• Title/Summary/Keyword: Phenylenediamine

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Quercetin Prevents Hydrogen Peroxide-induced Necrotic and Apoptotic Cell Death in Human Colonic Epithelial Cells

  • Jung, Soon-Hee
    • Korean Journal of Clinical Laboratory Science
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    • v.43 no.4
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    • pp.161-170
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    • 2011
  • Quercetin is one of the most distributed flavonoids in the plant kingdom and occurs naturally in a wide range of fruits and vegetables. This study was undertaken to determine whether quercetin exerts beneficial effect against necrotic and apoptotic cell death induced by hydrogen peroxide ($H_2O2$) in intestinal cells using the human-derived cultured T84 colonic epithelial cell line. Necrotic cell death was induced by exposing cells to 0.5 mM $H_2O_2$ for 2 h and apoptosis was induced by incubating cells in normal culture medium for 18 h following exposure of cells to 0.5 mM $H_2O2$ for 2 h. Cell viability was evaluated by the trypan blue exclusion assay and apoptosis was assessed by Hoechst 33258 staining and flow cytometry. $H_2O_2$ induced necrotic cell death in a time and dose-dependent fashion. Both necrotic and apoptotic cell deaths were not prevented by the antioxidants N,N'-diphenyl-p-phenylenediamine(DPPD) and Trolox, whereas both cell deaths induced by the organic hydroperoxide t-butylhydroperoxide (tBHP) were prevented by DPPD, suggesting that $H_2O_2$ induces cell death through a lipid peroxidation-independent mechanism. $H_2O2$-induced necrotic death was prevented by deferoxamine and 3-aminobenzamide, while the apoptotic cell death was not affected by these agents. Quercetin prevented both necrotic and apoptotic cell deaths induced by $H_2O_2$ in a dose-dependent manner. $H_2O_2$ caused activation of poly (ADP-ribose) polmerase (PARP), which was inhibited by deferoxamine, 3-aminobenzamide, and quercetin, but not DPPD. These results indicate that quercetin inhibits both necroticand apoptotic deaths of T84 cells. The anti-necrotic effect of quercetin may be attributed to its iron chelator activity rather than a direct $H_2O_2$ scavenging capacity and antioxidant. The present study suggests that quercetin may play a therapeutic role in the treatment of human gastrointestinal diseases mediated by oxidants.

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Enhanced performance of thin-film nanocomposite RO/NWF membrane by adding ZnO nanospheres in aqueous phase during interfacial polymerization process

  • Li, Hongbin;Shi, Wenying;Su, Yuheng;Hou, Hongxiang;Du, Qiyun;Zhang, Haixia;Qin, Xiaohong
    • Membrane and Water Treatment
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    • v.8 no.3
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    • pp.225-244
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    • 2017
  • A novel thin-film nanocomposite (TFN) reverse osmosis (RO)/non-woven fabric (NWF) membrane was prepared by adding zinc oxide (ZnO) nanospheres ($30{\pm}10nm$) during the interfacial polymerization process of m-phenylenediamine (MPD) and trimesoyl chloride (TMC) on self-made polysulfone (PSF) membrane/polyester (PET) non-woven fabric support. The improved performance of TFN RO membrane was verified in terms of water contact angle (WCA), water flux, salt rejection, antifouling properties and chlorine resistance. The results showed that the WCA value of TFN RO surface had a continuous decrease with the increasing of ZnO content in MPD aqueous solution. The water flux of composite TFN RO membranes acquired a remarkable increase with a stable high solute rejection (94.5 %) in $1g{\cdot}L^{-1}$ NaCl aqueous solution under the optimized addition amount of ZnO (1 wt%). The continuous testing of membrane separation performance after the immersion in sodium hypochlorite solution indicated that the introduction of ZnO nanospheres also dramatically enhanced the antifouling properties and the chlorine resistance of composite RO membranes.

Chemopreventive Activity of Turmeric Essential Oil and Possible Mechanisms of Action

  • Liju, Vijayasteltar Belsamma;Jeena, Kottarapat;Kuttan, Ramadasan
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.16
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    • pp.6575-6580
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    • 2014
  • This study aimed to evaluate the antimutagenic and anticarcinogenic activity of turmeric essential oil as well as to establish biochemical mechanisms of action. Antimutagenicity testing was accomplished using strains and known mutagens with and without microsomal activation. Anticarcinogenic activity was assessed by topical application of 7, 12 - dimethylbenz[a]anthracene (DMBA) as initiator and 1% croton oil as promoter for the induction of skin papillomas in mice. Inhibition of p450 enzymes by TEO was studied using various resorufins and aminopyrene as substrate. Turmeric essential oil (TEO) showed significant antimutagenic activity (p<0.001) against direct acting mutagens such as sodium azide ($NaN_3$), 4-nitro-O-phenylenediamine (NPD) and N-methyl-N-nitro N'nitrosoguanine (MNNG). TEO was found to have significant antimutagenic effect (>90%) against mutagen needing metabolic activation such as 2-acetamidoflourene (2-AAF). The study also revealed that TEO significantly inhibited (p<0.001) the mutagenicity induced by tobacco extract to Salmonella TA 102 strain. DMBA and croton oil induced papilloma development in mice was found to be delayed and prevented significantly by TEO application. Moreover TEO significantly (P<0.001) inhibited isoforms of cytochrome p450 (CYP1A1, CYP1A2, CYP2B1/2, CYP2A, CYP2B and CYP3A) enzymes in vitro, which are involved in the activation of carcinogens. Results indicated that TEO is antimutagenic and anticarcinogenic and inhibition of enzymes (p450) involved in the activation of carcinogen is one of its mechanisms of action.

Municipal wastewater reclamation for non-potable use using hollow- fiber membranes

  • Waghmare, Sujata;Masid, Smita;Rao, A. Prakash;Roy, Paramita;Reddy, A.V.R.;Nandy, T.;Rao, N.N.
    • Membrane and Water Treatment
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    • v.1 no.3
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    • pp.207-214
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    • 2010
  • Approximately 80% of water used in urban areas reappears as municipal wastewater (MWW). Reclamation of MWW is an attractive proposition under the present scenario of water stressed cities in India. In this paper, we attempted to reclaim MWW using lab-scale hollow- fiber (HF) membrane modules for possible reuse in non-potable applications. Experiments were conducted to evaluate the efficiency of virgin HF ($M_1$) and modified HF ($M_2$) modules. The $M_2$ module consists of HF modified with a skin layer formed through interfacial polymerization of m-phenylenediamine with trimesoyl chloride (MPD-TMC). The molecular weight cut-off (MWCO) of $M_1$ was 44000 g/mol and that of $M_2$ 10000 -14000 g/mol on the basis of rejection of polyethylene glycol. The combination of $M_1$ and $M_2$ modules was able to reduce concentrations of most of the pollutants in sewage and improved the treated water quality to the acceptable limits for non potable reuse applications. It is found that about 98-99% of the initial flux is recovered by the backwashing process, which was approximately two times in a month when operated continuously.

Use of the enzyme-linked immunosorbent assay for the detection of toxoplasmosis in swine (ELISA를 이용한 돼지 톡소플라스마병의 조기 진단에 관한 연구)

  • Suh, Myung-deuk;Jang, Dong-hwa;Joo, Hoo-don
    • Korean Journal of Veterinary Research
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    • v.29 no.4
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    • pp.567-575
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    • 1989
  • This study was conducted to evaluate the possibility of application of a microenzyme-linked immunosorbent assay(micro-ELISA) for the serodiagnosis of specific toxoplasma antibodies in swine sera and this test was performed as a microplate system by coating the polystyrene plates with toxoplasma soluble antigen, incubated serially diluted sera, then added horse radish peroxidase labelled goat anti-swine IgG(r) conjugate followed by o-phenylenediamine as substrate. The color development by enzyme-substrate reaction was determined by the photometric reading [ELISA reader at 490nm (OD)] and visual reading. The soluble antigen was prepared from the tachyzoites in mouse peritoneal cavity. A total of 1,200 swine sera from pig slaughter-house and a total of 116 swine sera from pig breeding station (S-C farm) were tested for the detection of antibodies to Toxoplasma gondii. The results obtained were summarized as follows: 1. The optimal reactions of indirect ELISA for the test sera were determined by the dilution of antigen 1:256 and 1:3,200 of horse radish peroxidase conjugate [anti-swine IgG(r)]. 2. The specific togoplasma antibody(IgG) in pigs infected with Tp artificially were detected as the serum titers of 1:64 or 1:128 at one week postinfection. 3. Of a total of 1,200 swine sera from pig slaughter-house 505 samples of sera were detected as positive (42.1%) and of a total of 116 swine sera from S-C pig breeding station 68 samples of sera as positive (58.6%). 4. The specific antibody(IgG) detection rates against a total of 1,200 test sera from pig slaughter-house were not significant between male (43.1%) and female (40.7%). 5. The indirect ELISA was proved to be a sensitive and specific procedure for the serodiagnosis of swine toxoplasmosis and also evaluated as an effective screening test for the large scale of test samples in laboratory.

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Studies on avian infectious bronchitis: II. Standardization of an indirect enzyme-linked immunosorbent assay (ELISA) for antibody measurement (닭 전염성 기관지염에 관한 연구: II. 간접 enzyme-linked immunosorbent assay(ELISA)에 의한 항체가 측정)

  • Chang, Chong-ho;Kim, Sun-joong
    • Korean Journal of Veterinary Research
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    • v.29 no.4
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    • pp.503-515
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    • 1989
  • Critical parameters affecting sensitivity and specificity of an enzyme-linked immunosorbent assay(ELISA) for detection of antibodies to avian infections bronchitis virus(IBV) were standardized. By adopting the optimized conditions an equation calculating ELISA antibody titers from the observations at single serum dilution was formulated. The purified antigen of IBV Mass-41 strain was dispensed into polystyrene microplate wells at a concentration of 300ng per well($100{\mu}l$) and the plates were coated by completey drying at $37^{\circ}C$. Diluted chicken serum and horseradish peroxidase conjugated goat anti-chicken IgG were added in order in $100{\mu}l$ volumes per well and allowed to react for 30 minutes each at room temperature. Just before use and after each reaction the plates were washed three times with distilled water. Finally o-phenylenediamine solution was added as an enzyme substrate. After incubation for another 15 minutes at room temperature absorbances were read at 492nm. Hyperimmune serum against Mass-41 strain was used as internal reference positive(IRP) serum. After repeated titration of IRP and negative serum, a constant titer of IRP was determined. Serum titrations were carried out for various sample sera together with IRP and negative sera and the observed titers of sample sera were corrected by reflecting the ratio between observed and constant titers of IRP serum. These corrected titers of the sample sera were plotted against sample/positive(S/P) OD ratios. All the OD's measured in the serum titrations were also corrected by substracting negative serum OD. The following equation was formulated from the above data; $Log_{10}$ ELISA titer=$5.568({\log}_{10}S/P)+4.161$ Thus it was possible to calculate ELISA titer by measuring absorbance at 1/400 single serum dilution. Titer measured by cross ELISA tests employing Mass-41 strain and three local IBV isolates were similar. These results suggest that the ELISA tests standardized in this study can be used for evaluating not only vaccinal immunity but also for infection status against fields IBV's.

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Role of Lipid Peroxidation on $H_2O$$_2$-Induced Renal Cell Death in Cultured Cells and Freshly Isolated Cells

  • Jung, Soon-Hee
    • Biomedical Science Letters
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    • v.8 no.4
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    • pp.251-256
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    • 2002
  • This study was undertaken to determine the underlying mechanisms of reactive oxygen species-induced cell injury in renal epithelial cells and whether there is a difference in the role of lipid peroxidation between freshly isolated renal cells and cultured renal cells. Rabbit renal cortical slices were used as a model of freshly isolated cells and opossum kidney (OK) cells as a model of cultured cells. Cell injury was estimated by measuring lactate dehydrogenase (LDH) release in renal cortical slices and frypan blue exclusion in OK cells. $H_2O$$_2$ was used as a drug model of reactive oxygen species. $H_2O$$_2$ induced cell injury in a dose-dependent manner in both cell types. However, renal cortical slices were resistant to $H_2O$$_2$ approximately 50-fold than OK cells. $H_2O$$_2$-induced cell injury was prevented by thiols (glutathione and dithiothreitol) and iron chelators (deferoxamine and phenanthroline) in both cell types. $H_2O$$_2$-induced cell injury in renal cortical slices was completely prevented by antioxidants N,N-diphenyl-p -phenylenediamine and Trolox, but the cell injury was not affected by these antioxidants in OK cells. $H_2O$$_2$ increased lipid peroxidation in both cell types, which was completely inhibited by the antioxidants. These results suggest that $H_2O$$_2$ induces cell injury through a lipid peroxidation-dependent mechanism in freshly isolated renal cells, but via a mechanism independent of lipid peronidation in cultured cells.

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Isolation and Characterization of Lactic Acid Bacteria Producing Antimutagenic Substance from Korean Dongchimi (동치미로부터 항돌연변이 물질을 생산하는 유산균의 분리 및 특성)

  • 주길재;이창호;우철주
    • Journal of Life Science
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    • v.11 no.5
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    • pp.432-438
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    • 2001
  • Various lactic acid bacteria were isolated from Korean Dongchimi (whole radish Kimichi with added water) in order to study their antimutagenic activity. Ames test using Salmonella enterica serovar typhimurium TA98 and TA100 showed the strain DLAB19 to have the highest antimutagenic activity among the 300 isolated strains against MNNG(N-methyl-N-nitro-N-nitrosoguanidine), NPD (4-nitro-O-phenylenediamine), 4-NQO(4-nitroquinoline-1-oxide) and AFB$_{1}$(aflatoxin B$_{1}$). The strain was identified as Leuconostoc mesenteroides subsp. cremoris according to the Bergeys Mannual Systematic Bscteriology based on its morphological, cultural, physiological characteristics and biological system Antimutagenic activity of Leu. mesenteroides subsp. cremoris DLAB19 was found in the culture supernatant suggesting the bacterium secretes, the antimutagenic substance in the media. The antimutagenic activity of Leu. mesenteroides subsp. cremoris DLAB19 was reconfirmed by the spore-rec assay using spores of Bacillus subtilis H17 (Rec$^{+}$) and M45 (Rec$^{[-10]}$ ).).

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Antimutagenic Effects and Isolation of Flavonoids from Humulus japonicus Extract (환삼덩굴(Humulus japonicus) 추출물의 항돌연변이 효과와 Flavonoid 성분의 분리)

  • Park, Seung-Woo;Kim, Sung-Hwan;Chung, Shin-Kyo
    • Korean Journal of Food Science and Technology
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    • v.27 no.6
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    • pp.897-901
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    • 1995
  • To investigate the physiological activities of Humulus japonicus, we examined the mutagenic and antimutagenic effects of the extract and isolated the flavonoid compounds. The water and methanol extracts of Humulus japonicus did not show the mutagenicity by Ames test in the 0.5-4 mg/plate. The methanol extract showed the antimutagenic effect aganinst Salmonella typhimurium TA98 and TA100 induced by 4-nitro-o-phenylenediamine(NPD) and 4-nitroquinoline-N-oxide(NQO). The hexane fraction showed comparatively higher antimutagenic effect than other solvent fractions from the methanol extract. The 50% inhibition concentration($IC_{50}$) of hexane fraction were 2.0 mg/plate, 0.5 mg/plate in TA98 and TA100 respectively. Quercitrin and luteolin were identified in the ethyl acetate fraction from methanol extract of Humulus japonicus by TLC and HPLC.

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Synthesis of the Low-Hygroscopic Polyimide for 2-Layer Flexible Copper Clad Laminate (2층 연성동박적층판용 저흡습 폴리이미드의 합성)

  • Kim, W.;Park, S.J.;Baek, J.O.;Gong, H.J.;Ahn, B.H.
    • Elastomers and Composites
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    • v.43 no.2
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    • pp.82-87
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    • 2008
  • In this study, nine kinds of polyimides were synthesized from 1,2,4,5-benzenetetracarboxylic dianhydride (PMDA), 4,4'-(4,4'-isopropylidenediphenoxy)bis(phthalic anhydride) (BPADA), m-pheny lenediamine (m-PDA) and 4,4'-oxydianiline (ODA) by controlling molar ratio of monomers. Synthesized polyimides were used as insulator films for 2-layer Flexible Copper Clad Laminate(FCCL) which were manufactured by the casting method. Glass transition temperature and thermal degradation temperature for 5% weight loss of the polyimide film were improved by increasing contents of m-PDA and PMDA, respectively. Water absorption of polyimide film was reduced by increasing contents of ODA and BPADA which have relatively long structure, respectively. Peel strength of 2-layer FCCL was improved by increasing contents of ODA and BPADA.