Leukocytes, when subjected to phagocytic, immunologic or chemical stimuli, are known to exhibit a sequence of morphological and biochemical events which lead to the production of H₂, O₂, O₂⁻, and even OH as well as the secretion of various lysosomal lytic enzymes into the extracellular environments. It was been proposed that some antiinflammatory drugs may exert their therapeutic effects by inhibiting the production of reactive oxygen species or by abrogating the effects of reactive oxygen species ontissue components. In the present study, effects of various quenchers and antiinflammatory drugs were observed on the changes of viscosity of hyaluronate and collagen gelation by oxygen radicals generated by xanthine and xanthine oxidase. Quenching effects of antiinflammatory drugs on reactive oxygen species were also observed with gas chromatography. 1. Decrease of viscosity of hyaluronate and inhibition of collagen gelation by xanthine and xanthine oxidase were inhibited by various quenchers. 2. Several oxygen radical quenchers and antiinflammatory drugs did not affect viscosity of hyaluronate and collagen gelation. 3. Reactive ouygen species generated by xanthine and xanthine oxidase affected both viscosity of hyaluronate and collagen gelation with similar pattem. Therefore, in this study quenching effects of antiinflammatory drugs on reactive oxygen species have been examined by observing viscosity changes of hyaluronate. Sodium salicylate, acetylsalicylic acid, indomethacin and hydrocortisone affected viscosity changes of hyalumnate by xanthine and xanthine oxidase. The pattem of inhibition of hyaluronate degradation by these drugs were comparable to the inhibition produced by OH scavengers and singlet oxygen quencher. 4. To danonstrate the generation of OH ethylene was determined from methional incubated with xanthine and xanthine oxidase according to gas chromatography method. Xanthine and xanthine oxidase produced ethylene fmm methional and the production was inhibited by antiinflammatory drugs. The result obtained in this study suggest that action of antiinflammatory drugs may, to some event, be attributed to their ability to intercept reactive oxygen species in addition to inhibition of synthesis of prostaglandin.
Aspirin ; Chromatography, Gas ; Collagen ; Hydrocortisone ; Indomethacin ; Leukocytes ; Methods ; Oxygen ; Reactive Oxygen Species ; Singlet Oxygen ; Sodium Salicylate ; Therapeutic Uses ; Viscosity ; Xanthine ; Xanthine Oxidase

We wished to study the intracellular transport of adenoviruses. We constructed a novel recombinant adenovirus in which the structural protein IX was labeled with a mini-singlet oxygen generator (miniSOG). The miniSOG gene was synthesized by overlapping extension polymerase chain reaction (PCR), cloned to the pcDNA3 vector, and expressed in 293 cells. Activation of miniSOG generated sufficient numbers of singlet oxygen molecules to catalyze polymerization of diaminobenzidine into an osmiophilic reaction product resolvable by transmission electron microscopy (TEM). To construct miniSOG-labelled recombinant adenoviruses, the miniSOG gene was subcloned downstream of the IX gene in a pShuttle plasmid. Adenoviral plasmid pAd5-IXSOG was generated by homologous recombination of the modified shuttle plasmid (pShuttle-IXSOG) with the backbone plasmid (pAdeasy-1) in the BJ5183 strain of Eschericia coli. Adenovirus HAdV-5-IXSOG was rescued by transfection of 293 cells with the linearized pAd5-IXSOG. After propagation, virions were purified using the CsC1 ultracentrifugation method. Finally, HAdV-5-IXSOG in 2.0 mL with a particle titer of 6 x 1011 vp/mL was obtained. Morphology of HAdV-5-IXSOG was verified by TEM. Fusion of IX with the miniSOG gene was confirmed by PCR. In conclusion, miniSOG-labeled recombinant adenoviruses were constructed, which could be valuable tools for virus tracking by TEM.
Adenoviruses, Human ; chemistry ; genetics ; metabolism ; Arabidopsis Proteins ; chemistry ; genetics ; metabolism ; Flavoproteins ; chemistry ; genetics ; metabolism ; Humans ; Phototropins ; chemistry ; genetics ; metabolism ; Singlet Oxygen ; chemistry ; Staining and Labeling ; Transfection

Light may induce various kindsof oxygen free radicals in the eye and show a toxic effect on lens epithelial cells (LEC)as well as cataract formation. In this study, we examined the oxygen free radical injury of LEC using photosensitizer like riboflavin (RF) or rese bengal (RB) and blocking effect of antagonists on photoxicity of LEC. We cultured bovine lens epithelial cells (BLEC) to 24 well plate at the concentration of 5*10 in each well. BLEC were exposed with a illumination of 1200 lux for 2 hours under th concentration of 10, 25, 50, 75 and 100 uM of RF and 5, 10, 15, and 20 uM of RB, respectively. The pattern of BLEC death was observed with inverted light microscope and the quantitation of BLEC cytotoxicity was calculated using 3-(4, 6-dimethylthiazol-2-yl) -2, 5-diphenyl tetrazolium bromide(MTT) assay. The blocking effects of catalase, sodium azide, vitamin C, and vitamin E on BLEC phototoxicity were also evaluated. The concentration of RF inducing a prominent phototoxicity with a light of 1200 lux for 2 hours was determined to 50uM and that of RB was determined to 10uM(p<0.05). The quantiative cytotoxicity of BLEC was prominent at 24 hours dark adaptation following 2 hours illumination. In the pattern of phototoxicity of BLEfC, RF induced a cell death showing shrinkage of cell membrane and arborization of process, while RB induced a cell death showing swelling of cytoplasm. In the experiment of the scavengers` effects on phototoxicity of BLEC, catalase (3.75 U/ml) attenuated selectively a phototoxicity of BLEC induced by RF and sodium azide (imM) attenuated selectively a phototoxicity induced by RB. Conclusively, we could find hydrogen peroxide and singlet oxygen were main oxygen free radicals induced by RF and RB, respectively and the patterns of cell death induced by photosensitizer of RF and RB were cellular shrinkage and swelling, repectively.
Ascorbic Acid ; Catalase ; Cataract ; Cell Death ; Cell Membrane ; Cytoplasm ; Dark Adaptation ; Dermatitis, Phototoxic* ; Epithelial Cells* ; Free Radicals ; Hydrogen Peroxide ; Lighting ; Oxygen ; Riboflavin* ; Rose Bengal* ; Singlet Oxygen ; Sodium Azide ; Vitamin E ; Vitamins

Glycyrrhiza uralensis (or licorice) is a widely used Oriental herbal medicine from which the phenylflavonoids dehydroglyasperin C (DGC), dehydroglyasperin D (DGD), and isoangustone A (IsoA) are derived. The purpose of the present study was to evaluate the antioxidant properties of DGC, DGD, and IsoA. The three compounds showed strong ferric reducing activities and effectively scavenged DPPH, ABTS+, and singlet oxygen radicals. Among the three compounds tested, DGC showed the highest free radical scavenging capacity in human hepatoma HepG2 cells as assessed by oxidant-sensitive fluorescent dyes dichlorofluorescein diacetate and dihydroethidium bromide. In addition, all three compounds effectively suppressed lipid peroxidation in rat tissues as well as H2O2-induced ROS production in hepatoma cells. This study demonstrates that among the three phenylflavonoids isolated from licorice, DGC possesses the most potent antioxidant activity, suggesting it has protective effects against chronic diseases caused by reactive oxygen species as well as potential as an antioxidant food additive.
Animals ; Benzopyrans ; Carcinoma, Hepatocellular ; Chronic Disease ; Ethidium ; Flavonoids ; Fluorescent Dyes ; Food Additives ; Glycyrrhiza ; Glycyrrhiza uralensis ; Hep G2 Cells ; Herbal Medicine ; Humans ; Isoflavones ; Lipid Peroxidation ; Rats ; Reactive Oxygen Species ; Singlet Oxygen

Apogossypolone (ApoG2), a novel derivative of gossypol, has been shown to be a potent inhibitor of antiapoptotic Bcl-2 family proteins and to have antitumor activity in multiple types of cancer cells. Recent reports suggest that gossypol stimulates the generation of cellular reactive oxygen species (ROS) in leukemia and colorectal carcinoma cells; however, gossypol-mediated cell death in leukemia cells was reported to be ROS-independent. This study was conducted to clarify the effect of ApoG2-induced ROS on mitochondria and cell viability, and to further evaluate its utility as a treatment for nasopharyngeal carcinoma (NPC). We tested the photocytotoxicity of ApoG2 to the poorly differentiated NPC cell line CNE-2 using the ROS-generating TL/10 illumination system. The rapid ApoG2-induced cell death was partially reversed by the antioxidant N-acetyl-L-cysteine (NAC), but the ApoG2-induced reduction of mitochondrial membrane potential (MMP) was not reversed by NAC. In the presence of TL/10 illumination, ApoG2 generated massive amounts of singlet oxygen and was more effective in inhibiting cell growth than in the absence of illumination. We also determined the influence of light on the anti-proliferative activity of ApoG2 using a CNE-2-xenograft mouse model. ApoG2 under TL/10 illumination healed tumor wounds and suppressed tumor growth more effectively than ApoG2 treatment alone. These results indicate that the ApoG2-induced CNE-2 cell death is partly ROS-dependent. ApoG2 may be used with photodynamic therapy (PDT) to treat NPC.
Animals ; Antineoplastic Agents ; pharmacology ; Cell Death ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Gossypol ; analogs & derivatives ; pharmacology ; Humans ; Light ; Membrane Potential, Mitochondrial ; drug effects ; Mice ; Mice, Nude ; Nasopharyngeal Neoplasms ; metabolism ; pathology ; Neoplasm Transplantation ; Photochemotherapy ; Reactive Oxygen Species ; metabolism ; Singlet Oxygen ; metabolism ; Tumor Burden ; drug effects

Animal ; Antineoplastic Agents, Phytogenic/isolation & purification/*pharmacology ; Chlorophyll/*analogs & derivatives/isolation & purification/pharmacology/radiation effects ; Comparative Study ; Drug Screening Assays, Antitumor ; Feces/*chemistry ; Hematoporphyrin Derivative ; Hematoporphyrins/pharmacology ; Human ; Leukocytes, Mononuclear/drug effects ; Mice ; Oxygen/metabolism ; Photochemistry ; Photochemotherapy ; Radiation-Sensitizing Agents/isolation & purification/*pharmacology ; Silkworms/*metabolism ; Singlet Oxygen ; Spectrophotometry ; Structure-Activity Relationship ; Support, Non-U.S. Gov't ; Tumor Cells, Cultured/*drug effects/radiation effects

PURPOSE: The purpose of this study was to evaluate the effects of essential oil on oxidative stress, immunity, and skin condition in atopic dermatitis (AD) induced mice. METHODS: This study was a 3x3 factorial design. Factors were oil type (Lavender, Thyme, and 2:1 mixture of lavender and thyme oil [blending oil]) and treatment period (0 day, 7 days, and 21 days). The samples were 45 mice with AD and randomly assigned to nine groups of five mice per group. The dependent variables such as superoxide radical, IgE, degranulated mast cells, and epidermal thickness were measured. Data were collected from February to April in 2014. Descriptive statistics, One-way ANOVA, Two-way ANOVA, and Tukey's HSD test were performed using the SPSS WIN 20.0 program. RESULTS: Dependent variables were not statistically significantly different by the three oil types (p >.05). Essential oils such as lavender, thyme, and blending oil were all effective in reducing AD symptoms and especially 2:1 blending oil were most effective. There were statistically significant differences by the three treatment periods in all dependent variables (p <.001). There were statistically significant interactions between oil types and treatment periods in all dependent variables (p <.01). For decreasing superoxide radical, degranulated mast cells, and epidermal thickness, 2:1 mixed oil should be applied for at least 21 days. Otherwise to reduce IgE, 2:1 mixed oil should be used for at least 7 days. CONCLUSION: These findings provide bases for developing effective interventions for AD patients to manage their AD symptoms.
Animals ; Dermatitis, Atopic/chemically induced/*drug therapy/pathology ; Disease Models, Animal ; *Immunity/drug effects ; Immunoglobulin E/blood ; Lavandula/*chemistry/metabolism ; Mast Cells/cytology/metabolism ; Mice ; Oils, Volatile/chemistry/pharmacology/therapeutic use ; *Oxidative Stress/drug effects ; Picryl Chloride/toxicity ; Plant Oils/chemistry/pharmacology/*therapeutic use ; Singlet Oxygen/metabolism ; Skin/drug effects/pathology ; Thymus Plant/*chemistry/metabolism