NMS를 radioligand로 하여 수종의 콜린길항제와 수종의 콜린 효능제를 사용하여 약물 상호작용으로 수용체의 성질을 검토하였다. 그 결과 콜린 길항제들은 질량작용의 법칙에 따라 치환되었으나 콜린 효능제 투여군에서는 높고 낮은 두 종류의 다른 친화력의 결항부위를 나타내었다. 4. 위의 실험의 결과로,(a) 친유성콜린 길항제인 $(^3H)$ QNB는 친수성 콜린길항제인 $(^3H)$ NMS보다 훨씬 높은 결합능력을 보였으며 이것으로 수용체 특히 mAchR의 존재 장소 또는 mAchR의 형상의 일부는 세포막 표면 뿐 아니라 세포막내의 어떤 부위와도 관계가 되는 것으로 간주되는데 이것이 $(^3H)$ QNB가 $(^3H)$ NMS보다 높은 최대 결합능력 $(B_{max})$을 나타낼 이유이다. (b) 두 종류의 다른 제제에서 우리는 같은 양상의 결과를 관찰하었기에 결점이 많은 homogenates 제제보다는 intact cell aggregates 제제를 수용체 연구에 대한 새로운 실험모형(experiment model)으로 사용할 수 있는 가능성을 제시하고자 한다.

  • PDF

Retrovirus Vector를 생산하는 세포와 공동배양된 소 수정란의 E. coli LacZ 유전자 전이와 발현 (Transfer and Expression of E. coli LacZ Gene in Boving Embryos by Co-culturing with Retrovirus Vector-Producing Cells)

  • 김태완;박세필
    • 한국가축번식학회지
    • /
    • 제19권2호
    • /
    • pp.89-93
    • /
    • 1995
  • In this study was demonstrate that retrovirus-mediated gene transfer is one of the promising alternatives to the conventional pronuclear DNA microinjection approach, especially in transferring the exogenous genes into the boving embryos. By co-culturing of zona of zona-free one-cell stage embryos with the retrovirus-producing cells for 24 hours followed by 6 days of culture in virus-free medium, we could get morulae and blastocysts expressing the E. coli LacZ genes which were transferred by our retrovirus vector. The results obtained in this study are summarized as follows : 1. Addition of 5$\mu\textrm{g}$/ml of polybrene in the embryo and virus-producing cell co-culture medium did not affect development of zona-free one-cell embryo. 2. Compared with the intact embryos removal of zona at one-cell stage before co-culturing with the virus-producing cells for one day caused only slight decrease of embryo develpment. 3. Co-culture of 625 zona-free one-cell stage embryos with the virus-producing cells resulted in 65(10.4%) morulae or blastocysts, and 12.3%(8/65) of the morulae or blastocysts were E. coli LacZ positive.

  • PDF

Fluorescence Microscopy of Condensed DNA Conformations of Bacterial Cells

  • Suleymanoglu, Erhan
    • Journal of Microbiology
    • /
    • 제40권4호
    • /
    • pp.319-326
    • /
    • 2002
  • Cellular DNA in prokaryotes is organized in nucleic acid-protein self-assemblies referred to as the nucleoid. The physical forces responsible for its stability inside the poor solvent properties of the cytoplasm and their functional implications are not understood. Studies on the organisation and functioning of the cytosol of cells largely rely on experimental protocols performed in highly dilute solutions using biochemically purified molecules, which is not a reliable substitute for the situation existing in vivo. Our current research interest is focused on the characterization of biological and physical forces determining the compaction and phase separation of DNA in Escherichia coli cytoplasm. We have emphasized the effect of excluded volume in solutions with high macromolecular concentrations (macromolecular crowding) upon self-association patterns of reactions. The prokaryotic cytosol was simulated by addition of inert polymer polyethylene glycol (PEG) (average molecular weight 20000), as an agent which afterwards facilitates the self-association of macromolecules. Fluorescence microscopy was used for direct visualization of nucleoids in intact cells, after staining with DAPI (4',6-diamidino-2-phenylindole dihydrochloride). Addition of the crowding agent PEG 20,000, in increasing concentrations generated progressively enhanced nucleoid compaction, the effect being stronger in the presence of 0.2 M NaCl and 5 mM MgCl$\_$2/. Under these conditions, the nucleoids were compacted to volumes of around 2 ㎛$\^$3/ or comparable sizes with that of living cells.

투과전자현미경을 이용한 수종의 수복재가 L292세포의 미세구조 변화에 미치는 영향 (EFFECT OF SEVERAL RESTORATIVE MATERIALS ON ULRASTRUCTURAL CHANGES OF L929 CELLS USING TRANSMISSION ELECTRON MICROSCOPE)

  • 임미경
    • Restorative Dentistry and Endodontics
    • /
    • 제21권1호
    • /
    • pp.385-402
    • /
    • 1996
  • Cytotoxicity of dental restorative materials using cell culture technique has been extensively studied by various quantitative assays. The aim of this study was to investigate the microstructural change of damaged L292 cells which could not observed with light microscope. Cytotoxic effect of ZOE, Prisma APH (Densply International Inc., U.S.A.), Clearfil FII(Kuraray Co., Japan), Fuji II(GC Co., Japan) and Fuji II LC(GC Co., Japan) on cultured L292 cells were observed. Irreversible cell damage and cytolysis were found in ZOE and Fuji II groups. In Clearfil FII, mild to moderate cell damage was observed. APH group showed variable cytotoxicity. Moderate cell damage was found in Fuji II LC group. Cytotoxic effect were as follows : A condensation of the chromatin occureds along or adjacent to the inner membrane of the nuclear envelops. The nuclear envelope remained resonably intact but the contents were partially or completely lost. The cell nucleus contains clusters of markedly electron-dense interchromatin granules. The rough endoplasmic reticulum were dilated. In some mitochondira, matrix was disoriented and fused cristae were discernible. Mitochondiral swelling and woolly appearance were recognized. Large vacuoles and autolysosmes were found in cytoplasm. Some breaks of the cytoplasmic membrane and even cytolysis could be seen in dying cells.

  • PDF