OBJECTIVE: To investigate the mechanisms underlying ozone-induced inactivation of poliovirus type 1 (PV1). METHODS: We used cell culture, long-overlapping RT-PCR, and spot hybridization assays to verify and accurately locate the sites of action of ozone that cause PV1 inactivation. We also employed recombinant viral genome RNA infection models to confirm our observations. RESULTS: Our results indicated that ozone inactivated PV1 primarily by disrupting the 5'-non-coding region (5'-NCR) of the PV1 genome. Further study revealed that ozone specifically damaged the 80-124 nucleotide (nt) region in the 5'-NCR. Recombinant viral genome RNA infection models confirmed that PV1 lacking this region was non-infectious. CONCLUSION In this study, we not only elucidated the mechanisms by which ozone induces PV1 inactivation but also determined that the 80-124 nt region in the 5'-NCR is targeted by ozone to achieve this inactivation.
5' Untranslated Regions ; Animals ; Cercopithecus aethiops ; Genome, Viral ; drug effects ; Oxidants, Photochemical ; pharmacology ; Ozone ; pharmacology ; Poliovirus ; drug effects ; Vero Cells ; Virus Inactivation

Tooth bleaching agents contain powerful oxidizing agents, which serve as the main part of bleaching agents because of its release of effective bleaching component. It has been a hot topic whether tooth bleaching agents exert negative influence on oral health. In order to provide train of thoughts and reference for further clinical researches and treatments, this review paper focuses on bleaching agents' effects on the growth of oral microbes and the formation of biofilms.
Bacteria ; drug effects ; growth & development ; Biofilms ; drug effects ; growth & development ; Humans ; Hydrogen Peroxide ; Mouth ; microbiology ; Oral Health ; Oxidants ; pharmacology ; Tooth Bleaching ; Tooth Bleaching Agents ; pharmacology

OBJECTIVETo investigate the effects of Pyrroloquinoline quinine (PQQ) on hydrogen peroxide-induced apoptosis of Schwann cells (SCs) and its mechanism.

METHODSSCs were isolated and cultured in vitro, and identified by S-100 immunofluorescence staining. The cultured SCs were divided into control group, hydrogen peroxide-treated group, hydrogen peroxide and PQQ treated groups. The intracellular superoxide dismutase (SOD) and malondialdehyde (MDA) content was detected; the apoptotic rate of SCs induced by hydrogen peroxide was determined by flow cytometry assay. The Hoechst33342 staining was used to detect the nuclear fragmentation and apoptotic nuclear condensation of SCs; the Rhodamine123 staining was used to detect the changes of mitochondrial membrane potential in SCs, the Western blot analysis was used to detect the expression of Bcl-2 in hydrogen peroxide induced SCs.

RESULTSThe SOD activity was significantly decreased and MDA level was increased in H2O2 induced SCs (P < 0.05), after addition of PQQ, the SOD content increased and MDA content decreased (P < 0.05). Flow cytometry results showed that the early apoptotic rate was 58.8% in H2O2 induced SCs, which has significant difference compared with the control group (P < 0.05), after addition of 10, 50, 100 nmol/L PQQ, the apoptotic rates were reduced to 33.7%, 18.7%, 3.9% respectively, showing significantly different with injured group (P < 0.05). Hoechst 33342 staining showed that H2O2 induced SCs had typical morphological characteristics, such as uptake of nuclear chromatin, nuclear shrinkage, nuclear fragmentation phenomenon. The proportion of apoptotic cells after PQQ treatment reduced. Rhodamine staining results showed that the H2O2 induced mitochondrial membrane potential reduction in SCs, which was reversed by addition of PQQ. Western blot analysis showed that the expression of Bcl-2 was decreased in H2O2 induced SCs, while it increased significantly after addition of PQQ (P < 0.05).

CONCLUSIONPQQ has a protective effect on oxidative stress-induced apoptosis of SCs.

Apoptosis ; drug effects ; Benzimidazoles ; Cell Nucleus ; drug effects ; DNA Fragmentation ; Fluorescent Dyes ; Humans ; Hydrogen Peroxide ; pharmacology ; Malondialdehyde ; metabolism ; Oxidants ; pharmacology ; Oxidative Stress ; Pyrroles ; pharmacology ; Quinine ; pharmacology ; Quinolines ; pharmacology ; Schwann Cells ; cytology ; drug effects ; Superoxide Dismutase ; metabolism

Oxidative stress is implicated in male infertility and significantly higher reactive oxygen species are detected in 25% of infertile males. Although different agents of various alternative medicines, including traditional Chinese medicine, have been tried with varying success, evidence remains limited on whether and how much herbs or supplements might help increase the anti-oxidant ability of the sperm. This study examined the anti-oxidative effects of icariin, a flavonoid isolated from Herba Epimedii, on the human sperm. We prepared the FeSO4/H2O2-damaged human sperms, which were co-cultured with icariin in vitro, and then observed the changes of the sperm by employing Raman micro-spectroscopy. The results showed that Raman mapping with a 514 nm excitation laser allowed clear differentiation of the nucleus, neck, and, in particular, the mitochondria-rich middle piece of a human sperm cell. The effect of icariin on different organelles of the sperm was quantified by localized spectral Raman signatures obtained within milli-seconds, and icariin could keep the "Raman fingerprint" of the human sperm the same as the control groups, suggesting that icariin could protect the human sperm from being damaged by FeSO4/H2O2. Icariin may serve as a tonifying and replenishing agent of herbal origin for enhancing reproductive functions.
Cells, Cultured ; Drugs, Chinese Herbal ; pharmacology ; Ferrous Compounds ; pharmacology ; Flavonoids ; pharmacology ; Flow Cytometry ; Humans ; Hydrogen Peroxide ; pharmacology ; L-Lactate Dehydrogenase ; metabolism ; Male ; Malondialdehyde ; metabolism ; Oxidants ; pharmacology ; Oxidative Stress ; drug effects ; Protective Agents ; pharmacology ; Spectrum Analysis, Raman ; Spermatozoa ; cytology ; drug effects ; metabolism ; Superoxide Dismutase ; metabolism ; Time Factors

OBJECTIVE: To analyze and discuss the feasibility of rabbit carotid artery treated with decellularization and photo-oxidation. METHODS: Sixty vascular slices of rabbit carotid artery were divided into a fresh group, a cryopreservation group, a glutaraldehyde group, and a decellularization plus photo-oxidation group 15 in each group. To evaluate the physical properties of all the rabbit carotid arteries by testing heat-shrinking temperature, tensile stress and the max elongation of each group. Then by buliding subcutaneous embedding model in SD rats we evaluated the biological stability and the anti-calcification function property of the above rabbit carotid arteries, and the detection means included HE stain, atomic absorption spectrometry and Von-Kossa calcium salt stain. RESULTS: The heat-shrinking temperature, tensile stress and the max elongation in the cryopreservation group were lower or shorter than those of the other groups and the difference had statistical significance (P<0.05). Although the heat-shrinking temperature and the tensile stress in the decellularization plus photo-oxidation group were lower or shorter than those in the glutaraldehyde group (P<0.05), the max elongation in the decellularization plus photo-oxidation group was much longer than that in the glutaraldehyde group (P<0.05). The rabbit carotid artery treated with decellularization plus photo-oxidation showed lower immunogenicity and better biological stability and better anti-calcification property compared with the other groups. CONCLUSION Decellularization associated with photo-oxidation is a suitable and novel protocol for small caliber artery allograft with a diameter of less than 6 mm which is unbreakable to mechanical properties and conducive to biological stability, which has a broad prospect.
Animals ; Blood Vessel Prosthesis ; Calcinosis ; prevention & control ; Carotid Arteries ; cytology ; transplantation ; Cell Separation ; methods ; Female ; Histocytological Preparation Techniques ; Male ; Oxidants, Photochemical ; pharmacology ; Oxidation-Reduction ; Rabbits ; Rats ; Rats, Sprague-Dawley ; Transplantation, Heterologous

OBJECTIVEStudy on correlation between H2O2 scavenging system and flavonoids accumulation of Scutellaria baicalensis.

METHODThe content of baicalin and baicalein in suspension cell of S. baicalensis was determined by HPLC. The content of total flavonoids and H2O2, the activity of POD and PAL was detected by UV spectrophotometry.

RESULTThe content of total flavonoid and the activity of PAL increased significantly in 12 days after 40 degrees C, dark and PEG stress. Around 12 days after NPA, NPA +40 degrees C, 40 degrees C, NPA + dark, dark and PEG stress, the content of baicalin declined and the content of baicalein rise, the activity of POD showed an increasing trend, and level of H2O2 remain stable.

CONCLUSIONModerate environmental stress could promote the accumulation of total flavonoids in S. baicalensis, baicalin convert to baicalein by POD, and maintaining the stability of H2O2 content to avoid oxidative damage.

Anti-Infective Agents ; analysis ; metabolism ; Antioxidants ; analysis ; metabolism ; Cells, Cultured ; Darkness ; Flavanones ; analysis ; metabolism ; Flavonoids ; analysis ; metabolism ; Hot Temperature ; Hydrogen Peroxide ; analysis ; metabolism ; Oxidants ; metabolism ; Oxidative Stress ; Peroxidase ; metabolism ; Phenylalanine Ammonia-Lyase ; metabolism ; Plant Extracts ; chemistry ; Plant Roots ; chemistry ; drug effects ; enzymology ; metabolism ; Plants, Medicinal ; Polyethylene Glycols ; pharmacology ; Scutellaria baicalensis ; chemistry ; drug effects ; enzymology ; metabolism ; Stress, Physiological

Neural stem cells (NSCs) have mainly been applied to neurodegeneration in some medically intractable neurologic diseases. In this study, we established a novel NSC line and investigated the cytotoxic responses of NSCs to exogenous neurotoxicants, glutamates and reactive oxygen species (ROS). A multipotent NSC line, B2A1 cells, was established from long-term primary cultures of oligodendrocyte-enriched cells from an adult BALB/c mouse brain. B2A1 cells could be differentiated into neuronal, astrocytic and oligodendroglial lineages. The cells also expressed genotypic mRNA messages for both neural progenitor cells and differentiated neuronoglial cells. B2A1 cells treated with hydrogen peroxide and L-buthionine-(S,R)-sulfoximine underwent 30-40% cell death, while B2A1 cells treated with glutamate and kainate showed 25-35% cell death. Cytopathologic changes consisting of swollen cell bodies, loss of cytoplasmic processes, and nuclear chromatin disintegration, developed after exposure to both ROS and excitotoxic chemicals. These results suggest that B2A1 cells may be useful in the study of NSC biology and may constitute an effective neurotoxicity screening system for ROS and excitotoxic chemicals.
Animals ; Brain/*cytology ; Buthionine Sulfoximine/pharmacology ; Cell Differentiation ; Cell Line ; Cell Lineage ; Cytokines/pharmacology ; Enzyme Inhibitors/pharmacology ; Excitatory Amino Acid Agonists/pharmacology ; Glutamic Acid/pharmacology ; Humans ; Hydrogen Peroxide/pharmacology ; Intercellular Signaling Peptides and Proteins/pharmacology ; Kainic Acid/pharmacology ; Mice ; Mice, Inbred BALB C ; Multipotent Stem Cells/cytology/*drug effects/physiology ; Neuroglia/cytology/drug effects/physiology ; Neurons/cytology/*drug effects/physiology ; Neurotoxins/*pharmacology ; Oxidants/pharmacology ; Phenotype ; Reactive Oxygen Species/metabolism

Down syndrome critical region 1 (DSCR1), an oxidative stress-response gene, interacts with calcineurin and represses its phosphatase activity. Recently it was shown that hydrogen peroxide inactivates calcineurin by proteolytic cleavage. Based on these facts, we investigated whether oxidative stress affects DSCR1-mediated inactivation of calcineurin. We determined that overexpression of DSCR1 leads to increased proteolytic cleavage of calcineurin. Convertsely, knockdown of DSCR1 abolished calcineurin cleavage upon treatment with hydrogen peroxide. The PXIIXT motif in the COOH-terminus of DSCR1 is responsible for both binding and cleavage of calcineurin. The knockdown of overexpressed DSCR1 in DS fibroblast cells also abrogated calcineurin proteolysis by hydrogen peroxide. These results suggest that DSCR1 has the ability to inactivate calcineurin by inducing proteolytic cleavage of calcineurin upon oxidative stress.
Adenoviridae/genetics ; Adult ; Animals ; Calcineurin/antagonists & inhibitors/*metabolism ; Cells, Cultured ; Chromatin Immunoprecipitation ; Down Syndrome/*metabolism/pathology ; Fibroblasts/metabolism/pathology ; Humans ; Hydrogen Peroxide/pharmacology ; Immunoglobulin G/immunology ; Intracellular Signaling Peptides and Proteins/*physiology ; Male ; Mice ; Mice, Inbred ICR ; Muscle Proteins/*physiology ; Neuroblastoma/genetics/metabolism/pathology ; Neurons/cytology/metabolism ; Oxidants/pharmacology ; Oxidative Stress ; Peptide Fragments/immunology ; RNA, Messenger/genetics/metabolism ; RNA, Small Interfering/pharmacology ; Rabbits ; Reverse Transcriptase Polymerase Chain Reaction ; Skin/pathology ; Young Adult

In vitro supplementation with date seed oil (DSO) can protect spermatozoa against hydrogen peroxide (H2O2)-mediated damage and can improve sperm function, possibly owing to antioxidant properties. We tested the antioxidant effects of DSO on human sperm motility, sperm viability, reacted acrosome and lipid peroxidation assessed in vitro after H2O2-mediated oxidative damage in spermatozoa. Sixteen patients (mean age: 35 years; range: 25-45 years) referred to the Histology-Embryology Laboratory of the Medicine Faculty of Sfax for semen analysis after 12-24 months of sexual intercourse without conception were selected. After spermiogram, sperm selection by two-interface discontinuous Sill Select gradient was performed, and selected spermatozoa were used in four experimental assays: control; incubation with 100 microm H2O2; incubation with 0.1% DSO; and co-incubation with 0.1% DSO and 100 microm H2O2. Motility and viability were determined using World Health Organization criteria. Acrosome reaction and lipid peroxidation were assessed by staining with fluorescein isothiocyanate-Pisum sativum and spectrophotometric measurement of malondialdehyde, respectively. Results showed that incubation with H2O2 alone led to a significant increase in lipid peroxidation (57.83%, P<0.05) associated with a significant decrease in sperm motility, sperm viability (after 30 min and 24 h) and percentage of reacted acrosome (P<0.05). Date seed oil improved sperm motility after 24 h of incubation (P<0.05) and protected spermatozoa against the deleterious effects of H2O2 on motility, viability, acrosome reaction and lipid peroxidation. We conclude that supplementation with DSO may have a function in antioxidant protection against male infertility.
Acrosome Reaction ; drug effects ; Adult ; Arecaceae ; Cell Survival ; drug effects ; Humans ; Hydrogen Peroxide ; pharmacology ; In Vitro Techniques ; Lipid Peroxidation ; drug effects ; Male ; Middle Aged ; Oxidants ; pharmacology ; Oxidative Stress ; drug effects ; Plant Oils ; pharmacology ; Seeds ; Sperm Motility ; drug effects ; Spermatozoa ; cytology ; drug effects ; metabolism

This study evaluated the biological stability and cytotoxicity of rabbit thoracic aorta cross-linked by dye-mediated photooxidation in vivo environment of blood flow. Rabbits, whose allogenic thoracic aorta was cross-linked by dye-mediated photooxidation (DMP, n=6) and glutaraldehyde (GA, n=6), were in the DMP group and GA group, respectively; rabbits, whose homogenic abdominal aorta was orthotopic transplanted (OT, n=6), were in the OT group. Then the donor arteries were transplanted into the position of the abdominal arteries of rabbits. After operation, the animals were fed for 1 month, then each of graft arteries was removed for observation. Biological stability was evaluated through histological analysis under optical microscope. Cytotoxicity was evaluated through ratio of coverage of endothelial cell (ES) under scanning-electron microscope. The results showed that no statistically significant difference was noted between DMP group and OT group (P>0.05); however, there was a significant difference between DMP group and GA group (P<0.01). The stability index in DMP group was much higher than that in GA group. There was a statistically significant difference between DMP group and GA group (P<0.01). Ratio was much higher in DMP group than in GA group. In conclusion, the arteries cross-linked by dye-mediated photooxidation treatment appeared to have higher biological stability and lower cytotoxicity in rabbit allogenic transplation model.
Animals ; Aorta, Abdominal ; transplantation ; Aorta, Thoracic ; cytology ; drug effects ; surgery ; Bioprosthesis ; Blood Vessel Prosthesis ; Coloring Agents ; chemistry ; Cross-Linking Reagents ; Endothelial Cells ; cytology ; Female ; Male ; Oxidants, Photochemical ; pharmacology ; Oxidation-Reduction ; Photochemistry ; Rabbits ; Random Allocation