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Promoting Effects of Sanguinarine on Apoptotic Gene Expression in Human Neuroblastoma Cells

  • Cecen, Emre;Altun, Zekiye;Ercetin, Pinar;Aktas, Safiye;Olgun, Nur
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.21
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    • pp.9445-9451
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    • 2014
  • Neuroblastoma is the most common extracranial solid tumor in children. Approximately half of the affected patients are diagnosed with high-risk poor prognosis disease, and novel therapies are needed. Sanguinarine is a benzophenanthridine alkaloid which has anti-microbial, anti-oxidant and anti-inflammatory properties. The aim of this study is whether sanguinarine has in vitro apoptotic effects and which apoptotic genes might be affected in the human neuroblastoma cell lines SH-SY5Y (N-myc negative), Kelly (N-myc positive, ALK positive), and SK-N-BE(2). Cell viability was analysed with WST-1 and apoptotic cell death rates were determined using TUNEL. After RNA isolation and cDNA conversion, expression of 84 custom array genes of apoptosis was determined. Sanguinarine caused cell death in a dose dependent manner in all neuroblastoma cell lines except SK-N-BE(2) with rates of 18% in SH-SY5Y and 21% in Kelly human neuroblastoma cells. Cisplatin caused similar apoptotic cell death rates of 16% in SH-SY5Y and 23% in Kelly cells and sanguinarine-cisplatin combinations caused the same rates (18% and 20%). Sanguinarine treatment did not affect apoptototic gene expression but decreased levels of anti-apoptotic genes NOL3 and BCL2L2 in SH-SY5Y cells. Caspase and TNF related gene expression was affected by the sanguinarine-cisplatin combination in SH-SY5Y cells. The expression of regulation of apoptotic genes were increased with sanguinarine treatment in Kelly cells. From these results, we conclude that sanguinarine is a candidate agent against neuroblastoma.

Inhibitory Constituents against Cyclooxygenases from Aralia cordata Thunb

  • Dang Nguyen Hai;Zhang XinFeng;Zheng MingShan;Son Kun Ho;Chang Hyeun Wook;Kim Hyun Pyo;Bae KiHwan;Kang Sam Sik
    • Archives of Pharmacal Research
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    • v.28 no.1
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    • pp.28-33
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    • 2005
  • Seven diterpenes, four polyacetylenes, a lipid glycerol, and two sterols were isolated from the methylene chloride fraction of the root of Aralia cordata. Their chemical structures were determined as (-)-pimara-8(14), 15-dien-19-oic acid (2), pimaric acid (3), (-)-kaur-16-en-19-oic acid (4), 17-hydroxy-ent-kaur-15-en-19-oic acid (9), $7{\alpha}$-hydroxy-(-)-pimara-8(14), 15-dien-19-oic acid (10), $16\alpha$, 17 -dihydroxy-(-)-kauran-19-oic acid (11), 16-hydroxy-17-isovaleroyloxy-ent-kauran-19­oic acid (12), falcarindiol (5), dehydrofalcarindiol (6), dehydrofalcarindiol-8-acetate (7), falcarin­diol-8-acetate (8), alpha-mono palmitin (13), stigmasterol (1), and daucosterol (14) by the spectral evidences. These compounds were tested with COX-1 and COX-2 inhibition assays. This study found that compounds 2, 4, 5, 6, 7, 8, and 10 inhibited COX-1 dependent conversion of the exogenous arachidonic acid to $PGE_2$ in a dose-dependent manner with $IC_{50}$ values of $134.2{\mu}M$, $121.6{\mu}M$, $170{\mu}M$, $50.4{\mu}M$, $11.7{\mu}M$, $99.6{\mu}M$, and $69.6{\mu}M$, respectively. But, most of these compounds weakly inhibited COX-2 dependent $PGE_2$ generation. Among them, only compound 4 showed relatively significant inhibitory activity $(IC_{50}\;:\;127.6{\mu}M)$.

Assessment on Mariner's Competency

  • Kobayashi, Hiroaki;Ishibashi, Atsushi;Nishimura, Tomohisa
    • Proceedings of the Korean Institute of Navigation and Port Research Conference
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    • 2004.08a
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    • pp.129-138
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    • 2004
  • Recently, maintenance of clean ocean is very important issue. One part of the issue strongly relates to the safe navigation of vessels. Most parts of safe navigation depend on the mariner's competencies. STCW was established to guarantee the sufficient mariner's competencies. However the code dose not indicate the each necessary competency clearly. Therefore, the understandings on STCW are not same among training institutes, and each training institute interprets them individually and executes them. As a result, it makes big differences among the institutes concerning training methods and contents and the assessment on mariner's competencies. The countries in EU have paid attention on this issue and the activities enhance the rational assessment of competencies through METNET. In order to execute the rational assessments, it is necessary to clarify the techniques being assessed. And it means the necessity to clarify the necessary techniques for safe navigation and to assess the competencies to achieve them, and then we can attain the objectives of STCW. In this paper, the necessary techniques for achievement of safe navigation are discussed and the methods of assessment of competencies are proposed. As we apply proposed assessment system, we can get the mariner's competency quantitatively and continuously through training period. Then we can know the trend of the competency that is the learning process. By clarifying the learning process of the techniques, we can decide the necessary training time to achieve the competencies. Furthermore, we can discuss the issues on conversion between training onboard and simulator used training by analysing the learning processes.

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GTP Induces S-phase Cell-cycle Arrest and Inhibits DNA Synthesis in K562 Cells But Not in Normal Human Peripheral Lymphocytes

  • Moosavi, Mohammad Amin;Yazdanparast, Razieh;Lotfi, Abbas
    • BMB Reports
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    • v.39 no.5
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    • pp.492-501
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    • 2006
  • Since differentiation therapy is one of the promising strategies for treatment of leukemia, universal efforts have been focused on finding new differentiating agents. In that respect, we used guanosine 5'-triphosphate (GTP) to study its effects on K562 cell line. GTP, at concentrations between 25-200 ${\mu}M$, inhibited proliferation (3-90%) and induced 5-78% increase in benzidine-positive cells after 6-days of treatments of K562 cells. Flow cytometric analyses of glycophorine A (GPA) showed that GTP can induce expression of this marker in more mature erythroid cells in a time- and dose-dependent manner. These effects of GTP were also accompanied with inhibition of DNA synthesis (measured by [$^3H$]-thymidine incorporation) and early S-phase cell cycle arrest by 96 h of exposure. In contrast, no detectable effects were observed when GTP administered to unstimulated human peripheral blood lymphocytes (PBL). However, GTP induced an increase in proliferation, DNA synthesis and viability of mitogen-stimulated PBL cells. In addition, growth inhibition and differentiating effects of GTP were also induced by its corresponding nucleotides GDP, GMP and guanosine (Guo). In heat-inactivated medium, where rapid degradation of GTP via extracellular nucleotidases is slow, the anti-proliferative and differentiating effects of all type of guanine nucleotides (except Guo) were significantly decreased. Moreover, adenosine, as an inhibitor of Guo transporter system, markedly reduced the GTP effects in K562 cells, suggesting that the extracellulr degradation of GTP or its final conversion to Guo may account for the mechanism of GTP effects. This view is further supported by the fact that GTP and Guo are both capable of impeding the effects of mycophenolic acid. In conclusion, our data will hopefully have important impact on pharmaceutical evaluation of guanine nucleotides for leukemia treatments.

Effects of Vitamins on the Differentiation of Preadipocytes from Hanwoo Cattle Adipose Tissues

  • Lee, H.J.;Lee, S.C.;Oh, Y.K.;Han, In K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.13 no.4
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    • pp.446-450
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    • 2000
  • The experiment was conducted to study the effects of water soluble vitamins and retinoic acid on the differentiation of preadipocyte from omental, subcutaneous, intermuscular and intramuscular adipose tissue of Hanwoo. Differentiation was assessed by the change in enzyme activity, glycerol-3 phosphate dehydrogenase in serum free cell culture system. Preadipocytes treated with biotin ($10{\mu}M$) and pantothenic acid ($100{\mu}M$) were significantly (p<0.05) less differentiated than those from the control in all adipose tissue depots except intramuscular tissue. Although there was no significance, vitamin C was shown to stimulate the adipocyte conversion in omental and subcutaneous, but not in intermuscular and intramuscular adipose tissues. Lower values of GPDH activity in intermuscular preadipocyte were interpreted to be caused by relatively higher amounts of protein. In this experiment vitamin C did not stimulate fat deposition in intramuscular adipose tissue but further experiments are needed on the role of vitamin C in preadipocyte differentiation. When treated with different levels of retinoic acids, differentiation of preadipocytes was significantly (p<0.05) reduced from the level of $0.5{\mu}g/ml$ in omental and intermuscular, from $50{\mu}g/ml$ in subcutaneous, and in intramuscular at $500{\mu}g/ml$, thus showing that intramuscular preadipocytes were least responsive to retinoic acid in differentiation. All-trans retinoic acid appeared to inhibit the differentiation in a dose dependent manner, regardless of adipose tissues type.

Role of Nitric Oxide in Pepsinogen Secretion from Rat Gastric Chief Cells

  • Sung, Dae-Suk;Seo, Dong-Wan;Choi, Don-Woong;Ahn, Seong-Hoon;Hong, Sung-Youl;Lee, Hoi-Young;Han, Jeung-Whan;Lee, Hyang-Woo
    • Biomolecules & Therapeutics
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    • v.7 no.2
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    • pp.105-111
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    • 1999
  • Nitric oxide (NO), a cellular messenger synthesized from L-arginine by NO synthase (NOS, EC.1.14.13.39), is considered to be a regulator of gastric secretion. In the present study, the role of NO in the regulation of exocrine secretion was investigated in rat gastric chief cells. Treatment of chief cells with carba-chol resulted in an increase in the arginine conversion to citrulline, the amount of $NO_{x}$, the release of pepsine-gen, and the level of cGMP Especially, carbachol-stimulated increase of arginine to citrulline transformation, the amount of $NO_{x}$, cGMP level and the release of pepsinogen were partially reduced by the natural NOS inhibitor, $N^{G}$-monomethyl-L-arginine (MMA) and $N^{G}$, $N^{G}$-dimethyl-L-arginine (DMA). Furthermore, MMA- and DMA-induced decrease of pepsinogen secretion showed dose-dependent patters. Activation of NOS is one of the early events in receptor-mediated cascade of reactions in gastric chief cells and NO, not completely, but partially mediates gastric secretion. Agonist-stimulated pepsinogen secretion in chief cells has been considered to be mediated in adenosine 3',5'-cyclic monophosphate pathway and/or guanosine 3', 5'-cyclic monophosphate (cGMP) pathway. Taken together, the above results suggest that partial decrease of exocrine secretion following treatment of NOS inhibitor may result from the inactivation of NOS and subsequent guano- late cyclase, and NO/cGMP pathway may play a pivotal role in exocrine secretion.

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Ethacrynic Acid and Citral Suppressed the All Trans Retinoid-Induced Monocyte Chemoattractant Protein-1 Production in Human Dermal Fibroblasts

  • Kim, Kwang-Mi;Noh, Min-Soo;Kim, Soo-Hyun;Park, Mi-Kyung;Lee, Hye-Ja;Kim, Soo-Youl;Lee, Chang-Hoon
    • Biomolecules & Therapeutics
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    • v.18 no.1
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    • pp.71-76
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    • 2010
  • Skin irritation caused by retinol and retinoic acid results in mild erythema called as retinoid dermatitis. To develop compounds modulating the retinoid dermatitis, we tried to establish the screening method for retinoid dermatitis. At first we examined the inflammatory cytokine profile in neonatal human dermal fibroblasts which are known to be one of main site of retinoid action. As a result, interleukin-8 (IL-8) and monocytes chemoattractant protein-1 (MCP-1) were significantly produced by all trans retinoic acid (ATRA) and all trans retinol (ATROL) in dermal fibroblasts. Especially the production of MCP-1 was more than that of IL-8. The production of MCP-1 by retinoid was dose-dependently increased, continuing up to 24 hrs. After then using ethacrynic acid (ECA) known to reduce mouse ear edema induced by ATRA, we checked whether ECA suppressed the production of MCP-1. As a result, ECA effectively suppressed the production of MCP-1 in the ATRA- or ATROL-treated-fibroblasts. These results suggested that screening method effectively reflects the in vivo anti-inflammatory activity of ECA. It was reported that citral inhibited the enzyme involved in the conversion of ATROL to ATRA. We showed that citral suppressed the production of MCP-1 in ATROL-treated fibroblasts. We expect these finding might be helpful to find useful compounds modulating the side effects of retinoid or retinoid dermatitis.

Oxidative Degradation of Phenol Using Zero-Valent Iron-Based Fenton-Like Systems (영가철 기반 펜톤 시스템을 활용한 페놀의 산화분해)

  • Kim, Hak-Hyeon;Lee, Hye-Jin;Kim, Hyung-Eun;Lee, Hongshin;Lee, Byeong-Dae;Lee, Changha
    • Journal of Soil and Groundwater Environment
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    • v.18 no.4
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    • pp.50-57
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    • 2013
  • For the last couple of decades, the Fenton (-like) systems have been extensively studied for oxidation of organic contaminants in water. Recently, zero-valent iron (ZVI) has received attention as a Fenton catalyst as well as a reducing agent capable of producing reactive oxidants from oxygen. In this study, the ZVI-based Fenton reaction was assessed for the oxidative degradation of phenol using $ZVI/O_2$, $ZVI/H_2O_2$, ZVI/Oxalate/$O_2$ and hv/ZVI/Oxalate/$O_2$ systems. Reaction parameters such as pH and reagent dose (e.g., ZVI, $H_2O_2$, and oxalate) were examined. In the presence of oxalate (ZVI/Oxalate/$O_2$ and hv/ZVI/Oxalate/$O_2$ systems), the degradation of phenol was greatly enhanced at neutral pH values. It was found that ZVI accelerates the Fenton reaction by reducing Fe(III) into Fe(II). The conversion of Fe(III) into Fe(II) by ZVI was more stimulated at acidic pH than at near-neutral pH values.

Evaluation of Toyocerin, a Probiotic Containing Bacillus toyoi Spores, on Health Status and Productivity of Weaned, Growing and Finishing Pigs

  • Kyriakis, S.C.;Georgoulakis, I.;Spais, A.;Alexopoulos, C.;Miliotis, C.C.;Kritas, S.K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.16 no.9
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    • pp.1326-1331
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    • 2003
  • The aim of the study was to assess the efficacy of Toyocerin, a probiotic containing Bacillus toyoi spores, on the health status and productivity of pigs, during nursery, growing and finishing phases. On a commercial farrow-to-finish farm in Greece, 3 experimental groups were formed, each of 72 weaned piglets. The pigs of the first group (T1 group; negative controls) received normal feed with no antimicrobials or probiotics, the pigs of the second group (T2 group) received the same type of feed but supplemented with 1.0${\times}$10$^9$, 0.5${\times}$10$^9$ and 0.2${\times}$10$^9$ spores per kg of feed at weaning, growing and finishing stage, respectively, and the pigs of the third group (T3 group) were fed with Toyocerin at the dose of 1.0${\times}$10$^9$ spores per kg of feed during the entire fattening period (weaning, growing and finishing stages). The results have shown that, compared to the controls, Toyocerin treated pigs had reduced incidence of postweaning diarrhoea (p<0.05). Enterotoxigenic strains of Escherichia coli were detected in faecal samples of 0% to 25% of pigs of the treated groups, but in 33.5% to 50% of pigs of the non-treated group (p<0.05). Over the negative controls, a significant improvement of weight gain (4.5% and 8.3% for T2 and T3 groups, respectively), and of feed conversion ratio (6.6% and 13.0% for T2 and T3 groups, respectively) was observed. The 76.5% of the carcasses of the T3 group was classified in the top three categories of the EUROP scale (S, E and U), whilst the respective figures were 47.8% for T2 group and only 10.5% for T1 group (p<0.05).

Lactobacillus plantarum ZLP001: In vitro Assessment of Antioxidant Capacity and Effect on Growth Performance and Antioxidant Status in Weaning Piglets

  • Wang, J.;Ji, H.F.;Wang, S.X.;Zhang, D.Y.;Liu, H.;Shan, D.C.;Wang, Y.M.
    • Asian-Australasian Journal of Animal Sciences
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    • v.25 no.8
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    • pp.1153-1158
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    • 2012
  • The objective of this study was to evaluate the antioxidant capacity of Lactobacillus plantarum ZLP001 and its effects on growth performance and antioxidant status in weaning piglets. The survival in hydrogen peroxide and free radical-scavenging activity of Lactobacillus plantarum ZLP001 were analysed in vitro. The Lactobacillus plantarum ZLP001 showed high viability in 1.0 mmol/L hydrogen peroxide and high scavenging ability against hydroxyl, superoxide anion, and DPPH (1,1-diphenyl-2-picrylhydrazyl) radicals which was dose dependent. Ninety-six weaning piglets were selected ($7.45{\pm}0.79kg$) and divided into three groups comprising of negative control without any supplementation, treatment group with supplemented $6.8{\times}10^7$ Lactobacillus plantarum ZLP001 CFU/g of diet, and positive control with antibiotic treatment (chlorotetracycline, 80 mg/kg diet). The results showed that Lactobacillus plantarum ZLP001 supplementation enhanced feed conversion rates in piglets compared with control (p<0.05). Supplementation of Lactobacillus plantarum ZLP001 increased the concentration of superoxide dismutase (p<0.05), glutathione peroxidase (p<0.01) and catalase in serum (p<0.10), while decreased the concentration of malondialdehyde (p<0.05). The present study implies that the strain Lactobacillus plantarum ZLP001 had high antioxidant ability and its supplementation improved the growth performance and antioxidant status of weaning piglets, so it can be considered useful to alleviate oxidative stress and increase productive performance of pigs.