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Adaptive Response Induced by Low Dose Ionizing Raditation in Human Cervical Carcinoma Cells

  • Kim, Jeong -Hee;Lee, Kyung -Jong;Cho, Chul -Koo;Yoo, Seong -Yul;Kim, Tae -Hwan;Ji, Young -Hoon;Kim, Sung -Ho
    • Archives of Pharmacal Research
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    • v.18 no.6
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    • pp.410-414
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    • 1995
  • Adaptive response induced by low dese .gamma.-ray irradiation in human cervical carcinoma cells was examined. Cells were exposured to low dose of .gamma.-ray irradiation in human cervical carcinoma cells was examined. Cells were exposured to low dose of .gamma.-ray (1-cGy) followed by high doses of r-ray irradiation (0,1,2,3,5,7 and 9Gy for chlnogenic assay or 1.5Gy for micronucleus assay) with various time intervals. Survival fractions of cells in both low dose-irradiated and unirrated groups were analyzed by clonogenic assay. Surviva fractions of low dose-irradiated in cell survival was maximum when low and high dose irradiation time interval was 4 hr. Frequencies of micronuclei which is an indicative of chromosome aberration were also enutained from survival fractions analyzed by clonogenic assay, maximum when low and high dose irradiation time interval was 4hr. Frequencies of micronuclei which is an indicative of chromosome aberration were also enumerated in both low dose-irradiated and unirradiated groups. In consiststent with the result obtained from survival fractions analyzed by clonogenic assay, maximum reduction in frquencies of micronuclei was observed when low dose radiation was given 4 hr prior to high response to subsequent high dose .gamma.-ray irradiation in human cervical carcinomal cells. Our data suggest that one of the possible mechanisms of adaptive response induced by low dose rediation is the increase in repair of DNA double strand breaks in low dose radiation-adapted cells.

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In vivo Pharmacokinetics, Activation of MAPK Signaling and Induction of Phase II/III Drug Metabolizing Enzymes/Transporters by Cancer Chemopreventive Compound BHA in the Mice

  • Hu, Rong;Shen, Guoxiang;Yerramilli, Usha Rao;Lin, Wen;Xu, Changjiang;Nair, Sujit;Kong, Ah-Ng Tony
    • Archives of Pharmacal Research
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    • v.29 no.10
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    • pp.911-920
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    • 2006
  • Phenolic antioxidant butylated hydroxyanisole (BHA) is a commonly used food preservative with broad biological activities, including protection against chemical-induced carcinogenesis, acute toxicity of chemicals, modulation of macromolecule synthesis and immune response, induction of phase II detoxifying enzymes, as well as its undesirable potential tumor-promoting activities. Understanding the molecular basis underlying these diverse biological actions of BHA is thus of great importance. Here we studied the pharmacokinetics, activation of signaling kinases and induction of phase II/III drug metabolizing enzymes/transporter gene expression by BHA in the mice. The peak plasma concentration of BHA achieved in our current study after oral administration of 200 mg/kg BHA was around $10\;{\mu}M$. This in vivo concentration might offer some insights for the many in vitro cell culture studies on signal transduction and induction of phase II genes using similar concentrations. The oral bioavailability (F) of BHA was about 43% in the mice. In the mouse liver, BHA induced the expression of phase II genes including NQO-1, HO-1, ${\gamma}-GCS$, GST-pi and UGT 1A6, as well as some of the phase III transporter genes, such as MRP1 and Slco1b2. In addition, BHA activated distinct mitogen-activated protein kinases (MAPKs), c-Jun N-terminal kinase (JNK), extracellular signal-regulated protein kinase (ERK), as well as p38, suggesting that the MAPK pathways may play an important role in early signaling events leading to the regulation of gene expression including phase II drug metabolizing and some phase III drug transporter genes. This is the first study to demonstrate the in vivo pharmacokinetics of BHA, the in vivo activation of MAPK signaling proteins, as well as the in vivo induction of Phase II/III drug metabolizing enzymes/transporters in the mouse livers.

Optimal Conditions for the Production of Salt-tolerant Protease from Aspergillus sp. 101 and Its Characteristics (Aspergillus sp. 101로부터 내염성 단백분해효소 생산을 위한 최적 조건 및 특성)

  • Hwang, Joo-Yeon;Choi, Seung-Hwa;Lee, Si-Kyung;Kim, Sang-Moo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.38 no.11
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    • pp.1612-1617
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    • 2009
  • Aspergillus sp. 101 was isolated from the Korean traditional soybean paste for the production of a salt-tolerant protease. The optimal condition for the production of a salt-tolerant protease was determined with various energy sources such as carbon, nitrogen, and protein, and at different culture conditions such as temperature, pH, incubation time and NaCl concentration. The most favorable organic nitrogen sources were 2% defatted soybean flour (DSF) and soy protein isolate (SPI). Optimal pH and temperature were pH 6.0 and $25{\sim}27^{\circ}C$, respectively. Therefore, Aspergillus sp. 101 protease was a mild acid (or neutral) protease. Protease production was the highest at 0.1% concentration of $CaCO_3,\;K_2HPO_4$ and Arabicgum. Aspergillus sp. 101 could grow in culture medium at 15% NaCl concentration and produce a salt-tolerant protease even at 7% NaCl. The cell mass and protease activity of Aspergillus sp. 101 cultured in a modified medium was comparatively higher in Czapek dox and protease producing media. Hence, Aspergillus sp. 101 protease can be utilized in soy or fish sauce industry as a salt-tolerant protease starter.

Cellular study of replicative senescence in human periodontal ligament fibroblast using molecular biology (분자생물학을 이용하여 복제노화된 사람치주인대섬유모세포의 세포학적 연구)

  • Kim, Byung-Ock;Cho, Il-Jun;Park, Joo-Cheol;Kook, Joong-Ki;Kim, Heung-Joong;Jang, Hyun-Seon
    • Journal of Periodontal and Implant Science
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    • v.35 no.3
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    • pp.623-634
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    • 2005
  • Human periodontal ligament fibroblast(hPDLF) is very important to cure periodontal tissue because it can be diverged into various cells. This study examined the expression of MMP-1, TIMP-1, periodontal ligament specific PDLs22, Type I collagen, Fibronectin, TIMP-2, telomerase mRNA in a replicative senescence of hPDLF. The periodontal ligament tissue was obtained from periodontally healthy and non-carious human teeth extracted for orthodontic reasons at the Chosun University Hospital of Dentistry with the donors' informed consent. The hPDLF cells were cultured in a medium containing Dulbecco's modified Eagle medium(DMEM, Gibco BRL, USA) supplemented with 10% fetal bovine serum(FBS, Gibco BRL, USA) at 37C in humidified air with 5% $CO_2$. For the reverse transcription-polymerase chain reaction(RT-PCR) analysis, the total RNA of the 2, 4, 8, 16, 18, and 21 passage cells was extracted using a Trizol Reagent(Invitrogen, USA) in replicative hPDL cells. Two passage cells, i.e. young cells, served as the control, and ${\beta}-actin$ served as the internal control for RT-PCR The results of this study about cell morphology and gene expression according to aging of hPDLF using RT-PCR method are as follows: 1. The size of hPDLF was increased with aging and it was showed that the hPDLF was dying in the final passage. 2. PDLs22 mRNA was expressed in young hPDLF of the two, four, and six passage. 3. TIMP-1 mRNA was expressed in young hPDLF of the two and four passage. 4. There was a tendency that MMP-1 mRNA was weakly expressed over eighteen. 5. Type 1 collagen mRNA was expressed in almost all passages, but it was not expressed in the final passage. 6. Fibronectin mRNA was observed in all passages and it was weakly expressed in the final passage. 7. TIMP-2 and telomerase mRNA were not expressed in this study. Based on above results, it was observed that PDLs22, Type 1 collagen, Fibronectin, MMP-1. and TIMP-1 mRNA in hPDLF were expressed differently with aging. The study using the hPDLF that is collected from healthy patients and periodontitis patients needs in further study.

Pharmacological Action of Ginseng (인삼의 약리작용)

  • 홍사악;임정규;박찬웅;차인준
    • Journal of Ginseng Research
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    • v.3 no.1
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    • pp.66-93
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    • 1979
  • Panax ginseng C. A. Meyer, which has been known for more than EWO years. occupies a Particular prince in folk medicine as so called tonic remedy. The pharmacolgical investigations of ginseng, based on the scientific concepts and methodology, have been performed by many researchers through the past 50∼60 years at different parts of the world. The pharmacological action of Panax ginseng compiled from the numerous reports can be summarized as follows: 1. On central nervous system, the effect of Panax ginseng is timulatory in smaller doses and somewhat depressive in larger doses. From the psychopharmacological aspect, ginseng seems to increase the mental efficiency of man. 2. Ginseng has the effect tending to Protect organism from various physical and chemical stresses. 3. The growth and basal metabolic rates of experimental animals are stimulated by ginseng. Ginseng also prolongs the survival time of animals under adverse influences. 4. Increasing the physical and mental efficiency, ginseng postpones the onset of fatigue and increases the working capacities. 5. In the case of the intravenous administration of ginseng, a transitory and slight hypotensive effect is observed. These hypotensive effects seems to include that of a direct action and actions related to the release of histamine and/or serotonin by ginseng. 6. It is Presumed that ginseng lowers the elevated bleed ingar and cholesterol level. 7. Ginseng tends to increase the gastrointestinal motizity and tone 8. It is presumed that ginseng Promotes the iron metabolism and activates the hematopoietic factors. 9. Ginseng tends to stimulate the biosynthesis of nucleic acid and release of histamine and serotonin. 10. The toxicity end adverse reactions of ginseng appear to be nothing that warrants apprehension. 11. Anticancer erects of ginseng seem to be due to indirect action rather than direct action on cancer cell, by improving the host condition 12. Recent clinical trials of ginseng harts obtained sent good results, but Present trial is still limited in its range, so it is necessary to broaden the scope of trial covering many kinds of organs and diseases. From the above, although it appears that substantial advancements have been achieved in the studies on the Pharmacological actions of Panax ginseng there are many discrepancies noticed in the reported data. Furthermore the precise mechanisms of actions of ginseng are sometimes obscure, even unknown in other actions as the students stand now. The main reasons for this are considered to be that even though saponin has been identified at one of the active substances of ginseng, other components have not fully been identified and that the experimental approaches of the investigations varied with different researchers. Thus a thorough analysis of the chemical components and newer standardized concepts and metohds appear to be the pre-requisites for further study of the pharmacolgical effects and mechaisms of Panax ginseng.

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Culture Conditions for Hydrogen Production of Enterobacter cloacae YJ-1 (수소생산을 위한 Enterobacter cloacae YJ-1의 배양조건)

  • Lee Ki-Seok;Kang Chang-Min;Chung Seon-Yong
    • KSBB Journal
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    • v.19 no.6 s.89
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    • pp.446-450
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    • 2004
  • We investigated the effective culture conditions of anaerobic bacteria, Enterobacter cloacae YJ-1 on hydrogen production. It was cultured with 60 mL of working volume at $35^{\circ}C$, 120 rpm for 40 h. With culture time, hydrogen production and cell growth increased, but residual glucose and pH decreased. When the $2\%$ of glucose was used as single carbon source, hydrogen production was 975.1 mL/L. To enhance hydrogen productivity, mixed carbon sources of glucose and sucrose were added. The maximum hydrogen production was earned at the mixing ratio of 25:75, and it was 1319.5 mL/L. When we added 50 mM of phosphate to protect the pH drop in culture broth, hydrogen production increased 1.3 times more than that of initial concentration. The organic nitrogen sources were more effective than inorganic nitrogen for hydrogen production. Among organic nitrogen, yeast extract was the most effective and its hydrogen production was 1691.3 mL/L. Among 9 of mineral sources, Ferric citrate and $NaMoO_4$ were especially effective, and their productions were 1782.3 mL/L and 1784.8 mL/L, respectively.

A Study on the Protective Effect and Its Mechanism of Zinc against Immuno-cytotoxicity of Methylmercury (유기수은의 세포면역독성과 이에 대한 아연의 방어효과 및 기전)

  • 고대하;염정호;오경재
    • Journal of Environmental Health Sciences
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    • v.27 no.2
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    • pp.82-91
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    • 2001
  • This study was carried out to elucidate the protective effect of zinc chloride(ZnCl$_2$) and its mechanism against the immuno-cytotoxicity of methylmercury chloide($CH_3$HgCl). This study was observed in the culture of EMT-6 cells which are originated from mammary adenocarcinoma of Balb/c mouse. Cytotoxicity of metals was measured by cell viability and NO$_2$$^{[-10]}$ , and mitochondrial function was evaluated by adenosine triphosohate (ATP) production. $CH_3$HgCl significantly decreased the sythesis of nitric oxide(NO), ATP and glutathione(GSH) in a dose-dependent manner. ZnCl$_2$ significantly increased the synthesis of GSH in a dose-dependent manner, but synthesis of NO and ATP were not changed. The immuno-cytotoxicity of $CH_3$HgCl was not fully protected when combined addition of ZnCl$_2$, whereas ZnCl$_2$ prior to addition of $CH_3$HgCl completly protected the Hg-induced immuno-cytotoxicity. Similarly, intracellular accumulation of mercury significantly decreased by ZnCl$_2$. Degree of diminution of intracellular mercury was larger in ZnCl$_2$ prior to addition of $CH_3$HgCl than in combined addition of ZnCl$_2$ and $CH_3$HgCl.. Dithiothreitol(DTT) or buthionine sulfoximine(BSO) addition at 50$\mu$M or less, which was not toxic to the cells, did not affect synthesis of NO and ATP. DTT increased intracellular GSH level and DTT pretreatment protected toxicity induced by $CH_3$HgCl as shown complete recover in the NO and ATP values. BSO decreased intracellular GSH level and BSO pretreatment exaggerated toxicity induced by $CH_3$HgCl as shown synergistic reduction in the NO and ATP values. These results indicated that the protective effects of zinc against immuno-cytotoxicity of methylmercury associated with increasing cellular level of GSH. Increased intracellular GSH transports methylmercury to out of cells. In accordance with intracellular level of mercury decreased, immuno-cytotoxicity of methylmercury decreased. These result also suggest that the protective mechanism of zinc against the mercury toxicity would be exerted in the immune system in vivo.

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The effect of human mesenchymal stem cell injection on pain behavior in chronic post-ischemia pain mice

  • Yoo, Sie Hyeon;Lee, Sung Hyun;Lee, Seunghwan;Park, Jae Hong;Lee, Seunghyeon;Jin, Heecheol;Park, Hue Jung
    • The Korean Journal of Pain
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    • v.33 no.1
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    • pp.23-29
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    • 2020
  • Background: Neuropathic pain (NP) is considered a clinically incurable condition despite various treatment options due to its diverse causes and complicated disease mechanisms. Since the early 2000s, multipotent human mesenchymal stem cells (hMSCs) have been used in the treatment of NP in animal models. However, the effects of hMSC injections have not been studied in chronic post-ischemia pain (CPIP) mice models. Here, we investigated whether intrathecal (IT) and intrapaw (IP) injections of hMSCs can reduce mechanical allodynia in CPIP model mice. Methods: Seventeen CPIP C57/BL6 mice were selected and randomized into four groups: IT sham (n = 4), IT stem (n = 5), IP sham (n = 4), and IP stem (n = 4). Mice in the IT sham and IT stem groups received an injection of 5 μL saline and 2 × 104 hMSCs, respectively, while mice in the IP sham and IP stem groups received an injection of 5 μL saline and 2 × 105 hMSCs, respectively. Mechanical allodynia was assessed using von Frey filaments from pre-injection to 30 days post-injection. Glial fibrillary acidic protein (GFAP) expression in the spinal cord and dorsal root ganglia were also evaluated. Results: IT and IP injections of hMSCs improved mechanical allodynia. GFAP expression was decreased on day 25 post-injection compared with the sham group. Injections of hMSCs improved allodynia and GFAP expression was decreased compared with the sham group. Conclusions: These results suggested that hMSCs may be also another treatment modality in NP model by ischemia-reperfusion.

Fermentation Process for Mass Production of Clitocybin A, a New Anti-Wrinkle Agent from Clitocybe aurantiaca and Evaluation of Inhibitory Activity on Matrix Metalloproteinase-1 Expression (Clitocybe aurantiaca 균주가 생산하는 주름개선소재 clitocybin A의 대량 발효생산 및 MMP-1 발현저해활성)

  • Kim, Kwan-Chul;Lee, Hyeok-Won;Lee, Hong-Won;Choo, Soo-Jin;Yoo, Ick-Dong;Ha, Byung-Jo
    • Microbiology and Biotechnology Letters
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    • v.42 no.2
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    • pp.194-201
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    • 2014
  • Clitocybin A is a novel anti-wrinkle cosmetic agent produced by the strain from a Korean native mushroom Clitocybe aurantiaca. In this study, fermentation, extraction, and purification conditions for a large scale production of clitocybin A were optimized, and its cytotoxicity and inhibition activity on the expression of matrix metalloproteinase-1 (MMP-1) were characterized. The mass production of anti-wrinkle agent was achieved according to the 300 L fermentation process with a fed-batch cultivation using the modified yeast-maltose (YM) broth, and a total of 12.5 kg of cell mass was obtained in a 120 L culture broth for 14 days. After extraction and purification, clitocybin A was identified by HPLC. The cytotoxicity of clitocybin A was examined by the MTT assay. When assayed at 100 and 200 ${\mu}g/ml$ concentrations, clitocybin A showed no cytotoxicity, demonstrating safety. The inhibition activity of clitocybin A on the expression of MMP-1 was examined against UV irradiation. Oleanolic acid (control group) showed a relatively low MMP-1 inhibiting activity (ca. 16.7%) at 10 ${\mu}g/ml$ and showed increased cytotoxicity at higher concentrations. In contrast, clitocybin A showed no cytotoxicity at 100 ${\mu}g/ml$, and exhibited a relatively high MMP-1-inhibiting activity (33.1%). These findings indicate that clitocybin A may be a safe and effective anti-wrinkle agent for use in functional cosmetics.

Sex Determination in Somatic and Embryonic Cells of the Pig by FISH and PCR (FISH와 PCR에 의한 돼지 체세포 및 배아세포의 성 판정)

  • Chung, Y.;Jeon, J.T.;Kim, K.D.;Lee, S.H.;Hong, K.C.
    • Korean Journal of Animal Reproduction
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    • v.20 no.3
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    • pp.323-331
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    • 1996
  • Predetermination of sex in mammalian species has many aspects of application including the prenatal diagnoses of genetic disorders in humans and sex-selected breeding programs in the animal industry. Embryos sexing can be carried out using the polymerase chain reaction (PCR) to amplify specific sequences present in the sex chromosomes, or by fluorescent in situ hybridization (FISH) of specific probes to the X and Y chromosomes. A 3.3 kb porcine male-specific DNA fragment (pEM39) was cloned previously in our laboratory. In this study, FISH and PCR methods were employed to examine if the pEM39 can be used a sex-specific DNA probes Porcine ovaries were obtained from a local slaughter house and oocytes collected. All oocytes were subjected to in vitro maturation followed by 1n vitro fertilization. Parthenogenetically activated embryos were served as a negative control. Embryonic samples were collected at the 2-cell stages and PCR was performed to analyze DNA. Among 10 embryos examined, four embryos were identified as males and six were females. The cloned male-specific DNA fragment showed male-specificity for the cells in the liver tissue and the porcine early embryos by FISH. It was also demonstrated that the cloned male-specific DNA is localized on the hetero chromatic region of the long arm in the Y chrom-osome (Yq) as shown by the FISH and karyotyping. The results suggest that the cloned male-specific DNA fragment may be useful for predetermination of sex with a few embryonic cells. The porcine male-specific sequence can be a reliable index for embryo sexing by PCR.

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