Catalytic Ferrous Iron Research Articles

Erastin induces ferroptosis via ferroportin-mediated iron accumulation in endometriosis.

Could erastin activate ferroptosis to regress endometriotic lesions? Erastin could induce ferroptosis to regress endometriotic lesions in endometriosis. Ectopic endometrial stromal cells (EESCs) are in an iron overloading microenvironment and tend to be more sensitive to oxidative damage. The feature of erastin-induced ferroptosis is iron-dependent accumulation of lethal lipid reactive oxygen species (ROS). Eleven patients without endometriosis and 21 patients with endometriosis were recruited in this study. Primary normal and ectopic endometrial stromal cells were isolated, cultured and subjected to various treatments. The in vivo study involved 10 C57BL/6 female mice to establish the model of endometriosis. The markers of ferroptosis were assessed by cell viability, lipid peroxidation level and morphological changes. The cell viability was measured by colorimetric method, lipid peroxidation levels were measured by flow cytometry, and morphological changes were observed by transmission electron microscopy. Immunohistochemistry and western blot were used to detect ferroportin (FPN) expression. Prussian blue staining and immunofluorescent microscopy of catalytic ferrous iron were semi-quantified the levels of iron. Adenovirus-mediated overexpression and siRNA-mediated knockdown were used to investigate the role of FPN on erastin-induced ferroptosis in EESCs. EESCs were more susceptible to erastin treatment, compared to normal endometrial stromal cells (NESCs) (P<0.05). Treatment of cultured EESCs with erastin dramatically increased the total ROS level (P<0.05, versus control), lipid ROS level (P<0.05, versus NESCs) and intracellular iron level (P<0.05, versus NESCs). The cytotoxicity of erastin could be attenuated by iron chelator, deferoxamine (DFO), and ferroptosis inhibitors, ferrostatin-1 and liproxstatin-1, (P<0.05, versus erastin) in EESCs. In EESCs with erastin treatment, shorter and condensed mitochondria were observed by electron microscopy. These findings together suggest that erastin is capable to induce EESC death by ferroptosis. However, the influence of erastin on NESCs was slight. The process of erastin-induced ferroptosis in EESCs accompanied iron accumulation and decreased FPN expression. The overexpression of FPN ablated erastin-induced ferroptosis in EESCs. In addition, knockdown of FPN accelerated erastin-induced ferroptosis in EESCs. In a mouse model of endometriosis, we found ectopic lesions were regressed after erastin administration. N/A. This study was mainly conducted in primary human endometrial stromal cells. Therefore, the function of FPN in vivo need to be further investigated. Our findings reveal that erastin may serve as a potential therapeutic treatment for endometriosis. This research did not receive any specific grant from funding agencies in the public, commercial or not-for-profit sectors. The authors declare no conflict of interest.

Contrasting intra- and extracellular distribution of catalytic ferrous iron in ovalbumin-induced peritonitis

Iron is an essential nutrient for every type of life on earth. However, excess iron is cytotoxic and can lead to an increased cancer risk in humans. Catalytic ferrous iron [Fe(II)] is an initiator of the Fenton reaction, which causes oxidative stress by generating hydroxyl radicals. Recently, it became possible to localize catalytic Fe(II) in situ with a turn-on fluorescent probe, RhoNox-1. Here, we screened each organ/cell of rats to globally evaluate the distribution of catalytic Fe(II) and found that eosinophils showed the highest abundance. In various cells, lysosomes were the major organelle, sharing ∼40–80% of RhoNox-1 fluorescence. We then used an ovalbumin-induced allergic peritonitis model to study the dynamics of catalytic Fe(II). Peritoneal lavage revealed that the total iron contents per cell were significantly decreased, whereas an increase in the number of inflammatory cells (macrophages, neutrophils, eosinophils and lymphocytes) resulted in an increased total iron content of the peritoneal inflammatory cells. Notably, macrophages, eosinophils and neutrophils exhibited significantly increased catalytic Fe(II) with increased DMT1 expression and decreased ferritin expression, though catalytic Fe(II) was significantly decreased in the peritoneal lavage fluid. In conclusion, catalytic Fe(II) in situ more directly reflects cellular activity and the accompanying pathology than total iron does.

Role of hemoglobin and transferrin in multi-wall carbon nanotube-induced mesothelial injury and carcinogenesis.

Multi‐wall carbon nanotubes (MWCNT) are a form of flexible fibrous nanomaterial with high electrical and thermal conductivity. However, 50‐nm MWCNT in diameter causes malignant mesothelioma (MM) in rodents and, thus, the International Agency of Research on Cancer has designated them as a possible human carcinogen. Little is known about the molecular mechanism through which MWCNT causes MM. To elucidate the carcinogenic mechanisms of MWCNT in mesothelial cells, we used a variety of lysates to comprehensively identify proteins specifically adsorbed on pristine MWCNT of different diameters (50 nm, NT50; 100 nm, NT100; 150 nm, NT150; and 15 nm/tangled, NTtngl) using mass spectrometry. We identified >400 proteins, which included hemoglobin, histone, transferrin and various proteins associated with oxidative stress, among which we selected hemoglobin and transferrin for coating MWCNT to further evaluate cytotoxicity, wound healing, intracellular catalytic ferrous iron and oxidative stress in rat peritoneal mesothelial cells (RPMC). Cytotoxicity to RPMC was observed with pristine NT50 but not with NTtngl. Coating NT50 with hemoglobin or transferrin significantly aggravated cytotoxicity to RPMC, with an increase in cellular catalytic ferrous iron and DNA damage also observed. Knockdown of transferrin receptor with ferristatin II decreased not only NT50 uptake but also cellular catalytic ferrous iron. Our results suggest that adsorption of hemoglobin and transferrin on the surface of NT50 play a role in causing mesothelial iron overload, contributing to oxidative damage and possibly subsequent carcinogenesis in mesothelial cells. Uptake of NT50 at least partially depends on transferrin receptor 1. Modifications of NT50 surface may decrease this human risk.

Ovarian endometriosis-associated stromal cells reveal persistently high affinity for iron

Ovarian endometriosis is a recognized risk for infertility and epithelial ovarian cancer, presumably due to iron overload resulting from repeated hemorrhage. To find a clue for early detection and prevention of ovarian endometriosis-associated cancer, it is mandatory to evaluate catalytic (labile) ferrous iron (catalytic Fe(II)) and to study iron manipulation in ovarian endometriotic lesions. By the use of tissues from women of ovarian endometriosis as well as endometrial tissue from women with and without endometriosis, we for the first time performed histological analysis and cellular detection of catalytic Fe(II) with a specific fluorescent probe (HMRhoNox-M), and further evaluated iron transport proteins in the human specimens and in co-culture experiments using immortalized human eutopic/ectopic endometrial stromal cells (ESCs) in the presence or absence of epithelial cells (EpCs). The amounts of catalytic Fe(II) were higher in ectopic endometrial stromal cells (ecESCs) than in normal eutopic endometrial stromal cells (n-euESCs) both in the tissues and in the corresponding immortalized ESCs. ecESCs exhibited higher transferrin receptor 1 expression both in vivo and in vitro and lower ferroportin expression in vivo than n-euESCs, leading to sustained iron uptake. In co-culture experiments of ESCs with iron-loaded EpCs, ecESCs received catalytic ferrous iron from EpCs, but n-euESCs did not. These data suggest that ecESC play a protective role for cancer-target epithelial cells by collecting excess iron, and that these characteristics are retained in the immortalized ecESCs.

Removal of polycyclic aromatic hydrocarbons from sediments using sodium persulfate activated by temperature and nanoscale zero-valent iron

The oxidation of 16 polycyclic aromatic hydrocarbons (PAHs) compounds in sediments by sodium persulfate (Na2S2O8) simultaneously activated by temperature and nano zero-valent iron (nZVI) as the source of catalytic ferrous iron was investigated. The effect of various controlling factors, including S2O82− (0.017–170 g/L), nZVI (0.01–1 g/L), and temperature (50–70 °C) were performed. The efficiency to remove PAHs was 10.7–39.1% for unactivated persulfate. The treated sample had over 50% of the persulfate still remaining from an initial persulfate dose of 170 g/L, whereas less than 1% of the persulfate remained from an initial persulfate dose of 0.017, 0.17, and 1.7 g/L. Adequate persulfate (170 g/L) must be present because it is the source of the sulfate radicals responsible for the degradation of PAHs. Results indicated that increasing temperature and the addition of nZVI into a persulfate-slurry system could enhance the persulfate oxidation process. The best removal efficiency (90%) was achieved after 24 hr while adding nZVI (0.01 g/L) to persulfate (170 g/L) at temperature of 70 °C. The results suggested that nZVI assisted persulfate oxidation without elevating temperature may be a suitable and economic alternative for the ex situ treatment of PAH-contaminated sediments.Implications: Nano zero-valent iron (nZVI) has been successfully applied to transform/degrade contaminants in soils and water. Additionally, nZVI has been used as a catalyst to activate persulfate for the treatment of various contaminants. In this study, with the support of temperature, nZVI-persulfate oxidation system for treatment of PAH-contaminated sediments was improved significantly and the treated sediment could meet remediation goals.

Catalytic ferrous iron in amniotic fluid as a predictive marker of human maternal-fetal disorders.

Amniotic fluid contains numerous biomolecules derived from fetus and mother, thus providing precious information on pregnancy. Here, we evaluated oxidative stress of human amniotic fluid and measured the concentration of catalytic Fe(II). Amniotic fluid samples were collected with consent from a total of 89 subjects in Nagoya University Hospital, under necessary medical interventions: normal pregnancy at term, normal pregnancy at the 2nd trimester, preterm delivery with maternal disorders but without fetal disorders, congenital diaphragmatic hernia, fetal growth restriction, pregnancy-induced hypertension, gestational diabetes mellitus, Down syndrome and trisomy 18. Catalytic Fe(II) and oxidative stress markers (8-hydroxy-2'-deoxyguanosine, 8-OHdG; dityrosine) were determined with RhoNox-1 and specific antibodies, respectively, using plate assays. Levels of 8-OHdG and dityrosine were higher in the 3rd trimester compared with the 2nd trimester in normal subjects, and the abnormal groups generally showed lower levels than the controls, thus suggesting that they represent fetal metabolic activities. In contrast, catalytic Fe(II) was higher in the 2nd trimester than the 3rd trimester in the normal subjects, and overall the abnormal groups showed higher levels than the controls, suggesting that high catalytic Fe(II) at late gestation reflects fetal pathologic alterations. Notably, products of H2O2 and catalytic Fe(II) remained almost constant in amniotic fluid.

Remediation of Marine Sediments Contaminated with PAHs Using Sodium Persulfate Activated by Temperature and Nanoscale Zero Valent Iron

The oxidation of 16 polycyclic aromatic hydrocarbons compounds (PAHs) in sediments by sodium persulfate (Na2S2O8) activated by temperature and nanoscale zero−valent iron (nZVI) as the source of catalytic ferrous iron was investigated. The effect of various controlling factors including S2O82− (0.017–170 g/L), nZVI (0.01–1 g/L), and temperature (50–70°C) were performed. The efficiency to remove PAHs was not too high as 10.7–39.1% for unactivated persulfate. Results from experiments indicate that increasing temperature or the addition of nZVI into a persulfate-slurry system could enhance the persulfate oxidation process. The best removal efficiency (86.3%) was attained after 24 hr while adding nZVI (0.5 g/L) to persulfate (170 g/L) at temperature of 25°C. The results of our study suggest that nZVI assisted persulfate oxidation without elevating temperature is a suitable and economic alternative for the ex–situ treatment of PAHs contaminated sediments.

Histological detection of catalytic ferrous iron with the selective turn-on fluorescent probe RhoNox-1 in a Fenton reaction-based rat renal carcinogenesis model

Iron overload of a chronic nature has been associated with a wide variety of human diseases, including infection, carcinogenesis, and atherosclerosis. Recently, a highly specific turn-on fluorescent probe (RhoNox-1) specific to labile ferrous iron [Fe(II)], but not to labile ferric iron [Fe(III)], was developed. The evaluation of Fe(II) is more important than Fe(III) in vivo in that Fe(II) is an initiating component of the Fenton reaction. In this study, we applied this probe to frozen sections of an established Fenton reaction-based rat renal carcinogenesis model with an iron chelate, ferric nitrilotriacetate (Fe-NTA), in which catalytic iron induces the Fenton reaction specifically in the renal proximal tubules, presumably after iron reduction. Notably, this probe reacted with Fe(II) but with neither Fe(II)-NTA, Fe(III) nor Fe(III)-NTA in vitro. Prominent red fluorescent color was explicitly observed in and around the lumina of renal proximal tubules 1 h after an intraperitoneal injection of 10–35 mg iron/kg Fe-NTA, which was dose-dependent, according to semiquantitative analysis. The RhoNox-1 signal colocalized with the generation of hydroxyl radicals, as detected by hydroxyphenyl fluorescein (HPF). The results demonstrate the transformation of Fe(III)-NTA to Fe(II) in vivo in the Fe-NTA-induced renal carcinogenesis model. Therefore, this probe would be useful for localizing catalytic Fe(II) in studies using tissues.

Immunohistochemical localization of bilirubin oxidation in human placenta

Immunohistochemical localization of bilirubin oxidation in human placenta

Fenton and Photo-Fenton Oxidation of Petroleum Aromatic Hydrocarbons Using Nanoscale Zero-Valent Iron

AbstractThis study investigated the feasibility of nanoscale zero-valent iron(nZVI) as the source of catalytic ferrous iron for Fenton and photo-Fenton oxidation of petroleum aromatic hydrocarbons. The oxidation processes were able to degrade more than 99% of benzene, toluene, ethylbenzene, and xylene (BTEX) within 15 min at pH 3.0, which the initial concentrations of BTEX, nZVI, and H2O2 were 4,000 μg/L, 9 mM, and 150 mg/L, respectively. The average pseudo-first-order constants for BTEX degradation were 0.309 and 0.368 min−1 at pH 3.0 for Fenton and photo-Fenton processes, respectively. Experiments revealed that by increasing pH, solubility of nZVI decreased significantly, while ultraviolet (UV) irradiation improved nZVI solubility and subsequently BTEX degradation. Cations and anions were found to reduce oxidation efficiencies of BTEX; among anions, SO42− and CO32− had the most negative effect on the removal efficiencies. On the other hand, Mg2+ and Ca2+ played a considerable inhibitor role among cat...

폐영가철 투수성반응벽체를 이용한 Modified Fenton 산화에 의한 MTBE 처리연구

MTBE (Methyl tertiary-butyl ether) has been commonly used as an octane enhancer to replace tetraethyl lead in gasoline, because MTBE increases the efficiency of combustion and decreases the emission of carbon monoxide. However, MTBE has been found in groundwater from the fuel spills and leaks in the UST (Underground Storage Tank). Fenton's oxidation, an advanced oxidation catalyzed with ferrous iron, is successful in removing MTBE in groundwater. However, Fenton's oxidation requires the continuous addition of dissolved Fe 2+ . Zero-valent iron is available as a source of catalytic ferrous iron of MFO (Modified Fenton's Oxidation) and has been studied for use in PRBs (Permeable Reactive Barriers) as a reactive material. Therefore, this study investigated the condition of optimization in MFO-PRBs using waste zero-valent iron (ZVI) with the waste steel scrap to treat MTBE contaminated groundwater. Batch tests were examined to find optimal molar ratio of MTBE : H2O2 on extent to degradation of MTBE in groundwater at pH 7 with 10% waste ZVI. As the results, the ratio of optimization of MTBE to hydrogen peroxide for MFO was determined to be 1:300[mM]. The column experiment was conducted to know applicability of MFO-PRBs for MTBE remediation in groundwater. As the results of column test, MTBE was removed 87% of the initial concentration during 120days of operational period. Interestingly, MTBE was degraded not only within waste ZVI column but also within sand column. It means the aquifer may affect continuously the MTBE contaminated groundwater after throughout the waste ZVI barrier. The residual products showed acetone, TBF (Tert-butyl formate) and TBA (Tert-butyl acetate) during this test. The results of the present study showed that the recycled materials can be effectively used for not only a source of catalytic ferrous iron but also a reactive material of the MFO-PRBs to remove MTBE in groundwater. Key word : Waste Zero-Valent Iron, MTBE, Modified Fenton Oxidation, PRBs, MFO-PRBs

Fenton's oxidation of MTBE with zero-valent iron

Methyl tert-butyl ether (MTBE) has become a contaminant of increasing concern in the U.S. Traditional remediation technologies are successful in removing MTBE from contaminated water, but usually transfer the contaminant from the aqueous to another phase. Fenton's oxidation of MTBE provides a promising alternative to traditional remediation techniques in that it may mineralize the contaminant rather than just phase transfer. This bench-scale study investigated the feasibility of Fenton's oxidation of MTBE using zero-valent iron as the source of catalytic ferrous iron. The oxidation reactions were able to degrade over 99% of the MTBE within 10 min, and showed significant generation, and subsequent degradation, of the MTBE oxidation byproduct acetone. Second-order rate constants for MTBE degradation were 1.9×10 8 M −1 s −1 at pH 7.0 and 4.4×10 8 M −1 s −1 at pH 4.0. The total organic carbon was reduced by over 86% when a H 2O 2:MTBE ratio of 220:1 or greater was used.

Differential antigenecity of long-term in vitro cultured eutopic human endometrial stromal cells: a possible evidence of genetic disorders in endometriotic patients

Ovarian endometriosis is a recognized risk for infertility and epithelial ovarian cancer, presumably due to iron overload resulting from repeated hemorrhage. To find a clue for early detection and prevention of ovarian endometriosis-associated cancer, it is mandatory to evaluate catalytic (labile) ferrous iron (catalytic Fe(II)) and to study iron manipulation in ovarian endometriotic lesions. By the use of tissues from women of ovarian endometriosis as well as endometrial tissue from women with and without endometriosis, we for the first time performed histological analysis and cellular detection of catalytic Fe(II) with a specific fluorescent probe (HMRhoNox-M), and further evaluated iron transport proteins in the human specimens and in co-culture experiments using immortalized human eutopic/ectopic endometrial stromal cells (ESCs) in the presence or absence of epithelial cells (EpCs). The amounts of catalytic Fe(II) were higher in ectopic endometrial stromal cells (ecESCs) than in normal eutopic endometrial stromal cells (n-euESCs) both in the tissues and in the corresponding immortalized ESCs. ecESCs exhibited higher transferrin receptor 1 expression both in vivo and in vitro and lower ferroportin expression in vivo than n-euESCs, leading to sustained iron uptake. In co-culture experiments of ESCs with iron-loaded EpCs, ecESCs received catalytic ferrous iron from EpCs, but n-euESCs did not. These data suggest that ecESC play a protective role for cancer-target epithelial cells by collecting excess iron, and that these characteristics are retained in the immortalized ecESCs.

Calcium in lipid peroxidation: Does calcium interact with superoxide?

Using a pulse radiolysis approach to generate and observe superoxide anions (O − 2 ̇ ) in the absence and presence of calcium, we have attempted to verify the recent hypothesis of Babizhayev ( Arch. Biochem. Biophys. 266, 446–451, 1988) of a Ca 2+O − 2 ̇ interaction during lipid peroxidation. We could not observe rapid scavenging of O − 2 ̇ or complex formation with Ca 2+ to account for an inhibitory effect of this cation on lipid peroxidation. Neither could we agree that the stimulatory effect is due to liberation of catalytic ferrous iron from weak complexes by Ca 2+. Drawing on reports in the literature, we propose an alternate explanation for the apparent stimulation of lipid peroxidation by low Ca 2+ concentrations. In our view, this is not a direct effect, but reflects independently initiated processes of lipid peroxidation and Ca 2+ translocation, which interact subsequently in a synergistic manner. The reported inhibition at high Ca 2+ concentrations is considered an artifact as it was observed at levels far in excess of those relevant to animal systems (but not necessarily in some plant compartments).