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Using a new red blood cell (RBC) metabolite extraction protocol, we performed a metabolomic analysis on RBCs in rheumatoid arthritis (RA) patients treated or not with methotrexate (MTX), with the two following objectives: to compare the RBC metabolic profiles of MTX-naïve RA patients and healthy controls (HC), and to investigate whether RBC profiles before and after MTX treatment in RA differed between responders and non-responders. Plasma analysis was performed in parallel. Metabolites were extracted and identified in RBCs and plasma by liquid chromatography-mass spectrometry. We compared the metabolomic fingerprints of 31 DMARD-naïve RA patients and 39 HCs. We also compared the RBC and plasma metabolomes of 25 RA patients who responded or not to MTX therapy before (M0) and after a 3-month treatment period (M3). Significance was determined by Storey's false discovery rate (FDR) q-values to correct for multiple testing. RA patients and HCs differed in the metabolomic signature of RBCs. The signature mainly contained amino acids (AA). Eleven metabolites, including 4 metabolites belonging to the carbohydrate subclass and 2 amino acids (creatine and valine) showed accumulation in RBCs from RA patients. Conversely, citrulline (fold change = 0.83; q = 0.025), histidine (fold change = 0.86; q = 0.014) and ergothioneine (EGT) (fold change = 0.66; q = 0.024), were lower in RBC of RA patients. Five plasma metabolites, including succinic acid and hydroxyproline, were higher in RA patients, and 7 metabolites, including DHEA sulfate, alanine, threonine and ornithine, were lower. Among RA patients undergoing MTX treatment pre-treatment (M0), EGT values were significantly lower in non-responders. In conclusion, low RBC levels of EGT, a food-derived AA barely detectable in plasma, characterize DMARD naïve RA patients and lack of response to MTX treatment. [ABSTRACT FROM AUTHOR]

Healthy aging has emerged as a crucial issue with the increase in the geriatric population worldwide. Food-derived sulfur-containing amino acid ergothioneine (ERGO) is a potential dietary supplement, which exhibits various beneficial effects in experimental animals although the preventive effects of ERGO on aging and/or age-related impairments such as frailty and cognitive impairment are unclear. We investigated the effects of daily oral supplementation of ERGO dissolved in drinking water on lifespan, frailty, and cognitive impairment in male mice from 7 weeks of age to the end of their lives. Ingestion of 4 ~ 5 mg/kg/day of ERGO remarkably extended the lifespan of male mice. The longevity effect of ERGO was further supported by increase in life and non-frailty spans of Caenorhabditis elegans in the presence of ERGO. Compared with the control group, the ERGO group showed significantly lower age-related declines in weight, fat mass, and average and maximum movement velocities at 88 weeks of age. This was compatible with dramatical suppression by ERGO of the age-related increments in plasma biomarkers (BMs) such as the chemokine ligand 9, creatinine, symmetric dimethylarginine, urea, asymmetric dimethylarginine, quinolinic acid, and kynurenine. The oral intake of ERGO also rescued age-related impairments in learning and memory ability, which might be associated with suppression of the age-related decline in hippocampal neurogenesis and TDP43 protein aggregation and promotion of microglial shift to the M2 phenotype by ERGO ingestion. Ingestion of ERGO may promote longevity and healthy aging in male mice, possibly through multiple biological mechanisms. [ABSTRACT FROM AUTHOR]

Sixteen strains of basidiomycetous yeasts were evaluated for their capability to produce ergothioneine (EGT), an amino acid derivative with strong antioxidant activity. The cells were cultured in either two synthetic media or yeast mold (YM) medium for 72 h, after which cytosolic constituents were extracted from the cells with hot water. After analyzing the extracts via liquid chromatography-mass spectrometry (LC-MS), we found that all strains produced varying amounts of EGT. The EGT-producing strains, including Ustilago siamensis, Anthracocystis floculossa, Tridiomyces crassus, Ustilago shanxiensis, and Moesziomyces antarcticus, were subjected to flask cultivation in YM medium. U. siamensis CBS9960 produced the highest amount of EGT at 49.5 ± 7.0 mg/L after 120 h, followed by T. crassus at 30.9 ± 1.8 mg/L. U. siamensis was also cultured in a jar fermenter and produced slightly higher amounts of EGT than under flask cultivation. The effects of culture conditions, particularly the addition of precursor amino acids, on EGT production by the selected strains were also evaluated. U. siamensis showed a 1.5-fold increase in EGT production with the addition of histidine, while U. shanxiensis experienced a 1.8-fold increase in EGT production with the addition of methionine. These results suggest that basidiomycetous yeasts could serve an abundant source for natural EGT producers. [ABSTRACT FROM AUTHOR]

Ergothioneine and selenoneine are structurally related dietary antioxidants and cytoprotectants that may help prevent several chronic diseases associated with inflammation and aging. Both compounds share pharmacokinetic characteristics such as cellular uptake through the ergothioneine transporter, accumulation in red blood cells, and biotransformation to methylated metabolites. A rapid, sensitive, specific, precise, and cost-effective analytical method is required to further investigate the potential health benefits of these compounds, individually or combined, in large epidemiological studies. We developed and validated an isotope-dilution liquid chromatography tandem mass spectrometry (ID-LC–MS/MS) method for the simultaneous specific quantification of these analytes in human blood following a simple sample preparation consisting of dilution in aqueous dithiothreitol followed by centrifugal filtration. Chromatographic separation of the analytes is achieved using a reversed-phase chromatography within an 8-min run. Analyte detection is performed using triple quadrupole mass spectrometry in multiple reaction monitoring mode. Each analyte is quantified against its corresponding isotopically labeled internal standard either commercially available or synthesized in-house (77Se-labeled selenoneine compounds). The validated method demonstrates excellent linearity and very good precision (all CV < 10%). Matrix effects are minimal, suggesting that this method could easily be adapted to other matrices. Freeze/thaw cycles have little effect on methylated metabolites but significantly reduced concentrations of the parent compounds. The method was successfully applied to a small set of volunteer blood samples containing low levels of the analytes. The developed ID-LC–MS/MS method opens new avenues for exploring the roles of these bioactive compounds and their metabolites in human health and disease. [ABSTRACT FROM AUTHOR]

Selenoneine is an organic selenium compound contained in blood and dark muscle of fish. It has a strong antioxidative capacity and is considered useful as a new functional food material. However, the distribution and effects of selenoneine in the mammalian body have not been thoroughly examined. In this study, a selenoneine-rich mackerel extract was developed and fed to mice at 0.07% in standard rodent chow (ME diet) for 32 days to examine its distribution in the body. Selenoneine was distributed in the liver, kidney, and spleen in mice fed with mackerel extract, but it was not distributed in the plasma or erythrocytes. Moreover, concentrations of the major selenium-containing protein were not affected by the mackerel extract. The results of this study suggest that selenoneine is absorbed in the body following ingestion of low doses in crude material and preferentially accumulates in organs and later distributes in erythrocytes. Biochemical analyses of plasma in male mice showed that the glucose level was significantly increased and LDL-cholesterol level was significantly decreased by ME diet feeding. The results indicate that male mice are sensitive to ME diet. [ABSTRACT FROM AUTHOR]

The engineered probiotic Escherichia coli Nissle 1917 (EcN) is expected to be employed in the diagnosis and treatment of various diseases. However, the introduced plasmids typically require antibiotics to maintain genetic stability, and the cryptic plasmids in EcN are usually eliminated to avoid plasmid incompatibility which may change the inherent probiotic characteristics. Here, we provided a simple design to minimize the genetic change of probiotics by eliminating native plasmids and reintroducing the recombinants carrying functional genes. Specific insertion sites in the vectors showed significant differences in the expression of fluorescence proteins. Selected integration sites were applied in the de novo synthesis of salicylic acid, leading to a titer of 142.0 ± 6.0 mg/L in a shake flask with good production stability. Additionally, the design successfully realized the biosynthesis of ergothioneine (45 mg/L) by one-step construction. This work expands the application scope of native cryptic plasmids to the easy construction of functional pathways. Key points: • Cryptic plasmids of EcN were designed to express exogenous genes • Insertion sites with different expression intensities in cryptic plasmids were provided • Target products were stably produced by engineering cryptic plasmids [ABSTRACT FROM AUTHOR]

Potatoes are the most highly consumed vegetable in the United States and are the primary source of antioxidants in the American diet. Therefore, technologies and growing methods that aim to enhance the nutritional quality of potatoes can have positive impacts on public health. Based on past success with other food crops, we hypothesized that inoculation with arbuscular mycorrhizal fungi (AMF) would increase both the yield and nutritional quality of potatoes. To test this hypothesis, we grew yellow fleshed (cv. Lehigh) and purple fleshed (cv. Adirondack Blue) potatoes in containers with several monospecific AMF inoculants comprised of Rhizophagus irregularis, Funneliformis mosseae, or Claroideoglumus etunicatum, and one indigenous mixed species population inoculant. Overall, we found that AMF inoculation increased potato tuber yield by up to 23%, antioxidant activity by up to 120%, ergothioneine concentration by up to 9X, and soluble sugar concentration by up to 46%, and that the extent of these increases varied by mycorrhizal species. Future research should examine the extent to which inoculation with the most beneficial AMF species reported here improves yield and nutritional quality in the field setting. [ABSTRACT FROM AUTHOR]

Ergothioneine (ET) is a naturally occurring antioxidant and cytoprotective agent that is synthesized by fungi and certain bacteria. Recent studies have shown a beneficial effect of ET on neurological functions, including cognition and animal models of depression. The aim of this study is to elucidate a possible effect of ET in rodent models of stroke. Post-ischemic intracerebroventricular (i.c.v.) infusion of ET significantly reduced brain infarct volume by as early as 1 day after infusion in rats, as shown by triphenyltetrazolium chloride (TTC) assay. There was a dose-dependent increase in protection, from 50 to 200 ng of ET infusion. These results suggest that ET could have a protective effect on CNS neurons. We next elucidated the effect of systemic ET on brain infarct volume in mice after stroke. Daily i.p. injection of 35 mg/kg ET (the first dose being administered 3 h after stroke) had no significant effect on infarct volume. However, daily i.p. injections of 70 mg/kg, 100 mg/kg, 125 mg/kg and 150 mg/kg ET, with the first dose administered 3 h after stroke, significantly decreased infarct volume at 7 days after vessel occlusion in mice. In order to elucidate at what time interval during the 7 days there could be effective protection, a second set of experiments was carried out in mice, using one of the effective loading protocols, i.e. 125 mg/kg i.p. ET but the brains were analyzed at 1, 4 and 7 days post-stroke by MRI. We found that ET was already protective against neuronal injury and decreased the size of the brain infarct from as early as 1 day post-stroke. Behavioral experiments carried out on a third set of mice (using 125 mg/kg i.p. ET) showed that this was accompanied by significant improvements in certain behaviors (pole test) at 1 day after stroke. Together, results of this study indicate that i.c.v. and systemic ET are effective in reducing brain infarct volume after stroke in rodent models. [ABSTRACT FROM AUTHOR]

Ergothioneine (ERGO) is a thiol contained in the food that exhibits an excellent antioxidant effect similar to that of glutathione. Although mammals lack a biosynthetic pathway for ERGO, the carnitine/organic cation transporter OCTN1/SLC22A4, which transports ERGO in vivo, is expressed throughout the body, and ERGO is distributed to various organs after oral intake. ERGO is a stable compound that remains in the body for a long time after ingestion. OCTN1 is also expressed in brain parenchymal cells, including neurons, and ERGO in the blood permeates the blood–brain barrier and is distributed to the brain, exhibiting a neuroprotective effect. Recently, the association between central nervous system (CNS) diseases and ERGO has become a research focus. ERGO concentrations in the blood components are lower in patients with cognitive impairment, Parkinson's disease, and frailty than in healthy subjects. ERGO exerts a protective effect against various neurotoxins and improves the symptoms of cognitive impairment, depression, and epilepsy in animal models. The promotion of neurogenesis and induction of neurotrophic factors, in addition to the antioxidant and anti-inflammatory effects, may be involved in the neuroprotective effect of ERGO. This review shows the association between ERGO and CNS diseases, discusses the possible biomarkers of peripheral ERGO in CNS diseases, and the possible preventive and improvement effects of ERGO on CNS diseases. [ABSTRACT FROM AUTHOR]

Exercise is a beneficial practice to the body and can constitute a challenge when taken to the extreme especially in competitive sports. Haematological parameters play a very important role in exercise and are usually good indicators of the effects of exercise on the body. In this experiment, ergothioneine was administered to stallions in the experimental group (Ex + ERG) before exercise to evaluate its ameliorative effect on haematological profile and erythrocytes osmotic fragility against the stress of exercise. The leucocyte count post-exercise in Ex + ERG (5.33 ± 0.24 × 109/L) was lower (P < 0.05) than the count recorded in Ex − ERG which was the untreated group (8.73 ± 0.94 × 109/L). After the exercise, erythrocyte count (15.22 ± 3.74 × 1012/L) recorded in Ex + ERG group was higher (P < 0.05) than counts (12.03 ± 0.68 × 1012/L) obtained in the Ex − ERG group after exercise. Neutrophil/lymphocyte ratio recorded in Ex + ERG group (2.02 ± 0.03) was lower (P < 0.05) than the ratio obtained in Ex − ERG (3.63 ± 0.73). It was concluded that ergothioneine modulated some haematological profiles and decreased leucocyte count and neutrophil/lymphocyte count but increased erythrocyte count and stabilised the membrane of erythrocytes in Arabian stallions subjected to a 2000 m. [ABSTRACT FROM AUTHOR]

Ergothioneine (ET) is a naturally occurring antioxidant that is synthesized by non-yeast fungi and certain bacteria. ET is not synthesized by animals, including humans, but is avidly taken up from the diet, especially from mushrooms. In the current study, we elucidated the effect of ET on the hCMEC/D3 human brain endothelial cell line. Endothelial cells are exposed to high levels of the cholesterol oxidation product, 7-ketocholesterol (7KC), in patients with cardiovascular disease and diabetes, and this process is thought to mediate pathological inflammation. 7KC induces a dose-dependent loss of cell viability and an increase in apoptosis and necrosis in the endothelial cells. A relocalization of the tight junction proteins, zonula occludens-1 (ZO-1) and claudin-5, towards the nucleus of the cells was also observed. These effects were significantly attenuated by ET. In addition, 7KC induces marked increases in the mRNA expression of pro-inflammatory cytokines, IL-1β IL-6, IL-8, TNF-α and cyclooxygenase-2 (COX2), as well as COX2 enzymatic activity, and these were significantly reduced by ET. Moreover, the cytoprotective and anti-inflammatory effects of ET were significantly reduced by co-incubation with an inhibitor of the ET transporter, OCTN1 (VHCL). This shows that ET needs to enter the endothelial cells to have a protective effect and is unlikely to act via extracellular neutralizing of 7KC. The protective effect on inflammation in brain endothelial cells suggests that ET might be useful as a nutraceutical for the prevention or management of neurovascular diseases, such as stroke and vascular dementia. Moreover, the ability of ET to cross the blood-brain barrier could point to its usefulness in combatting 7KC that is produced in the CNS during neuroinflammation, e.g. after excitotoxicity, in chronic neurodegenerative diseases, and possibly COVID-19-related neurologic complications. [ABSTRACT FROM AUTHOR]

Understanding of the underlying mechanism of epilepsy is desired since some patients fail to control their seizures. The carnitine/organic cation transporter OCTN1/SLC22A4 is expressed in brain neurons and transports food-derived antioxidant ergothioneine (ERGO), l-carnitine, and spermine, all of which may be associated with epilepsy. This study aimed to clarify the possible association of this transporter with epileptic seizures. In both pentylenetetrazole (PTZ)-induced acute seizure and kindling models, ocnt1 gene knockout mice (octn1−/−) showed lower seizure scores compared with wild-type mice. Up-regulation of the epilepsy-related genes, c-fos and Arc, and the neurotrophic factor BDNF following PTZ administration was observed in the hippocampus of wild-type, but not octn1−/− mice. To find the OCTN1 substrate associated with the seizure, untargeted metabolomics analysis using liquid chromatography–quadrupole time-of-flight mass spectrometry was conducted on extracts from the hippocampus, frontal cortex, and plasma of both strains, leading to the identification of a plant alkaloid homostachydrine as a compound present in a lower concentration in octn1−/− mice. OCTN1-mediated uptake of deuterium-labeled homostachydrine was confirmed in OCTN1-transfected HEK293 cells, suggesting that this compound is a substrate of OCTN1. Homostachydrine administration increased PTZ-induced acute seizure scores and the expression of Arc in the hippocampus and that of Arc, Egr1, and BDNF in the frontal cortex. Conversely, administration of the OCTN1 substrate/inhibitor ERGO inhibited PTZ-induced kindling and reduced the plasma homostachydrine concentration. Thus, these results suggest that OCTN1 is at least partially associated with PTZ-induced seizures, which is potentially deteriorated by treatment with homostachydrine, a newly identified food-derived OCTN1 substrate. [ABSTRACT FROM AUTHOR]

Brain immune cells, i.e., microglia, play an important role in the maintenance of brain homeostasis, whereas chronic overactivation of microglia is involved in the development of various neurodegenerative disorders. Therefore, the regulation of microglial activation may contribute to their treatment. The aim of the present study was to clarify the functional expression of carnitine/organic cation transporter OCTN1/SLC22A4, which recognizes the naturally occurring food-derived antioxidant ergothioneine (ERGO) as a substrate in vivo, in microglia and its role in regulation of microglial activation. Primary cultured microglia derived from wild-type mice (WT-microglia) and mouse microglial cell line BV2 exhibited time-dependent uptake of [H]- or d9-labeled ERGO. The uptake was markedly decreased in cultured microglia from octn1 gene knockout mice ( octn1 -microglia) and BV2 cells transfected with small interfering RNA targeting the mouse octn1 gene (siOCTN1). These results demonstrate that OCTN1 is functionally expressed in murine microglial cells. Exposure of WT-microglia to ERGO led to a significant decrease in cellular hypertrophy by LPS-stimulation with concomitant attenuation of intracellular reactive oxygen species (ROS), suggesting that OCTN1-mediated ERGO uptake may suppress cellular hypertrophy via the inhibition of ROS production with microglial activation. The expression of mRNA for interleukin-1β (IL-1β) after LPS-treatment was significantly increased in octn1 -microglia and siOCTN1-treated BV2 cells compared to the control cells. Meanwhile, treatment of ERGO minimally affected the induction of IL-1β mRNA by LPS-stimulation in cultured microglia and BV2 cells. Thus, OCTN1 negatively regulated the induction of inflammatory cytokine IL-1β, at least in part, via the transport of unidentified substrates other than ERGO in microglial cells. [ABSTRACT FROM AUTHOR]

Adults with metabolic syndrome from different race/ethnicities are often predisposed to developing type 2 diabetes (T2D); however, growing evidence suggests that healthy diets and lifestyle choices can significantly slow or prevent progression to T2D. This poorly understood relationship to healthy dietary patterns and prevention of T2D motivated us to conduct a retrospective analysis to determine the potential impact of a minor dietary lifestyle change (daily mushroom consumption) on known T2D risk factors in racially diverse adults with confirmed features of the metabolic syndrome. Retrospectively, we studied 37 subjects who had participated in a dietary intervention focused on vitamin D bioavailability from white button mushrooms (WBM). All 37 had previously completed a 16-week study where they consumed 100 g of WBM daily and were then followed-up for one month during which no mushrooms were consumed. We analyzed differences in serum risk factors from baseline to 16-week, and from baseline to one-month follow-up. Measurement of serum diabetic risk factors included inflammatory and oxidative stress markers and the antioxidant component naturally rich in mushrooms, ergothioneine. Significant beneficial health effects were observed at 16-week with the doubling of ergothioneine from baseline, increases in the antioxidant marker ORAC (oxygen radical absorption capacity) and anti-inflammatory hormone, adiponectin and significant decreases in serum oxidative stress inducing factors, carboxymethyllysine (CML) and methylglyoxal (MG), but no change in the lipid oxidative stress marker 8-isoprostane, leptin or measures of insulin resistance or glucose metabolism. We conclude that WBM contain a variety of compounds with potential anti-inflammatory and antioxidant health benefits that can occur with frequent consumption over time in adults predisposed to T2D. Well-controlled studies are needed to confirm these findings and identify the specific mushroom components beneficial to health. [ABSTRACT FROM AUTHOR]

Mushrooms contain a variety of secondary metabolites, including various phenolic compounds and ergothioneine, which have been shown to act as excellent antioxidants. The present study reports a comparison of the antioxidant properties, total phenolic, ergothioneine and mineral contents of the most-consumed strains of Agaricus bisporus (white or brown colors). The samples were fruiting body, mycelia produced at farm (old mycelium) and in vitro (young mycelium). The antioxidant activity was measured by DPPH, FRAP and ABTS assays. The analysis of phenolic compounds was performed by Folin-Ciocalteau method. The mushroom strain with the highest ergothioneine, phenolic compounds and antioxidant potential was the strain A-SPA15 (brown color). Generally, fruiting body samples revealed higher ergothioneine, phenolic compounds and antioxidant properties than mycelia obtained from farm and in vitro culture. There were variations in the mineral content of A. bisporus harvested from the fruiting body, old mycelium, and young mycelium. Furthermore, the lowest heavy-metal concentrations (Ni, Pb, Cd, and Cr) were detected in mycelium samples in each strain. Results demonstrated that mushrooms contained high amounts of copper, zinc, iron and manganese could be used in well-balanced diets. This study contributes to the data relative to A. bisporus consumed as fresh mushrooms and the possibility of in vitro production as a source of bioactive compounds. [ABSTRACT FROM AUTHOR]

The effect of chitosan-ergothioneine coating (CHER) on the post-mortem quality and shelf life of Japanese sea bass ( Lateolabrax japonicas) stored at 4 ± 1 °C for 16 days was investigated. Four different treatments were used: control without coating, CH with chitosan coating, ER with 0.3 % ergothioneine immersion, and CHER with chitosan containing 0.3 % ergothioneine coating. Sea bass pH value, total volatile basic nitrogen, peroxide value, thiobarbituric acid-reactive substances, biogenic amines, color, texture profile, and microbiological and sensory qualities were measured. The results indicate that treatment with CHER coating inhibited increase of total volatile basic nitrogen, peroxide value, and TBA value; maintained tissue hardness; and reduced microorganism counts, such as Pseudomonas, compared with control treatment. The efficiency was better than that of ER or CH treatment. Furthermore, sea bass treated with chitosan-ergothioneine coating also exhibited a positive effect, causing low biogenic amine content, especially putrescine, cadaverine, and histamine. Sensory evaluation proved the efficacy of chitosan-ergothioneine coating by maintaining the overall quality of sea bass during the storage period. Our study suggests that chitosan-ergothioneine coating might be a promising candidate for maintaining sea bass quality and extending their shelf life. [ABSTRACT FROM AUTHOR]

Gastrodia elata is a myco-heterotrophic orchid that has been used in traditional medicine to treat number of human illnesses. It establishes symbiotic associations with compatible mycorrhizal fungi, such as Mycena species and Armillaria mellea, depending on its developmental stages. G. elata tubers contain two major pharmacological substances; gastrodin (GA) and ergothioneine (ERG). Here we investigated whether the use of different tree species could enhance the growth of G. elata tubers, or of the content of GA or ERG. A. mellea infected all 30 tree species but its infection levels varied among species. However, only 14 species stimulated the growth of G. elata immature tubers; Ulmus davidiana produced the biggest tubers (7.8 g FW) while Abies holophylla generated the least significant changes on the growth of tubers reaching their weights to 0.9 g. In contrast to the tuber growth, the infection levels of A. mellea in those trees were similar, indicating that the colonized levels of A. mellea on different tree hosts were not correlated with the yield of G. elata tubers. Interestingly, the weights of immature tubers developed on 14 tree species were negatively correlated with the levels of ERG, but not with those of GA. In conclusion, our study suggested that the use of U. davidiana instead of Quercus species, which has been used for traditional cultivation of G. elata, might enhance the yields of G. elata tubers, but reduce the ERG levels while maintaining the GA levels. [ABSTRACT FROM AUTHOR]

We identified ergothionase, which catalyzes conversion of ergothioneine to thiolurocanic acid and trimethylamine, in a newly isolated ergothioneine-utilizing strain, Burkholderia sp. HME13. The enzyme was purified and its N-terminal amino acid sequence was determined. Based on the amino acid sequence, the gene encoding the enzyme was cloned and expressed in Escherichia coli. The recombinant enzyme was purified to homogeneity and characterized. The enzyme consisted of four identical 55-kDa subunits. The enzyme showed maximum activity at pH 8.0 and 65 °C and was stable between pH 7.0 and pH 10.0 and up to 60 °C. The enzyme acted on ergothioneine ( K: 19 μM, V: 270 μmol/min/mg), but not d-histidine, l-histidine, d-tyrosine, l-tyrosine, d-phenylalanine, or l-phenylalanine. The enzyme was activated by BaCl and strongly inhibited by CuSO, ZnSO, and HgCl. The amino acid sequence of ergothionase showed 23 % similarity to histidine ammonia-lyase (HAL) from Pseudomonas putida and 17 % similarity to phenylalanine ammonia-lyase (PAL) from parsley. However, the tripeptide sequence, Ala-Ser-Gly, which is important for catalysis in both HAL and PAL, was not conserved in ergothionase. The application of ergothionase for the quantification of ergothioneine contained in practical food and blood samples was investigated by performing a recovery test. Satisfactory recovery data (98.7-104 %) were obtained when ergothioneine was added to extract of tamogitake and hemolysis blood. [ABSTRACT FROM AUTHOR]

The native amino acid ergothioneine, a thiourea derivative of histidine, inhibits mushroom tyrosinase activity in a dose-dependent manner, with an IC 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 value for ergothioneine scavenging of 2,2-diphenyl-1-picrylhydrazyl was 6.110 ± 0.305 mg/ml, much higher than the IC 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. [ABSTRACT FROM AUTHOR]

Using 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 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). [ABSTRACT FROM AUTHOR]

Ergothioneine (2-mercaptohistidine trimethylbetaine) is a very stable natural compound largely present in cells and tissues of plants and mammals. The biological role of ET is still incompletely understood but its antioxidant properties are well assessed experimentally. However, no theoretical study had been performed until now to analyze the first steps of the reactions between this natural compound and strong oxidants in order to gain more knowledge on the reaction mechanisms. In this paper, the reaction of ergothioneine with two strong oxidants, hydroxyl radical and peroxinitrite, is modeled in a cluster of forty to fifty water molecules by ab-initio molecular dynamics. The reaction of hydroxyl radical with ergothioneine consists in electron transfer. When hydroxyl radical is initially situated near the sulfur atom, the reaction is not barrier less but passes through a transition state formed by a strong adduct. Otherwise, the charge transfer occurs when the two reactants and a "wire" of water molecules form a loop allowing the charge flow. [ABSTRACT FROM AUTHOR]