Article

A comparison of the relative antioxidant potency of L-ergothioneine and idebenone

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Abstract

L-ergothioneine (EGT) is a stable antioxidant found in food plants as well as in animal tissue undergoing relatively high levels of oxidative stress. Idebenone is a stable analog of the antioxidant coenzyme Q(10). All are potent antioxidants found in skincare products, but their relative potencies are not well described. To establish the physiological relevance of EGT by examining transcription of the EGT transporter gene OCTN-1 and production of the receptor protein in skin fibroblasts. In addition, to compare the inhibition of lipid peroxide formation by coenzyme Q(10) and EGT. Furthermore, to compare the peroxide-scavenging abilities of EGT and idebenone in both simple solution and in cell cultures exposed to ultraviolet A (UVA). OCTN-1 expression and production in cultured fibroblasts were measured through real-time reverse transcription-PCR and Western blotting, respectively. Alloxan-induced lipid peroxidation in liposomes was used to evaluate the inhibition of lipid peroxide formation. The abilities of EGT and idebenone to directly scavenge hydroxyl radicals produced by H(2)O(2 )were determined. Finally, we irradiated fibroblasts with UVA340 radiation and compared antioxidant capabilities to scavenge free radicals. We found that OCTN-1 is expressed and readily detectable in cultured human fibroblasts. EGT was more efficient in inhibiting lipid peroxide formation than coenzyme Q(10) or idebenone. Samples treated with EGT had significantly less peroxide than those treated with idebenone 120 min after adding the antioxidants to H(2)O(2). EGT acted significantly quicker and more efficiently in capturing reactive oxygen species (ROS) after UVA340 irradiation. EGT is a natural skin antioxidant, as evidenced by the presence of the EGT transporter in fibroblasts. EGT is a more powerful antioxidant than either coenzyme Q(10) or idebenone due to its relatively greater efficiency in directly scavenging free radicals and in protecting cells from UV-induced ROS.

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... Numerous other in vitro assays and data have also demonstrated the potential antioxidant properties of EGT. Alloxan-induced lipid peroxidation of phosphatidylcholine liposomes was inhibited by 67% and 100% with the addition of only 20 μM and 100 μM EGT, respectively, which was double the protective effect of coenzyme Q 10 [76]. Park et al. [77] investigated the free radical scavenging capacity of rhizome extracts using the radical generators DPPH and ABTS, and found that high radical scavenging values were correlated with the EGT content of the extracts. ...
... The presence of OCTN1 has been reported in primary keratinocytes and fibroblast cultures and also by immunohistochemical analysis of neonatal skin sections [76,83]. Although the functional role of EGT in the skin is not known, its presence may suggest a role as a physiological protectant against ultraviolet (UV)-induced ROS generation and damage. ...
... Much of the skin damage from UV radiation is mediated through the generation of ROS [86]. Several studies have shown the cytoprotective effects of EGT against UV-induced damage and cell death [76,[87][88][89]]. Keratinocytes exposed to UV radiation showed a considerable decrease in cell viability along with a fivefold increase in caspase-9 activity, but addition of EGT was able to significantly increase cell viability and decrease caspase-9 activity [83,87]. ...
Article
Since its discovery, the unique properties of the naturally occurring amino acid, L-ergothioneine (EGT; 2-mercaptohistidine trimethylbetaine), have intrigued researchers for more than a century. This widely distributed thione is only known to be synthesized by non-yeast fungi, mycobacteria and cyanobacteria but accumulates in higher organisms at up to millimolar levels via an organic cation transporter (OCTN1). The physiological role of EGT has yet to be established. Numerous in vitro assays have demonstrated the antioxidant and cytoprotective capabilities of EGT against a wide range of cellular stressors, but an antioxidant role has yet to be fully verified in vivo. Nevertheless the accumulation, tissue distribution and scavenging properties, all highlight the potential for EGT to function as a physiological antioxidant. This article reviews our current state of knowledge. This article is part of a Special Issue entitled: Antioxidants and Antioxidant Treatment in Disease.
... Previously, we have reported that the free superoxide and singlet oxygen scavenging abilities of L-ergothioneine suppress the UV-induced TNF-α and metalloproteinase-1 expression in human dermal fibroblasts [14]. We have also documented the superior ability of 10 μM EGT compared to an equimolar solution of the water-soluble coenzyme Q10 analog idebenone to scavenge peroxides induced in NHDF by ssUV irradiation [25]. Consistent with these findings, we detected the presence of OCTN1 transcripts and protein in normal human dermal fibroblasts grown in culture [25]. ...
... We have also documented the superior ability of 10 μM EGT compared to an equimolar solution of the water-soluble coenzyme Q10 analog idebenone to scavenge peroxides induced in NHDF by ssUV irradiation [25]. Consistent with these findings, we detected the presence of OCTN1 transcripts and protein in normal human dermal fibroblasts grown in culture [25]. Prior limited evidence indicates that epidermal keratinocytes may also be capable of utilizing the antioxidant properties of EGT. ...
Article
The cellular defense system against harmful levels of reactive oxygen species consists of antioxidant enzymatic activities and small nonenzymatic molecules. L-ergothioneine has long been recognized as a potent and stable low-molecular-weight antioxidant that humans consume with diet and that accumulates in cells normally subjected to high levels of oxidative stress. As L-ergothioneine is plasma membrane-impermeative, its protective function is restricted to cells that express the L-ergothioneine-specific receptor/transporter OCTN1. Here we report for the first time that both as resident skin cells and in culture, epidermal keratinocytes synthesize OCTN1, which enables them to internalize and accumulate L-ergothioneine. This accumulation confers upon the cells an increased antioxidant potential. Consequently, it reduces the levels of reactive oxygen species and DNA, protein, and lipid damage in keratinocytes subjected to solar-simulating UV oxidative stress. Our results suggest that L-ergothioneine not only prevents oxidative damage but also may enable DNA repair in the UV-irradiated cells. The diminished oxidative damage to cellular constituents limits the apoptotic response and results in increased cell viability. The cells' ability to take up, accumulate, and utilize the potent antioxidant L-ergothioneine positions this naturally occurring amino acid and its receptor/transporter as an integral part of the antioxidative defense system of the skin.
... Stoffels et al [5] demonstrated that 1 O 2 significantly favored ESH over GSH more than 50fold for the initial reaction. Dong et al [6] indicated that addition of only 20 and 100 μM ESH could effectively inhibit alloxan-induced lipid peroxidation of phosphatidylcholine liposomes by 67% and 100%, which was more than twice the protective effect of coenzyme Q10. Pahila et al [7] demonstrated that the 1,1 diphenyl-2-picryl-hydrazyl (DPPH) radical scavenging half maximal effective concentration (IC50) of ESH could compete with ascorbic acid, and ESH revealed a stronger scavenging capacity for oxygen radicals than ascorbic acid and GSH. ...
... These results confirmed that ESH mainly contributed to the antioxidative activity of F. velutipes extract. As shown in Figure 1, chemically, ESH is an unusual thiolhistidine betaine with a sulfur atom linked to position 2 of imidazole ring [3,6]. It has distinctive features that are markedly different from ordinary thiols like GSH, for example, under physiological pH, ESH does not oxidize automatically as rapidly as GSH, and it does not promote the production of hydroxyl radicals by H 2 O 2 and Fe 2+ ions [21]. ...
Article
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Objective: The aim of this work was to assess the effect of Ergothioneine-enriched mushroom extract on oxidative stability, volatile compounds, and sensory quality of emulsified sausage. Methods: The ESH (Ergothioneine) content was determined by HPLC (High Performance Liquid Chromatography). The antioxidant activity of Flammulina velutipes (F. velutipes) extract was determined through radical-scavenging activity of 1,1 diphenyl-2-picryl-hydrazyl (DPPH), 2,2-azinobis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and hydroxyl radicals. Four different groups of emulsified sausage were manufactured: control, no antioxidants; BHA, 0.01% butylated hydroxyanisole; EEME, 0.8% ergothioneine-enriched mushroom (Flammulina velutipes) extract; AE, 0.012% authentic ergothioneine, after storage for 14 days (at 4 °C), the quality of sausage including oxidative stability (2-thiobarbituric acid reactive substances and protein carbonyls content), volatile compounds and sensory quality were studied. Results: It was demonstrated that adding ESH-enriched F. velutipes extract to sausage could effectively prevent lipid and protein oxidation, and its efficacy was equivalent with 0.01% BHA. During meat processing, the ESH mainly contributed to the antioxidative activity of F. velutipes extract. The flavor and sensory attributes of emulsified sausage were improved through adding ESH-enriched F. velutipes extract. Conclusion: Accordingly, the extract of F. velutipes contained high-level of ESH and could be a good antioxidant candidate for processed meat production.
... CoQ10, primarily found in the mitochondria, is known to play a key role in mitochondrial bioenergetics, and an antioxidant role, e.g. in prevention of DNA damage and lipid peroxidation has also been proposed ( Gruber et al., 2008;Schmelzer and Doring, 2012). Studies revealed that addition of 20 μM and 100 μM EGT inhibited alloxaninduced lipid peroxidation of phosphatidylcholine liposomes by 67% and 100%, respectively, double the protective effect of CoQ10 ( Dong et al., 2007). These antioxidant functions suggest that EGT may act by protecting vulnerable mitochondrial constituents from damage by ROS generated by the electron transport chain. ...
... This is supported by the fact that H 2 O 2 treatment of HeLa cells in which OCTN1 had been silenced provoked increased mtDNA damage (233). Furthermore, addition of EGT potentiated the protective effect of CoQ10, an antioxidant primarily found in mitochondria, in a study that assessed alloxan-induced lipid peroxidation of phosphatidylcholine liposomes (76). EGT also protects against metal-induced oxidative damage in accordance with its ability to react with bivalent metal cations such as Cu 2 + and Fe 2 + forming redox-inactive complexes (107). ...
Article
Both vitamin E (VE) consumption and blood VE status have been negatively associated with the incidence of degenerative diseases and some cancers. However, the response to VE supplementation is very variable among individuals. This could be due to interindividual variability in VE bioavailability, due, at least partly, to genetic variations in genes involved in VE metabolism. Thus, the main objective was to identify single nucleotide polymorphisms (SNPs) that may be involved in the interindividual variability in α-tocopherol (TOL) bioavailability.
... Consistently, topical use of creatine has been shown to decrease UV-induced damage in vitro and in vivo [80], and postulates it use to fight photoaging [81]. -Idebenone is a synthetic analog of coenzyme Q10 [82,83]. A clinical study using a compound based on idebenone has suggested its efficacy in preventing photoaging [84], but other studies have suggested otherwise [85,86]. ...
Article
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This review provides an overview of important concepts and trends in photoprotection. From the use of protective clothing to latest-generation oral photoprotectives, this article covers these topics from two points of view: 1) the physical blockade (absorption and/or reflection) of UV photons by topical sunscreens; 2) topical compounds with antioxidant properties that thereby protect of the consequences of UV-mediated photooxidation. The last section is devoted to the development of strong antioxidant oral compounds and discusses their possibilities as adjuvants in skin protection and repair and regeneration.
... This is supported by the fact that H 2 O 2 treatment of HeLa cells in which OCTN1 had been silenced provoked increased mtDNA damage (233). Furthermore, addition of EGT potentiated the protective effect of CoQ10, an antioxidant primarily found in mitochondria, in a study that assessed alloxan-induced lipid peroxidation of phosphatidylcholine liposomes (76). EGT also protects against metal-induced oxidative damage in accordance with its ability to react with bivalent metal cations such as Cu 2 + and Fe 2 + forming redox-inactive complexes (107). ...
Article
Significance: Mitochondrial function and specifically its implication in cellular redox/oxidative balance is fundamental in controlling the life and death of cells, and has been implicated in a wide range of human pathologies. In this context, mitochondrial therapeutics particularly involving mitochondria-targeted antioxidants have attracted increasing interest as potentially effective therapies for several human diseases. Recent advances: Over the last 10 years, great progress has been made in the development and functional testing of molecules that specifically target mitochondria, and there has been special focus on compounds with antioxidant properties. In this review, we will discuss several such strategies, including molecules conjugated with lipophilic cations (eg. TPP+) or rhodamine, conjugates of plant alkaloids, amino-acid- and peptide-based compounds and liposomes. Critical issue: This area has several major challenges that need to be confronted. Apart from antioxidants and other redox active molecules, current research aims to develop compounds capable of modulating other mitochondria-controlled processes, such as apoptosis and autophagy. Future directions: Multiple chemically different molecular strategies have been developed as delivery tools that offer broad opportunities for mitochondrial manipulation. Additional studies, and particularly in vivo approaches under physiologically relevant conditions, are necessary to confirm the clinical usefulness of these molecules.
... This is supported by the fact that H 2 O 2 treatment of HeLa cells in which OCTN1 had been silenced provoked increased mtDNA damage (233). Furthermore, addition of EGT potentiated the protective effect of CoQ10, an antioxidant primarily found in mitochondria, in a study that assessed alloxan-induced lipid peroxidation of phosphatidylcholine liposomes (76). EGT also protects against metal-induced oxidative damage in accordance with its ability to react with bivalent metal cations such as Cu 2 + and Fe 2 + forming redox-inactive complexes (107). ...
Article
Significance: Cystic fibrosis (CF) is the most common lethal genetic disorder in the Caucasian people. It is due to the mutation of cystic fibrosis transmembrane conductance regulator (CFTR) gene located on the long arm of the chromosome 7, which encodes for CFTR protein. The latter, an adenosine triphosphate binding cassette, is a transmembrane chloride channel that is also involved in glutathione transport. As glutathione/glutathione disulfide constitutes the most important pool of cellular redox systems, CFTR defects could thus disrupt the intracellular redox balance. Resulting multisystemic diseases are essentially characterized by a chronic respiratory failure, a pancreatic insufficiency, an essential fatty acid deficiency (EFAD), and inadequate levels of antioxidant vitamins. Recent advances: The pathophysiology of CF is complex; however, several mechanisms are proposed, including oxidative stress (OxS) whose implication is recognized and has been clearly demonstrated in CF airways. Critical issues: Little is known about OxS intrinsic triggers and its own involvement in intestinal lipid disorders. Despite the regular administration of pancreatic supplements, high-fat high-calorie diets, and antioxidant fat-soluble vitamins, there is a persistence of steatorrhea, EFAD, and harmful OxS. Intriguingly, several trials with elevated doses of antioxidant vitamins have not yielded significant improvements. Future directions: The main sources and self-maintenance of OxS in CF should be clarified to improve treatment of patients. Therefore, this review will discuss the potential sources and study the mechanisms of OxS in the intestine, known to develop various complications, and its involvement in intestinal lipid disorders in CF patients.
... It has been demonstrated that EGT is a more powerful antioxidant than either coenzyme Q(10) or idebenone due to its relatively greater efficiency in directly scavenging free radicals and in protecting cells from UV-induced ROS [25]. ...
Article
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Many changes related to aging at the cellular level may be due to the physiological condition of mitochondria. One of the most common types of damage of mtDNA is the so-called “common deletion” referring to a deletion of 4977 base pairs. In the skin cells this phenomenon probably is caused by oxidative damage of mtDNA induced by UV. The present study was aimed at evaluating the effect of the antioxidant l-ergothioneine on UV-induced damage in skin cells. The effect of l-ergothioneine on the reduced glutathione level was studied. The presence of the “common deletion” in human fibroblasts irradiated with UVA and treated with l-ergothioneine was evaluated by a polymerase chain reaction. We have demonstrated that l-ergothioneine enhanced the level of reduced glutathione and protected cells from the induction of a photoaging-associated mtDNA “common deletion”. In view of our results, l-ergothioneine could be an effective skin care and anti-photoaging ingredient.
... Although numerous reports detail the bioavailability of L-ergothioneine concentrations in various human tissues both in vivo, including in blood and glomerular filtration rate, [29][30][31][32] and with evaluation of protection from oxidative stress in vitro, including that in dermal fibroblasts, erythrocytes, endothelial cells, and keratinocytes, 16,20,[33][34][35] limited data are available to support ergothioneine safety. Abbreviated pharmacological studies from Tainter 36 failed to show any deleterious effects of ergothioneine at physiologically relevant concentrations. ...
Article
l-(+) Ergothioneine is a naturally occurring thiol amino acid with antioxidant properties and potential benefits as a dietary supplement. Despite its century-old identification and wide distribution in human food, little is known of its mechanism of action and safety. The nature-identical biomimetic of l-(+) ergothioneine, produced by Mironova Labs and supplied as Mironova (EGT+), has been investigated in the present studies for its mutagenic and toxicologic potential. In a plate incorporation and preincubation assay with Salmonella typhimurium strains TA98, 100, 1,535, and 1,537 and Escherichia coli WP2uvrA strain, at dose concentrations of 1.58, 5, 15.8, 50, 158, 500, 1,580, and 5,000 μg/plate with and without metabolic activation, no cytotoxicity or mutagenicity was observed. Following a preliminary 28-day study, a repeated dose 90-day gavage study at dose levels of 0, 400, 800, and 1,600 mg/kg body weight (bw)/d in Sprague Dawley rats, in which dose-proportional systemic absorption was confirmed by plasma analysis, no adverse clinical, body weight/gain, food consumption and efficiency, clinical pathology, or histopathological changes associated with the administration of the nature-identical ergothioneine were observed. In conclusion, EGT+ administered over 90 days was well tolerated with a no adverse effect level at 1,600 mg/kg bw/d, the highest dose tested for male and female rats. In addition, the nature-identical test substance, EGT+ was not mutagenic in a bacterial reverse mutation assay at plate concentrations of up to 5,000 μg/mL in the presence or absence of metabolic activation.
... Epidermal keratinocytes synthesize OCTN1, which enables ergothioneine to accumulate, and thereby confers increased antioxidant potential to skin cells. Ergothioneine scavenges reactive oxygen species (ROS) during oxidative stress in keratinocytes, increases cell viability while reducing apoptosis, as demonstrated in UVA340-irradiated cultured cells (Dong et al., 2007 ). Due to the presence of the ergothioneine transporter in fibroblasts (Grigat et al., 2007), with the resultant inhibition of lipid peroxide formation, the compound serves to provide a supportive role in skin cells as a natural skin antioxidant. ...
Article
The dietary antioxidant L-(+)-ergothioneine was tested for its potential mutagenic activity using the bacterial reverse mutation assay. The experiments were carried out using histidine-requiring auxotrophic strains of Salmonella typhimurium (Salmonella typhimurium TA98, TA100, TA1535 and TA1537), and the tryptophan-requiring auxotrophic strain of Escherichia coli (Escherichia coli WP2 uvrA) in the presence and absence of a post-mitochondrial supernatant (S9) prepared from livers of phenobarbital/β-naphthoflavone-induced rats. The revertant colony numbers of vehicle control plates with and without S9 Mix were within the corresponding historical control data ranges. The reference mutagen treatments (positive controls) showed the expected, biologically relevant increases in induced revertant colonies in all experimental phases in all tester strains. No biologically relevant increases were observed in revertant colony numbers of any of the five test strains following treatment with L-(+)-ergothioneine at any concentration level, either in the presence or absence of metabolic activation (S9 Mix) in the performed experiments. On the basis of the data reported, it can be concluded that L-(+)-ergothioneine did not induce gene mutations by base pair changes or frameshifts in the genome of the strains used. Thus L-(+)-ergothioneine has no mutagenic activity on the applied bacteria tester strains under the test conditions used in this study. Research is continuing to define the role of L-(+)-ergothioneine in disease pathophysiology. Further studies on its safety are suggested.
... Fourthly, ERG absorbs light in the ultraviolet range [123]. This physical property may account, at least in part, for its observed cytoprotective effects against UV-induced damage and cell death [34,35,41,124]. In keratinocytes exposed to ultraviolet-radiation, ERG was shown to decrease caspase-9 activity significantly, and concomitantly to increase cell survival [45,124]. However, much skin damage is mediated through the generation of reactive oxygen species, so the extent to which these results can also be explained by direct superoxide scavenging is unclear [125]. ...
Article
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Ergothioneine (ERG), is a water-soluble amino acid that is derived entirely from dietary sources. It has received much attention as a therapeutic agent due to its anti-oxidant properties, and there are claims of preferential accumulation within high oxidative stress organs. Pre-eclampsia, a condition accompanied by increased oxidative stress, is one of the leading causes of maternal morbidity and mortality. Despite intense research efforts, its aetiologies remain somewhat unclear and there are still no effective treatment options. Clinical trials of the anti-oxidants vitamin C and vitamin E have proven largely ineffective with little improvement in clinical outcome or even a negative response. This could be explained in part by their inability to permeate the plasma and mitochondrial membranes and scavenge mitochondria-derived superoxide species, and for the former by the fact that it is actually a pro-oxidant in the presence of unliganded iron. ERG accumulates within tissues through the action of a specific organic cation transporter, SLC22A4 (previously referred to as OCTN1), which is possibly also expressed in mammalian mitochondria. Mitochondrial dysfunction has been implicated in a variety of vascular diseases including pre-eclampsia. This review discusses the use of ERG as a possibly mitochondrial-targeted anti-oxidant, focusing on its physical properties, potential mechanisms of action, safety profile and administration in relation to pregnancies complicated by pre-eclampsia.
... IDB is a novel analog of coenzyme Q10 which is widely used for treatment of cardiovascular diseases (1,2). IDB can effectively inhibit lipid peroxidation in brain tissues, mitochondrial membranes and nerve cells, thus protecting cells and mitochondria from oxidative damage. ...
Article
In recent years, stem cell research has continued to benefit from its crossover with chemistry, particularly the investigation of small molecular drugs modulating specific targets to regulate stem cell fate. Idebenone (IDB) is a yellow crystalline powder that is used in the treatment of chronic cerebrovascular diseases. The objective of the present study was to examine whether IDB had an influence on bone marrow‑derived mesenchymal stem cells (BMSCs) extracted from the bone marrow of Sprague‑Dawley rats. The effects of IDB on cell proliferation, cell cloning and migration were investigated. Cell cycle, apoptosis, DAPI nuclear staining and senescence‑associated β‑galactosidase (SA‑β‑gal) staining were also examined. The results revealed that IDB at suitable concentrations enhanced cell cloning capacity, promoted the proliferation of BMSCs, delayed cellular senescence, and inhibited cell apoptosis and migration. Western blot analysis indicated that IDB increased the expression of B‑cell lymphoma 2 (Bcl‑2), signal transducer and activator of transcription‑3, Nanog, octamer‑binding transcription factor 4, E‑cadherin, proliferating cell nuclear antigen, cyclinD1 and cyclinD3, and decreased the expression of Bcl‑2‑associated X protein, cleaved caspase‑3, N‑cadherin, vimentin and α‑smooth muscle actin. In conclusion, these experiments confirmed that IDB in low doses had no toxic effect and may exert protective effects on BMSCs.
... The findings from the ex vivo studies were further supported by clinical findings of immediate and sustained hydration measurements up to 6 h and self-perceived immediate moisturizing and antifatigue effects, brighter skin, wrinkle reduction, and desire to continue using the product. Several previous studies [10][11][12][13][14][15][16][17][18] have demonstrated the actions of the individual ingredients included in the formulation objective of current investigation. Results from our study provide further evidence on this synergistic combination of ingredients. ...
Article
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Background With age, decreasing dermal levels of proteoglycans, collagen, and elastin lead to the appearance of aged skin. Oxidation, largely driven by environmental factors, plays a central role. Aim The aim of this study was to assess the antiaging efficacy of a topical serum containing L-Ascorbic acid, soluble proteoglycans, low molecular weight hyaluronic acid, and a tripeptide in ex vivo and in vivo clinical studies. Methods Photoaging and photo-oxidative damage were induced in human skin explants by artificial solar radiation. Markers of oxidative stress – reactive oxygen species (ROS), total glutathione (GSH), and cyclobutane pyrimidine dimers (CPDs) – were measured in serum-treated explants and untreated controls. Chronological aging was simulated using hydrocortisone. In both ex vivo studies, collagen, elastin, and proteoglycans were determined as measures of dermal matrix degradation. In women aged 21–67 years, hydration was measured up to 24 hours after a single application of serum, using Corneometer and hygrometer. Subjects’ perceptions of efficacy and acceptability were assessed via questionnaire after once-daily serum application for 4 weeks. Studies were performed under the supervision of a dermatologist. Results In the photoaging study, irradiation induced changes in ROS, CPD, GSH, collagen, and elastin levels; these changes were reversed by topical serum application. The serum also protected against hydrocortisone-induced reduction in collagen, elastin, and proteoglycan levels, which were significantly higher in the serum-treated group vs untreated hydrocortisone-control explants. In clinical studies, serum application significantly increased skin moisture for 6 hours. Healthy volunteers perceived the product as efficient in making the skin brighter, more hydrated, and decreasing wrinkles and wished to continue using it. The serum was well tolerated and noncomedogenic. Conclusion The serum protected against oxidative damage and dermal protein loss caused by photo- and chronological aging in human skin explants. In-vivo, the serum hydrated skin for 6 hours, and users perceived increased skin brightness, hydration, and fewer wrinkles.
... It has ability to suppress sunburn cell (SBC) formation in living skin. It has also been shown to be more effective than other commonly used topical antioxidants like a-tocopherol and co-enzyme Q10 (Dong et al. 2007; Puleo et al. 2004). Therefore the present study investigated the utility of developed Idebenone loaded nanostructured lipid carriers (INLC) for topical application as sunscreen formulation. ...
Article
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The objective of present study was to develop nanostructured lipid carriers (NLC) for topical delivery of antioxidant drug and evaluation of its sun protection efficacy. In the present study attempts have been made to formulate Idebenone loaded nanostructured lipid carriers (INLC) by using solvent precipitation method. Preformulation study evidenced for selection of Captex 500 P as an oil phase in which Idebenone has saturation solubility of 0.266 ± 0.032 g/ml. Compritol 888 ATO and ethanol were selected as solid lipid and solvent respectively. Surfactant and co-surfactant as Labrasol and Transcutol P have given stable formulations on the basis of HLB required for stabilization, respect to oil phase. INLC has particle size of 605 ± 4.01 nm and %EE of 82.58 ± 2.20 %. Optimized batches were subjected for crystallographic investigation, in vitro skin permeation study, drug deposition study, SPF determination and antioxidant activity. XRD, DSC studies illustrated that partial amorphization of Idebenone by molecularly dispersion within lipid blend leads for entrapment of drug. Permeation data showed that optimized INLC has flux value (Jss) of 7.87 μg cm−2 h−1. High significance (P < 0.001) of drug deposition in skin was observed between INLC and plain Idebenone gel. SPF value for INLC has 23 which represents that lipid nanocarriers have standards of blocking of 94–96 % of UVB rays. Such high skin deposition and SPF leads to more antioxidant effect of formulations. Hence lipid nanocarriers such as NLC have potential as an antioxidant and sun protection for topical drug delivery.
Article
Reactions involving thiol biochemistry seem to play a crucial role in skeletal muscle fatigue. N-acetylcysteine amide (NACA) and L-ergothioneine (ERGO) are thiol-based antioxidants available for human use that have not been evaluated for effects on muscle fatigue. To test the hypothesis that NACA and ERGO delay skeletal muscle fatigue. We exposed mouse diaphragm fiber bundles to buffer (CTRL), NACA, ERGO, or N-acetylcysteine (NAC; positive control). Treatments were performed in vitro using 10 mM for 60 min at 37 °C. After treatment, we determined the muscle force-frequency and fatigue characteristics. The force-frequency relationship was shifted to the left by ERGO and to the right by NACA compared with CTRL and NAC. Maximal tetanic force was similar among groups. The total force-time integral (FTI; N · s · cm) during the fatigue trial was decreased by NACA (420 ± 35, P < 0.05), unaffected by ERGO (657 ± 53), and increased by NAC (P < 0.05) compared with CTRL (581 ± 54). The rate of contraction (dF/dtMAX) during the fatigue trial was not affected by any of the treatments tested. NAC, but not NACA or ERGO, delayed the slowing of muscle relaxation (dF/dtMIN) during fatigue. In summary, NACA and ERGO did not delay skeletal muscle fatigue in vitro. We conclude that these antioxidants are unlikely to improve human exercise performance.
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In this review, we provide a wide, non-comprehensive overview of the latest trends in protection against the deleterious effects of the sun. The review is divided into physical blockade of UV photons, UV photon absorption and/or reflection provided by topical sunscreens and topical antioxidant compounds. The last section is devoted to the newest generation of dietary photoprotective compounds. Possible mechanisms, particularly antioxidant, are discussed.
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The ability of a hydrophilic extract prepared from edible mushroom (Flammulina velutipes) to stabilize fresh color of bigeye tuna (Thunnus obesus) meat was evaluated to compare it with certain other antioxidants. The fresh color shelf life of bigeye tuna meats, to which were added as 1, 3, or 5 mL of mushroom extract to 100 g of minced bigeye tuna meat, prolonged duration of ice storage by more than 2, 4, and 6 d, respectively, in comparison with the control tuna meat without mushroom extract. The addition of 5 mL of mushroom extract to 100 g of minced bigeye tuna meat was more effective than adding ascorbic acid sodium salt (500 ppm) or α-tocopherol (500 ppm) with regard to oxidation of lipid in the tuna meat. The color changes significantly correlated with lipid oxidation as well as metmyoglobin formation in the tuna meat. These results clearly show that the mushroom extract is a potential antioxidant, which has the ability to stabilize fresh color of tuna meat during ice storage.
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The native amino acid ergothioneine, a thiourea derivative of histidine, inhibits mushroom tyrosinase activity in a dose-dependent manner, with an IC(50) value of 1.025 mg/ml (4.47 mM). By contrast, histidine exhibited no inhibitory effect on mushroom tyrosinase activity. We characterized ergothioneine as a noncompetitive tyrosinase inhibitor using a Lineweaver-Burk plot of experimental kinetic data. The IC(50) value for ergothioneine scavenging of 2,2-diphenyl-1-picrylhydrazyl was 6.110 ± 0.305 mg/ml, much higher than the IC(50) for inhibition of tyrosinase activity which indicating ergothioneine on tyrosinase shows a weak correlation to its antioxidative activity. The results demonstrated that ergothioneine has a potent inhibition effect on tyrosinase enzyme activity, resulting from the presence of the sulfur substituted imidazole ring in ergothioneine.
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The Acute Respiratory Distress Syndrome (ARDS), the most severe form of Acute Lung Injury (ALI), is a highly-fatal, diffuse non-cardiogenic edematous lung disorder. The pathogenesis of ARDS is unknown but lung inflammation and lung oxidative stress are likely contributing factors. Since no specific pharmacologic intervention exists for ARDS, our objective was to determine the effect of treatment with ergothioneine-a safe agent with multiple anti-inflammatory and antioxidant properties on the development of lung injury and inflammation in rats insufflated with cytokines found in lung lavages of ARDS patients. Sprague-Dawley rats (3-10/group) were given 15 mg/kg or 150 mg/kg l-ergothioneine intravenously 1h before or 18 h after cytokine (IL-1 and IFNγ) insufflation. Lung injury (lavage LDH levels) and lung inflammation (lavage neutrophil numbers) were measured 24h after cytokine insufflation. Ergothioneine pre- and post-treatment generally decreased lung injury and lung inflammation in cytokine insufflated rats. Ergothioneine should be considered for additional testing as a potential therapy for treating and preventing ARDS.
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Using (1)H-nuclear magnetic resonance (NMR) spectroscopy and statistical models, we sought to identify ''biomarkers'' present in erythrocytes that would distinguish between women with normal pregnancy and those suffering from preeclampsia, and investigate possible links with previously identified plasma ''markers.'' Erythrocytes from 22 normotensive pregnant women and 15 preeclamptics were analyzed by (1)H Carr-Purcell-Meiboom-Gill (CPMG) NMR. Multivariate analysis and logistic regression were applied to differentiate between the 2 groups of patients, and used to develop a diagnostic model based on the concentrations of the constituents identified as being influential. Significantly higher concentrations of alanine (P < .001), glycine (P = .025), and ergothioneine (P = .049) were found in erythrocytes from preeclamptic patients. Discriminant analysis and regression of NMR data permitted 100% accurate diagnosis of the health status of new patients. Chemically related imidazole-based molecules, histidine and ergothioneine, are important in the classification process and the etiology of preeclampsia (PE).
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New developments in the realm of skin rejuvenation such as phytotherapy are at an astounding increasing pace in the cosmeceutical market. Yet, many of these products that are classified as cosmeceuticals are tested less vigorously and do not have to be approved by the Food and Drug Administration to establish efficacy and safety. Thus, as clinicians, we must ask the question, “Is there science-based evidence to validate the mechanism of these new treatments?” We assessed the top anti-aging creams currently on the market specifically evaluating their botanical ingredients. Some of the most common botanicals that are hot off the market are: Rosmarinus officinalis, Vitis vinifera (grape seed extract), Citronellol, Limonene, Oenothera biennis (evening primrose), Glycyrrhiza glabra (licorice extract), Aframomum angustifolium seed extract, Diosgenin (wild yam), N6 furfuryladenine (kinetin), and Ergothioneine. Through researching each of these botanical ingredients, we have concluded that randomized controlled trials are still needed in this area, but there is promise in some of these ingredients and science to validate them.
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Ultraviolet B (UVB) irradiation can cause human skin damage or skin aging and wrinkle formation through photochemical reactions. Antioxidative substances may ameliorate UV damage. In this study, the anti-photoaging activity of three antioxidants—ergothioneine, ferulic acid, and glutathione—was investigated after UVB irradiation of Hs68 human skin fibroblast cells. The cells treated with these three antioxidants appeared similar to unirradiated control cells. UVB irradiation decreased cell viability by 26% compared to that of unirradiated control cells. However, the addition of either single or combined antioxidants enhanced cell viability after UVB irradiation. These three antioxidants can inhibit the production of reactive oxygen species (ROS) induced by the UVB irradiation of the Hs68 cells. Ergothioneine showed a greater inhibitory effect on matrix metalloproteinase-1 (MMP-1) performance than the other two antioxidants. IL-1 alpha was not detected in the Hs68 cells after exposure to a radiation dose of 150 mJ/cm2. Ergothioneine showed better restoration of type 1 procollagen than either ferulic acid or glutathione. Based on these results, the addition of two antioxidants was expected to restore type Ι procollagen production. In summary, these results demonstrate that the three tested antioxidants protect the skin against UVB-induced damage. The single and combined use of ergothioneine, ferulic acid, and glutathione has the potential for development as anti-photoaging materials in cosmetic applications.
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The aim of this study was to compare the effects of L-ergothioneine (LE) and L-ascorbic acid (LAA) on the IVM of immature oocytes and embryonic development of in vitro matured oocytes in sheep. The ovaries were collected from slaughterhouse, then they were transferred (within 1-2 hs) to the laboratory at warm (30-35°C) 0.9% saline solution. Cumulus Oocyte Complexes (COCs) were obtained by incision of follicles. Oocytes covered with at least three compact layers of cumulus oophorus cell were in vitro matured in TCM 199 with 10% Foetal Calf Serum (FCS) (v/v), Follicle Stimulating Hormone (FSH), Luteinizing Hormone (LH) and Na Pyruvate with supplemented of 10 mM of LE (Group 1), 10 mM LAA (Group 2) or without any antioxidant (Group 3: Control) at 24 h. COCs with expanded cumulus layers following IVM were transferred to IVF TALP micro-drops with or without antioxidants (LE or LAA). Fresh semen from ram were separated by Percoll-Gradient and capacitated with Hepes TALP with heparine (30 IU/ml). Then, separated and capacitated spermatozoa were transferred to IVF TALP micro-drops containing matured oocytes and incubated for 20 hs. Presumptive zygotes were in vitro cultured in TCM 199 with or without antioxidants (LE or LAA) in 7-8 days. The results showed that, in LE group, the percentages of maturation, 8 -cell and morula stages were significantly higher than those of LAA and control groups (P<0.01-0.05) except for cleavage rate which had a numerically higher (P≥0.05). In conclusion, addition of L-ergothionine into IVM, IVF and IVC medium had a beneficial effect on in vitro maturation of oocytes and embryonic development, especially from cleavage to morula stages compared to LAA and Control groups.
Article
This study investigates the effect of insulin combined with idebenone on blood–brain barrier (BBB) permeability in experimental streptozotocin-induced diabetic rats as well as the underlying mechanisms. With a diabetic rat model, we show that insulin and idebenone normalize body weight and water intake and restore BBB permeability and that their combination displays a synergistic effect. The results from transmission electron microscopy show that the combination of insulin and idebenone significantly closed the tight junction (TJ) in diabetic rats. The results from Western blotting in diabetic rats show that the upregulation of TJ-associated proteins occludin, and zonula occludens (ZO)-1 caused by the combination of insulin and idebenone is more remarkable than that with either agent alone. In addition, the activations of reactive oxygen species (ROS) and advanced glycation end products (AGEs) and the expression levels of receptors for advanced glycation end-products (RAGE) and nuclear factor-κB (NF-κB) were significantly decreased after treatment with insulin and idebenone in diabetic rats. These results suggest that the combination of insulin and idebenone could decrease the BBB permeability in diabetic rats by upregulating the expression of occludin, claudin-5, and ZO-1 and that the ROS/AGE/RAGE/NF-κB signal pathway might be involved in the process. © 2014 Wiley Periodicals, Inc.
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Although berberine (BBR) shows antioxidant activity, its activity is limited. We synthesized 9-O-benzoic acid berberine derivatives, and their antioxidant activities were screened via ABTS, DPPH, HOSC and FRAP assays. The para-position was modified with halogen elements on the benzoic acid ring, which led to an enhanced antioxidant activity and the substituent on the ortho-position was found to be better than the meta-position. Compounds 8p, 8c, 8d, 8i, 8j, 8l, and especially 8p showed significantly higher antioxidant activities, which could be attributed to the electronic donating groups. All the berberine derivatives possessed proper lipophilicities. In conclusion, compound 8p is a promising antioxidant candidate with remarkable elevated antioxidant activity and moderate lipophilicity.
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L-ergothioneine is an amino acid synthetized by fungi and mycobacteria which cannot be synthesized by other species. It has been detected in plants, animals and the human body. In the last past years, it has been recognized as a good antioxidant and recently, it has also been related to other properties besides antioxidant properties. Even though few studies on the toxicity of L-ergothioneine have been carried out, evidence suggests that L-ergothioneine is not harmful to health. Considering that L-ergothioneine is increasingly been linked to positive effects on human health coupled with the fact that it seems to be safe for human consumption, this molecule may be suitable for use as an ingredient in foods. On the other hand, despite the positive effects reported for this molecule, no estimate of L-ergothioneine intake has been carried out until now. Thus, the aim of this work is to estimate the intake of L-ergothioneine through food consumption of several European Countries and the United States. Values were estimated by using the deterministic and probabilistic approach. Results shown that the populations with the highest intake of L-ergothioneine correspond to Italian population, both for children and adults.
Article
An efficient and sensitive method was established for quantitative analysis of L-Ergothioneine during fermentation analyze by reversed-phase-high performance liquid chromatography (RP-HPLC). The method was carried out on two C18 columns (4.6 × 250 mm, 5 um), and the isocratic mobile phase was 1 % methanol containing boric acid adjusted to a pH of 5.0 with a flow rate of 0.7 mL/min. An UV-VIS detector equipped with a wavelength of 257 nm was employed. The injection volume was 5 μL, with the columns temperature being 25 °C. The linearity, recovery, limit of detection (LOD) and quantification (LOQ), precision, repeatability, stability, and recovery were all tested and good results were obtained. The method was simple, rapid, accurate, and high sensitivity and could be utilized for the research and development of L-Ergothioneine in industry.
Chapter
Better understanding of molecular damage caused by solar radiation has led to the growing study of molecules with antioxidant and anti-inflammatory activity against photodamage. Oral use of these substances is considered an important coadjutant in photoprotection strategy, due to two general mechanisms: prevention and mitigation of photodamage. Coming from nutrients, such as vitamins and functional foods, or herbal extracts, and, more recently, probiotics, molecules with oral photoprotection action have antioxidant action, protecting especially from damage to DNA and protein and lipid structures, but they can also prevent or mitigate UV-induced inflammation, acting on the epidermis and dermis. An increase in the minimal erythema dose has also been shown with some associations of nutrients. Although the level of evidence varies considerably between the molecules studied and described, the most-used active constituents with proven safety and efficacy under systemic use are presented below. These molecules act most often in association, providing a synergistic effect that also allows reduction of their respective concentrations and, consequently, greater tolerability for use.
Chapter
Better understanding of molecular damage caused by solar radiation has led to the growing study of molecules with antioxidant and anti-inflammatory activity against photodamage. Oral use of these substances is considered an important coadjutant in photoprotection strategy, due to two general mechanisms: prevention and mitigation of photodamage. Coming from nutrients, such as vitamins and functional foods, or herbal extracts, and, more recently, probiotics, molecules with oral photoprotection action have antioxidant action, protecting especially from damage to DNA and protein and lipid structures, but they can also prevent or mitigate UV-induced inflammation, acting on the epidermis and dermis. An increase in the minimal erythema dose has also been shown with some associations of nutrients. Although the level of evidence varies considerably between the molecules studied and described, the most-used active constituents with proven safety and efficacy under systemic use are presented below. These molecules act most often in association, providing a synergistic effect that also allows reduction of their respective concentrations and, consequently, greater tolerability for use.
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Ergothioneine (ERG) is an unusual thio-histidine betaine amino acid that has potent antioxidant activities. It is synthesised by a variety of microbes, especially fungi (including in mushroom fruiting bodies) and actinobacteria, but is not synthesised by plants and animals who acquire it via the soil and their diet, respectively. Animals have evolved a highly selective transporter for it, known as solute carrier family 22, member 4 (SLC22A4) in humans, signifying its importance, and ERG may even have the status of a vitamin. ERG accumulates differentially in various tissues, according to their expression of SLC22A4, favouring those such as erythrocytes that may be subject to oxidative stress. Mushroom or ERG consumption seems to provide significant prevention against oxidative stress in a large variety of systems. ERG seems to have strong cytoprotective status, and its concentration is lowered in a number of chronic inflammatory diseases. It has been passed as safe by regulatory agencies, and may have value as a nutraceutical and antioxidant more generally.
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The aim of this study was to compare the effects of L-ergothioneine (LE) and L-ascorbic acid (LAA) on the IVM of immature oocytes and embryonic development of in vitro matured oocytes in sheep. The ovaries were collected from slaughterhouse, then they were transferred (within 1-2 hs) to the laboratory at warm (30-35°C) 0.9% saline solution. Cumulus Oocyte Complexes (COCs) were obtained by incision of follicles. Oocytes covered with at least three compact layers of cumulus oophorus cell were in vitro matured in TCM 199 with 10% Foetal Calf Serum (FCS) (v/v), Follicle Stimulating Hormone (FSH), Luteinizing Hormone (LH) and Na Pyruvate with supplemented of 10 mM of LE (Group 1), 10 mM LAA (Group 2) or without any antioxidant (Group 3: Control) at 24 h. COCs with expanded cumulus layers following IVM were transferred to IVF TALP micro-drops with or without antioxidants (LE or LAA). Fresh semen from ram were separated by Percoll-Gradient and capacitated with Hepes TALP with heparine (30 IU/ml). Then, separated and capacitated spermatozoa were transferred to IVF TALP micro-drops containing matured oocytes and incubated for 20 hs. Presumptive zygotes were in vitro cultured in TCM 199 with or without antioxidants (LE or LAA) in 7-8 days. The results showed that, in LE group, the percentages of maturation, 8 -cell and morula stages were significantly higher than those of LAA and control groups (P<0.01-0.05) except for cleavage rate which had a numerically higher (P≥0.05). In conclusion, addition of L-ergothionine into IVM, IVF and IVC medium had a beneficial effect on in vitro maturation of oocytes and embryonic development, especially from cleavage to morula stages compared to LAA and Control groups.
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Ozone, the main component of photochemical smog and air pollution, can damage the skin by oxidizing stratum corneum enzymes, lipids and structural proteins. We have developed a rapid screening assay to determine free radical scavenging capacity of various active ingredients that are frequently used in personal care products. Several known antioxidants including vitamin C, vitamin E analog Trolox, walnut seed extract, lipoic acid and ergothioneine inner salt were assayed for their ability to neutralize ozone-induced oxidation of beta-phycoerythrin, a fluorescent reporter protein derived from algae. The free radical scavenging capacities of these antioxidants were quantified and compared. The results demonstrate that this assay is a valuable primary screening tool for identifying antioxidant activity of natural or synthetic substrates that can be used in personal care products to protect the uppermost layer of our skin from oxidizing damage induced by O3.
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Variants of the SLC22A4 gene are associated with susceptibility to rheumatoid arthritis and Crohn's disease. SLC22A4 codes for an integral membrane protein, OCTN1, that has been presumed to carry organic cations like tetraethylammonium across the plasma membrane. Here, we show that the key substrate of this transporter is in fact ergothioneine (ET). Human OCTN1 was expressed in 293 cells. A substrate lead, stachydrine (alias proline betaine), was identified by liquid chromatography MS difference shading, a new substrate search strategy. Analysis of transport efficiency of stachydrine-related solutes, affinity, and Na⁺ dependence indicates that the physiological substrate is ET. Efficiency of transport of ET was as high as 195 μl per min per mg of protein. By contrast, the carnitine transporter OCTN2 from rat did not transport ET at all. Because ET is transported >100 times more efficiently than tetraethylammonium and carnitine, we propose the functional name ETT (ET transporter) instead of OCTN1. ET, all of which is absorbed from food, is an intracellular antioxidant with metal ion affinity. Its particular purpose is unresolved. Cells with expression of ETT accumulate ET to high levels and avidly retain it. By contrast, cells lacking ETT do not accumulate ET, because their plasma membrane is virtually impermeable for this compound. The real-time PCR expression profile of human ETT, with strong expression in CD71⁺ cells, is consistent with a pivotal function of ET in erythrocytes. Moreover, prominent expression of ETT in monocytes and SLC22A4 polymorphism associations suggest a protective role of ET in chronic inflammatory disorders. • erythrocyte • inflammation
Chapter
This chapter presents a brief review of the accumulated knowledge concerning ergothioneine. Although several biological activities have been suggested for ergothioneine in the past few years, the full meaning of these activities is not yet clear. It is quite likely that its chief function or functions have not yet been uncovered. It is obviously not yet possible to classify ergothioneine in the realm of hiologically active compounds such as vitamins or hormones. It now appears fairly certain that ergothioneine is not synthesized by higher animals. On the other hand, experiments so far have failed to demonstrate that animals need the compound for survival under normal laboratory conditions. At the present state of knowledge, it would appear that ergothioneine cannot be classified as a vitamin in the usual sense of the term. On the other hand, recent developments have suggested that ergothioneine may have regulatory effects on enzyme activity. In addition, a possibility which has not yet been explored is the effect of ergothioneine on animals under various stress conditions. It is within the realm of possibility that the substance cannot be neatly categorized in any of the recognized classes of biologically active compounds; indeed, it is perhaps for this reason that its function or functions have defied elucidation for so long.
Article
The high performance liquid chromatographic procedure was established for the determination of ergothioneine in biological materials. The procedure is also applicable to the purification of 3H-ergothioneine. The metabolism of ergothioneine given to rats was very slow ; the ergothioneine was not disappear even after 1 week fasting. The ergothioneine administrated was mostly concentrated in the liver and the level was 10 times as high as that in blood. All ergothioneine in the blood was found in blood corpuscles 24 hr after the administration.
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Recently a number of possible functions for ergothioneine (ERT) have been suggested (1). This paper elaborates on some of these in light of overlooked or recent publications and presents additional hypotheses including: 1. Reduced ERT (ergothionol) may be an acyl carrier. 2. ERT, in conjunction with thyroid hormone and iodine, may be a cofactor in peroxidative and oxidative reactions. 3. ERT and thyroid hormone may be required for the oxidation of reduced pyridine nucleotides and the coupling of this to oxygen consumption (respiration) and ATP generation/ATPase action (heat production). 4. ERT may be required for both gene expression and repair. 5. 2-Thioimidazoles (ERT and 2-thiourocanic acid in particular) may be immunoregulatory. 6. ERT may be involved in the protection from oxidation (inactivation) of methionine and methionine containing chemoattractants, hormones, tRNA, etc. Some future research activities are suggested.
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Since its discovery at the turn of the century, attempts to define a physiological function for L-ergothioneine have been unsuccessful. This paper suggests several possible functions for this enigmatic compound or its metabolites. These include: transport of cations or carbon dioxide, catalysis of carboxylation or decarboxylation reactions, mediation of thyroid or antithyroid function, histaminic or antihistaminic action, and cholinergic or anticholinergic action.
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Calf skin acid-soluble collagen in microfibrillar form was incubated with free oxygen radicals produced by the system xanthine oxidase + hypoxanthine. This incubation liberated peptides of a size smaller than that of alpha-chains, as demonstrated by SDS-PAGE and by evaluation of the 4-hydroxyproline contained in small peptides. The amount of liberated peptides was found to increase with time. The process was inhibited by addition of superoxide dismutase to the medium but not by addition of catalase. Two flavonoids extracted from bilberries and a third one from grapes were demonstrated to protect collagen against this non-enzymatic proteolytic activity. This work confirms that collagen may be degraded during the process of inflammation and that some flavonoids are endowed with protective properties.
Article
Advanced glycation end-products (AGEs) have been reported to be accumulated in dermal skin. However, the role of AGEs in the photoaging of human skin remains unknown, and for this reason, we have examined the interaction between AGEs and ultraviolet A light (UVA) from both the chemical and biological aspects. Previously, we reported that exposing human dermal fibroblasts to UVA in the presence of AGEs that were prepared with bovine serum albumin (BSA) decreased the cell viability due to superoxide anion radical s (.O2(-)) and hydroxyl radicals (.OH) generated by AGEs under UVA irradiation, and active oxygen species are detected with ESR spin-trapping. To identify the active oxygen species in detail and to clarify the cell damaging mechanism, we performed several experiments and the following results were obtained. (1) In ESR spin-trapping, by addition of dimethyl sulfoxide and superoxide dismutase, ESR signals due to .O2(-) -derived DMPO-OOH and .OH-derived DMPO-OH adducts, respectively, were detectable. (2) UVA-irradiated AGEs elevated the lipid peroxide levels in both fibroblasts and liposomes. But the peroxidation in liposomes was inhibited by addition of deferoxamine. (3) Survival of fibroblasts exposed to UVA in the presence of AGEs was elevated by addition of deferoxamine. And finally, (4) survival of fibroblasts was found to be regulated by the level of H2O2. On the basis of these results, we propose a possible mechanism in which AGEs under UVA irradiation generate active oxygen species involving .O2(-), H2O2, and .OH, and the .OH species plays a harmful role in promoting cell damage.
Article
The organic cation transporter OCTN1 (SLC22A4) is expressed ubiquitously, with strong expression in kidney, trachea, bone marrow, and fetal liver, and it mediates transport of organic cations in a pH-dependent manner. Recent studies have identified single nucleotide polymorphisms (SNPs) of OCTN1 in the Japanese population. Two SNPs present in the exon regions, c1063t and g1531a, cause amino acid mutation, Thr306Ile (T306I) and Gly462Glu (G462E), respectively. We examined the influence of these SNPs on the intracellular localization, protein expression, and transport activity of OCTN1. Immunocytochemical analysis showed similar localizations of OCTN1 in cellular membranes of HEK293 cells transiently transfected with an expression plasmid DNA for OCTN1 or its SNP allelic variants. The Km and Vmax values for tetraethylammonium (TEA) uptake by T306I were similar to those of the wild-type even when the Vmax value was normalized for the expression level of OCTN1 protein. In contrast, G462E had almost negligible transport activity, although the protein expression level of G462E was equivalent to that of the wild-type. We conclude that the SNP that causes the single amino acid mutation T306I does not affect TEA transport activity, whereas the mutation G462E abrogates the TEA transport activity, presumably affecting the physiological function of OCTN1 and/or the pharmacological characteristics of its substrates.
Article
Ergothioneine (EGT) is a sulfur-containing amino acid, and is presumed to function as a natural antioxidant. The purpose of this study was to identify the nature of the antioxidant activity and investigate the effects of EGT on UV-induced cellular response. In chemical studies, EGT scavenged the superoxide anion radical (*O(2)(-)) and singlet oxygen ((1)O(2)). In cultured fibroblasts, EGT suppressed TNF-alpha up-regulation by UVB irradiation. In addition, in fibroblasts exposed to UV-A, EGT suppressed the expression of matrix metalloproteinase 1 (MMP-1) protein by nearly 50% and reduced MMP-1 mRNA expression. From these results, we conclude that EGT scavenges reactive oxygen species generated by both Type I and Type II photosensitization and suppresses both TNF-alpha expression and MMP-1 at their transcriptional level. EGT may reduce skin anti-aging effects after UV irradiation by the scavenging of *O(2)(-) and (1)O(2), and reducing signals for protease and inflammatory activity.
Article
Treatment of oxidative stress-related pathologies is a possible therapeutical strategy for the future. Natural product with antioxidant properties could trigger this goal. The aim of this in vitro study was to assess the antioxidant activity of the natural product ergothioneine (EGT), a compound of plant origin, which is assimilated and conserved by mammals in erythrocytes, kidney, seminal fluid and liver. We measured the antioxidant activity of EGT as its ability to antagonize the oxidation of alpha-keto-gamma-methiolbutyric acid (KMBA) by hydroxyl radical, peroxyl radicals and peroxynitrite. The results are expressed as total oxyradical scavenging capacity (TOSC) units. Glutathione (GSH), uric acid and 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (trolox), the water-soluble analog of vitamin E, were used as the reference antioxidants. EGT was the most active scavenger of free radicals as compared to classic antioxidants as GSH, uric acid and trolox. In particular, the highest antioxidant capacity exhibited by EGT vs. peroxyl radicals (5.53 +/- 1.27 units) resulted 25% higher than the value obtained with the reference antioxidant trolox (4.4 +/- 0.6 units, P < 0.01). The scavenging capacity of EGT towards hydroxyl radicals (0.34 +/- 0.09 units) was 60% higher, as compared to uric acid (0.21 +/- 0.04 units, P < 0.001), which represent the reference antioxidant vs. hydroxyl radicals. Finally, EGT showed the highest antioxidant activity also towards peroxynitrite (5.2 +/- 1.0 units), with a scavenging capacity 10% higher than that of uric acid (4.7 +/- 0.9 units, P < 0.05). This study showed that EGT has potent intrinsic anti-hydroxyl, anti-peroxyl and anti-peroxynitrite radicals antioxidant activity, as compared to classic molecules with antioxidant capacity as GSH, trolox and uric acid. This appears of interest, given the increasing use of non-vitamins cocktails for therapeutical approaches to many oxidative-induced pathologies.
Article
Idebenone is an antioxidant lower molecular weight analogue of coenzyme Q10. Previously, idebenone was shown to be a very effective antioxidant in its ability to protect against cell damage from oxidative stress in a variety of biochemical, cell biological, and in vivo methods, including its ability to suppress sunburn cell (SBC) formation in living skin. However, no clinical studies have been previously conducted to establish the efficacy of idebenone in a topical skincare formulation for the treatment of photodamaged skin. In this nonvehicle control study, 0.5% and 1.0% idebenone commercial formulations were evaluated in a clinical trial for topical safety and efficacy in photodamaged skin. Forty-one female subjects, aged 30-65, with moderate photodamaged skin were randomized to use a blind labelled (either 0.5% or 1.0% idebenone in otherwise identical lotion bases) skincare preparation twice daily for six weeks. Blinded expert grader assessments for skin roughness/dryness, fine lines/wrinkles, and global improvement in photodamage were performed at baseline, three weeks and six weeks. Electrical conductance readings for skin surface hydration and 35 mm digital photography were made at baseline after six weeks. Punch biopsies were taken from randomly selected subjects, baseline and after six weeks, and stained for certain antibodies (interleukin IL-6, interleukin IL-1b, matrixmetalloproteinase MMP-1, collagen I) using immunofluorescence microscopy. After six weeks' use of the 1.0% idebenone formula, a 26% reduction in skin roughness/dryness was observed, a 37% increase in skin hydration, a 29% reduction in fine lines/wrinkles, and a 33% improvement in overall global assessment of photodamaged skin. For the 0.5% idebenone formulation, a 23% reduction in skin roughness/dryness was observed, a 37% increase in skin hydration, a 27% reduction in fine lines/wrinkles, and a 30% improvement in overall global assessment of photodamaged skin. The immunofluorescence staining revealed a decrease in IL-1b, IL-6, and MMP-1 and an increase in collagen I for both concentrations.
Article
Topical applications of skin care products containing antioxidants have become increasingly popular. Numerous studies have elucidated the biological effects of these substances. General antiaging effects, anti-inflammatory properties, photoprotective properties, and prevention of ultraviolet (UV) immunosuppression have been documented. However, a standardized method to characterize and compare the properties and oxidative stress protection capacity of antioxidants was lacking. A multistep in vitro process utilizing a variety of biochemical and cell biological methods combined with in vivo studies was designed to compare the oxidative stress protective capacity of commonly used antioxidants. Data were presented for L-ascorbic acid, dl-alpha-tocopherol, kinetin, dl-alpha lipoic acid, ubiquinone, and idebenone. Methods included using UV-induced radical trapping/scavenging capacity measured by photochemiluminescence, pro-oxidative systems (LDL-CuSO(4), microsome-NADPH/ADP/Fe(3+)) with measurement of primary and secondary oxidation products, UVB irradiation of human keratinocytes, and in vivo evaluation, using the human sunburn cell (SBC) assay. Correlation and trends between in vitro and in vivo results were established, and the standardized test protocol was used to quantify oxidative stress protection capacity of antioxidants. Summarizing and totaling the data equally weighted for each oxidative stress study, the overall oxidative protection capacity scores of 95, 80, 68, 55, 52, and 41 were obtained for idebenone, dl-alpha tocopherol, kinetin, ubiquinone, L-ascorbic acid, and dl-alpha lipoic acid, respectively. The higher the score, the more effective the overall oxidative stress protection capacity of the antioxidant became. This multistep protocol may serve as a standard in investigating and comparing new putative antioxidants for topical use as well as a valuable tool to assess the anti-inflammatory properties, photoprotective properties, and prevention of UV immunosuppression of topical antioxidants.