Trace-stimuli-triggered controlled degradation for hydrogel adhesives

Reactive oxygen and nitrogen species in patients with rheumatoid arthritis as potential biomarkers for disease activity and the role of antioxidants

Increased Reactive Oxygen Species and the B-Cell Receptor in Chronic Lymphocytic Leukemia Signaling

Seminal reactive oxygen species and sperm motility and morphology in men with spinal cord injury

Analysis of the Metabolic State of Residual Multiple Myeloma Plasma Cells Reveals a Proliferative Subset with Glycolytic Gene Expression

Fremyella diplosiphon alters the phycobiliprotein composition of its light-harvesting complexes, i.e., phycobilisomes, and its cellular morphology in response to changes in the prevalent wavelengths of light in the external environment in a phenomenon known as complementary chromatic acclimation (CCA). The organism primarily responds to red light (RL) and green light (GL) during CCA to maximize light absorption for supporting optimal photosynthetic efficiency. Recently, we found that RL-characteristic spherical cell morphology is associated with higher levels of reactive oxygen species (ROS) compared to growth under GL where lower ROS levels and rectangular cell shape are observed. The RL-dependent association of increasedROS levels with cellular morphology was demonstrated by treating cells with a ROS-scavenging antioxidant which resulted in the observation of GL-characteristic rectangular morphology under RL. To gain additional insights into the involvement of ROS in impacting cellular morphology changes during CCA, we conducted experiments to study the temporal dynamics of changes in ROS levels and cellular morphology during transition to growth under RL or GL. Alterations in ROS levels and cell morphology were found to be correlated with each other at early stages of acclimation of low white light-grown cells to growth under high RL or cells transitioned between growth in RL and GL. These results provide further general evidence that significant RL-dependent increases in ROS levels are temporally correlated with changes in morphology toward spherical. Future studies will explore the light-dependent mechanisms by which ROS levels may be regulated and the direct impacts of ROS on the observed morphology changes.

1. Aim/Background: Reactive oxygen species (ROS) have been identified as compounds responsible for producing cellular damage. The purpose of this research is to examine if there is production of reactive oxygen species through free radical intermediates within human hepatocytes treated with morphine, bilirubin, or furosemide. The investigation examines the early stages of biotransformation by measuring the levels of reactive oxygen species produced inside of the treated hepatocytes within the first and second hours of treatment. The experiment was designed upon a case of a jaundiced (elevated bilirubin) infant who received morphine and furosemide and later died through unknown mechanisms. The experiment looks to examine if these drug compounds could contribute to cellular damage. This can help to further understand the potential interactions and complications of free radical intermediates produced during the phases of biotransformation. 2. Method: Previously cultured human hepatocytes were washed by centrifugation and re-suspended in 1x supplemental buffer to a concentration of 1x106 cells/mL and seeded in a dark clear bottom 96-well microplate at 100,000 stained cells/well. The cells were treated with either furosemide, morphine, bilirubin, a Tert-Butyl hydro peroxide (TBHP) positive control, or left as a background. Reactive oxygen generated in the presence of these agents were quantified by fluorescence excitation/emission measurement at 495nm/529nm. Fluorescence was measured at one and two hours. ROS generated convert 2',7'-dichlorodihydrofluorescein diacetate to 2',7'-dichlorodihydrofluorescein within the cells, which fluoresces. The fluorescence intensity detected is equivalent to the level of ROS generated. Wells that were untreated were used as blanks and subtracted from background and TBPH. 3. Results: Furosemide and Morphine did not produce statistically significant levels of ROS (p >0.05) above the background in both hours 1 and 2 of biotransformation and ROS measurement (Figure 1). Although Bilirubin did not produce statistically significant (p >0.05) levels of ROS above the background (Figure 2) during the first hour, it did produce statistically significant levels in the second hour of biotransformation. Each compound’s level of ROS was reduced during the second hour, signaling the removal of intermediate ROS metabolites (Figure 2). The production of ROS in each compound signifies that there is biotransformation to an intermediate that produces ROS. 4. Conclusion: The production of ROS above the background by each of the compounds shows there is an intermediate free radical compound that is produced during the biotransformation of each compound [21]. In this study, although furosemide and morphine did not produce statistically significant levels of ROS in both hours of biotransformation, bilirubin did produce significant levels of ROS in the second hour of biotransformation. This finding is in line with previous studies that shows morphine to offer protective effects against ROS production [16, 17]; and bilirubin demonstrating deleterious production of ROS at high doses [18]. Further work must be done to examine the correlation between the levels of ROS and extent of hepatocellular damage.

Confocal laser scanning microscopy analysis of bioenergetic potential and oxidative stress in fresh and frozen-thawed human ovarian tissue from oncologic patients

Effect of Antioxidant N-Acetylcysteine Treatment on the Phenotype of the TCL-1 Transgenic Mouse Model of B-CLL through Reactive Oxygen Species (ROS) Modulation

UVB Radiation Induces Apoptosis in Keratinocytes by Activating a Pathway Linked to “BLT2-Reactive Oxygen Species”

Objective: By assessing silent mating-type information regulation 2 homolog 1 (SIRT1) nucleocytoplasmic shuttling and reactive oxygen species (ROS) levels in peripheral blood mononuclear cells (PBMCs), this study aimed to explore the role of SIRT1 in premature infants after exposure to hyperoxia and assess the protective effects of resveratrol (Res). Methods: Firstly, ROS levels as well as SIRT1 translocation and expression in PBMCs samples were evaluated from 40 premature infants with different oxygen amounts received at birth. Then, PBMCs, from additional 40 premature infants administered no oxygen at birth, were used to establish an in vitro model of hyperoxia. Results: In infants that received O2 at birth, ROS and MDA levels, and SIRT1 translocation rates gradually increased in a concentration-dependent manner, while SIRT1 gradually decreased. In agreement, PBMCs cultured in vitro showed increased ROS levels after exposed to hyperoxia, SIRT1 translocation increased as well. However, treatment with Res resulted in opposite effects. Conclusion: Res inhibits ROS release in PBMCs from preterm infants exposed to hyperoxia, likely by preventing SIRT1 nucleocytoplasmic shuttling and increasing SIRT1 expression.

High grade serous carcinoma (HGSC) of the ovary constitutes the majority of incidence and death of ovarian cancer, mainly because of the elusive etiology and cell of origin. Ovulation has been regarded as a strong risk factor and use of oral contraceptives (OC) as a protective factor for ovarian cancer with obvious dose (the span of ovulation life and duration of OC use) effect, but the mechanism is unknown. Meanwhile, more and more evidences have indicated the fallopian tube fimbriae to be the tissue of origin of HGSC. We hypothesize that high level of reactive oxygen species (ROS), which is indispensible for ovulation, as well as abundant mitogens in the mature follicular fluid is responsible for the initiation (DNA double strand breaks, DSB) and promotion (proliferation) of carcinogenesis to the fallopian tube secretory cells. We first tested the scrapped cells of the human fallopian tube epithelium for the extending of DSB. By flow cytometry with gating for the secretory cell marker PAX8 and DSB marker γH2AX, we found the secretory cells in the fimbriae harbored significantly more DSB than the those in the proximal tube. In an immortalized cell line of the human fimbriae secretory cell (we named FE25), we tested the mutagenic and mitogenic effects of the ROS and antioxidant in human follicular fluids at ovulation (FF). We found a balanced activity of of ROS and antioxidants in the undiluted follicular fluid with the antioxidants to be limited, such that, upon serial dilution, the intracellular ROS level raised proportionally. This dilution-raised effect of ROS was able to induce γH2Ax accumulation in the FE25 cells. Upon treating with sub-nM level of melatonin, the most potent and major antioxidant in FF, the level of intracellular ROS and γH2Ax readily decreased. Temporally, we found a two-stage effect of FF on the survival and proliferation of FE25, with an early phase of cell death, which occurred within 12 hours after FF treatment, followed by a cell expansion phase occurring at day 2. We interpret this two-stage effect to be an immediate ROS-induced DSB and cell apoptosis followed by a mitogen-induced proliferation of the survived cells. These survived cells carried the DSB and may be clonally expanded and move on to the carcinogenic road. We conclude that an imbalance favoring the ROS activity in the FF, likely from a significant dilution of FF by an unknown pathological status (such as fimbriae inflammation or ascites), may induce an oxidative stress and DSB in the fallopian tube fimbriae. Cells that survive this stress with some carrying DSB are expanded by the rich mitogens in the same FF. When enough mutations of critical genes were accumulated in a stem cell in this secretory population, cancer lesion such as STIC can thus arise. This abstract is also presented as Poster B5. Citation Format: Hsuan-Shun Huang, Che-Fang Hsu, Taolan Zhang, Tang-Yuan Chu. Oxidants and antioxidants in the follicular fluid in initiation of fimbriae carcinogenesis. [abstract]. In: Proceedings of the AACR Special Conference on Advances in Ovarian Cancer Research: From Concept to Clinic; Sep 18-21, 2013; Miami, FL. Philadelphia (PA): AACR; Clin Cancer Res 2013;19(19 Suppl):Abstract nr PR03.

Molecularly Classified High Risk Myeloma Cells with Low ROS Levels and High Nestin Expression Have High Proliferation Index and GEP80 Score,

Purpose: Oxidative stress, which refers to the biological damage caused by free radicals produced in excess of innate antioxidant defenses, is indicated in the ocular cancer retinoblastoma (RB). Here we have analysed the differential expression of oxidative stress responsive genes in oxidant-induced RB cells, and in RB tumor tissues.Methods: The study included cultured RB cells, and four RB tumor tissues. The reactive oxygen species (ROS) levels in Y79 cells and the RB tumor induced by hydrogen peroxide were quantified by Dichlorofluorescein (DCF) fluorescence assay. We then analysed the gene expression profile of cultured RB cells induced with hydrogen peroxide (400 µM H2O2 for 8 h) by microarray analysis, and the expression of select genes were validated in Y79 cells and RB tumor tissues by real-time PCR analysis.Results: The oxidant-induced RB tumors showed an average increase in ROS levels of 44-fold compared to induced non-neoplastic donor retina. H2O2-induced RB cell line showed a 3-fold increase in ROS levels. Microarray analysis on RB cell line induced with H2O2 showed differentially regulated genes involved in cellular processes such as: oxidative stress, angiogenesis, lipid metabolism, cell proliferation, and cell signaling pathways. Several up-regulated genes such as SOD, GPX, CAT, CDC25A, CREBBP, JUN, MMP-2, iNOS, CRYAA, RXRA, ACACB and HMGCR were validated by real-time PCR. These results corroborated with the gene expression analysis in RB tumor tissues. Relating the antioxidant gene expression with the clinico-pathologic features of the tumor tissues, we found that the tumor with invasion of choroid, optic nerve and retinal pigment epithelium, had relatively higher ROS levels and minimal antioxidant gene expression, when compared with the tumor with only choroidal invasion.Conclusions: The study suggests active involvement of redox signaling pathways in the pathogenesis of RB. Consideration of oxidative stress components in the clinical management of RB patients is emphasized.

OBJECTIVES:The aim of this study is to examine meiotic spindle in oocytes along with reactive oxygen species (ROS) levels in follicular fluid of women undergoing IVF and to correlate these findings with embryo quality and pregnancy outcome.MATERIALS AND METHODS:167 women aged 25–35 years with endometriosis (Group A), polycystic ovarian syndrome (PCOS) (Group B) and tubal block (Group C) were included. Long protocol downregulation using recombinant follicular stimulating hormone was used for ovarian stimulation. Aspirated follicular fluid containing mature oocytes were analyzed for ROS levels and the oocytes were assessed for the presence of meiotic spindle using Cri-Oosight™ Polscope. Fertilization, embryo quality, endometrial assessment, and final pregnancy outcome were assessed.RESULTS:Meiotic spindles were visualized in a higher proportion of mature oocytes retrieved from women with endometriosis (66%) as compared to those with PCOS (50.5%) and tubal block (62.3%). ROS levels were also observed to be significantly less in the follicular fluid of oocytes in women with endometriosis (Group A) as compared to the other two groups (P ≤ 0.001). However, pregnancy rates were observed to be lower in Group A (32%) than Groups B (39%) and C (44%), respectively. Within each group, oocytes with spindle visualization yielded a higher number of Grade 1 embryos (P < 0.05) as well as lower ROS levels in follicular fluid (P ≤ 0.001) as compared to those where spindle could not be visualized.CONCLUSIONS:There was good correlation between spindle imaging and ROS levels as reliable predictors of oocyte assessment. Women with endometriosis had low ROS levels and good spindle imaging results suggesting a possible role of endometrial receptivity accounting for lower pregnancy rates in these women. Poor oocyte quality, as reflected by higher mean ROS levels and low number of oocytes with spindle visualization, could be the factor impeding pregnancy in women with PCOS as compared to women with tubal block.

As a malignant head and neck cancer, nasopharyngeal carcinoma (NPC) has high morbidity. Parkin expression has been reported to be reduced in NPC tissues and its upregulation could enhance paclitaxel-resistant cell cycle arrest. This study was performed to explore the possible mechanism of Parkin related to B-cell lymphoma-2 (Bcl-2)/adenovirus E1B 19 kDa interacting protein 3 (BNIP3)/BNIP3-like (NIX)-mediated mitochondrial autophagy in NPC cells. Initially, after Parkin overexpression or silencing, cell viability and proliferation were evaluated by lactate dehydrogenase and colony formation assays. JC-1 staining was used to assess the mitochondrial membrane potential. In addition, the levels of cellular reactive oxygen species (ROS) and mitochondrial ROS were detected using DCFH-DA staining and mitochondrial ROS (MitoSOX) red staining. The expression of proteins was measured using Western blot. Results showed that Parkin overexpression inhibited, whereas Parkin knockdown promoted the proliferation of paclitaxel-treated NPC cells. Besides, Parkin overexpression induced, whereas Parkin knockdown inhibited mitochondrial apoptosis in paclitaxel-treated NPC cells, as evidenced by the changes of Cytochrome C (mitochondria), Cytochrome C (cytoplasm), BAK, and Bcl-2 expression. Moreover, the levels of ROS, mitochondrial membrane potential, and LC3II/LC3I in paclitaxel-treated C666-1 cells were hugely elevated by Parkin overexpression and were all declined by Parkin knockdown in CNE-3 cells. Furthermore, Parkin upregulation activated, whereas Parkin downregulation inactivated BNIP3/NIX signaling. Further, BNIP3 silencing or overexpression reversed the impacts of Parkin upregulation or downregulation on the proliferation and mitochondrial apoptosis of paclitaxel-treated NPC cells. Particularly, Mdivi-1 (mitophagy inhibitor) or rapamycin (an activator of autophagy) exerted the same effects on NPC cells as BNIP3 silencing or overexpression, respectively. Collectively, Parkin overexpression activated BNIP3/NIX-mediated mitochondrial autophagy to enhance sensitivity to paclitaxel in NPC.