Extracellular Peroxidase Activity Research Articles

Eosinophil Specialization Is Regulated by Exposure to the Esophageal Epithelial Microenvironment

Abstract Distinct subsets of eosinophils are reported in inflammatory and healthy tissues, yet the functions of uniquely specialized eosinophils and the signals that elicit them, particularly in eosinophilic esophagitis (EoE), are not well understood. Herein, we report an ex-vivo system wherein freshly isolated human eosinophils were cocultured with esophageal epithelial cells and disease-relevant pro-inflammatory (IL-13) or pro-fibrotic (TGF-β) cytokines. Compared with untreated cocultures, IL-13 increased expression of CD69 on eosinophils, whereas TGF-β increased expression of CD81, CD62L, and CD25. Eosinophils from IL-13–treated cocultures demonstrated increased secretion of GRO-α, IL-8, and M-CSF and also generated increased extracellular peroxidase activity following activation. Eosinophils from TGF-β–treated cocultures secreted increased IL-6 and exhibited increased chemotactic response to CCL11 compared with eosinophils from untreated or IL-13—treated coculture conditions. When eosinophils from TGF-β–treated cocultures were cultured with fibroblasts, they upregulated SERPINE1 expression and fibronectin secretion by fibroblasts compared with eosinophils that were cultured with GM-CSF, alone. Translational studies revealed that CD62L was heterogeneously expressed by eosinophils in patient biopsies. Our results demonstrate that disease-relevant pro-inflammatory and pro-fibrotic signals present in the esophagus of EoE patients cause distinct profiles of eosinophil activation and gene expression.

Putative NAD(P)-Binding Rossmann Fold Protein Is Involved in Chitosan-Induced Peroxidase Activity and Lipoxygenase Expression in Physcomitrium patens.

Oxidative burst, the rapid production of high levels of reactive oxygen species in response to external stimuli, is an early defense reaction against pathogens. The fungal elicitor chitosan causes an oxidative burst in the moss Physcomitrium patens (formerly Physcomitrella patens), mainly due to the peroxidase enzyme Prx34. To better understand the chitosan responses in P. patens, we conducted a screen of part of a P. patens mutant collection to isolate plants with less peroxidase activity than wild-type (WT) plants after chitosan treatment. We isolated a P.patens mutant that affected the gene encoding NAD(P)-binding Rossmann fold protein (hereafter, Rossmann fold protein). Three Rossmann fold protein-knockout (KO) plants (named Rossmann fold KO lines) were generated and used to assess extracellular peroxidase activity and expression of defense-responsive genes, including alternative oxidase, lipoxygenase (LOX), NADPH oxidase, and peroxidase (Prx34) in response to chitosan treatment. Extracellular (apoplastic) peroxidase activity was significantly lower in Rossmann fold KO lines than in WT plants after chitosan treatments. Expression of the LOX gene in Rossmann fold KO plants was significantly lower before and after chitosan treatment when compared with WT. Peroxidase activity assays together with gene expression analyses suggest that the Rossmann fold protein might be an important component of the signaling pathway leading to oxidative burst and basal expression of the LOX gene in P. patens. [Formula: see text] Copyright © 2023 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

Growth, tolerance, and enzyme activities of Trichoderma strains in culture media added with a pyrethroids-based insecticide

The application of pyrethroids and carbamates represents an environmental risk and may exert adverse effects on beneficial microorganisms such as Trichoderma, which contribute to the biocontrol of several fungal phytopathogens. This research evaluated the tolerance of several strains of Trichoderma to a selected culture medium contaminated with a commercial insecticide (H24®) composed of pyrethroids, permethrin and prallethrin, and carbamate propoxur, and determined the influence of this insecticide on the release of enzymes such as chitinases, peroxidases, and endoglucanases by a consortium of selected Trichoderma strains grown in liquid culture medium. Four out of 10 Trichoderma strains showed tolerance to 200ppm (∼48.3% of growth) of the commercial insecticide after 96h of exposure to a contaminated solid medium. After eight days of growth in liquid culture, the insecticide enhanced extracellular protein content and peroxidase activities in the Trichoderma consortium but decreased both chitinase and glucanase activities. These fungal responses should be considered when implementing strategies that combine alternative pesticides and fungal biocontrollers for managing fungal phytopathogens.

Оптимизация ранней диагностики развития и прогрессирования ретинопатии недоношенных

Purpose. To develop easy-to-use methods for early diagnosis of the development and progression of retinopathy of prematurity (ROP). Мaterial and methods. The extracellular peroxidase activity (EPA) of blood serum was studied by spectrophotometry in 93 premature newborns aged 3–4 weeks of life and the thickness of the retinal neuroepithelium (NES) of the paramacular zone by measuring it at a distance of 500 μm from the border of the foveola in 82 premature infants (162 eyes ) using optical coherence tomography (OCT) of the retina on the RetinaScan — 3000 device (NIDEK) in the Macula Multi Cross mode. Results. Determination in premature newborns at 3–4 weeks of life of the level of blood serum VPA less than 1887.1 c.u. allows predicting the development of ROP in 76.4 % of cases. At the same time, there is an inverse correlation (R = –0.94; p < 0.05) between the value of the VPA and the pathology of the upper respiratory tract (respiratory distress syndrome, respiratory failure, asphyxia). The study of the thickness of the paramacular zone of NES showed an increase in its values, regardless of the stage and form of ROP. At the same time, the average thickness index was higher by 39.0 μm in infants with ADHD compared with stage I of the classical form, by 39.3 μm — by stage II, by 35.8 μm — by stage III (p < 0.05). Stabilization of the pathological process was accompanied by a decrease in the NES thickness index (p < 0.05). Conclusion. Examination of the values of extracellular peroxidase activity in the blood serum at 3–4 weeks of life of a premature newborn, as well as monitoring the thickness of the retinal neuroepithelium in the paramacular zone, allow timely diagnosis of retinopathy of prematurity. Keywords: retinopathy of prematurity, diagnostics, extracellular peroxidase activity, retinal neuroepithelial thickness

Enzymatic and Antimicrobial Activities in Polar Strains of Microscopic Soil Fungi.

Potential to produce inducible enzymes (several hydrolases and oxidases) and antibiotics as secondary metabolites was studied in soil micromycete strains from the Arctic (Franz Josef Land and Novaya Zemlya) and Antarctica (the oases Thala Hills, Larsemann Hills, Schirmacher, and Marie Byrd Land). Maximal esterase activity was observed in strains of two typical Antarctic species, Hyphozyma variabilis 218 and Thelebolus ellipsoideus 210 (51 and 29 nmol FDA/((g myceliumh), respectively). Cellulolytic activity was maximal (89 µmol glucose/mg biomass) in Ascochyta pisi 192. Extracellular phenol oxidase (laccase) and peroxidase activities were not detected in the strains examined. Antibacterial activity toward Bacillus subtilis ATCC 6633 was observed in 75% of the Antarctic micromycete strains. Higher-activity strains were isolated from organic-rich moist habitats with a moss or lichen cover. Maximal activities were displayed by Paecilomyces marquandii 166, Penicillium janczewskii 165, Penicillium roseopurpureum 169, and Thelebolus ellipsoideus 210. Antagonistic activity toward Antarctic bacterial strains was shown by 77% of the microfungal strains examined. Maximal inhibition was observed with strains of the typical Antarctic species Antarctomyces psychrotrophicus MT303855 and the eurytopic species Sarocladium kiliense MT303856. Antimycotic activity was observed in 42% of the strains. Both activities were detected in 38% of the Antarctic strains.

Excessive Consumption Hibiscus sabdariffa L. Increases Inflammation and Blood Pressure in Male Wistar Rats via High Antioxidant Capacity: The Preliminary Findings.

Hibiscus sabdariffa L. (HSL) has high amounts of antioxidants and many beneficial effects in several pathologies. However, few studies describe the possible harmful effects of high concentrations of HSL. Here we evaluate the effect of excessive and chronic consumption of infusions with different percentages of HSL on some oxidative stress markers in serum, and the possible association with inflammation and increased systolic blood pressure (SBP), in healthy rats. A total of 32 male Wistar rats were used to form 4 groups with 8 animals each. Group 1 control (drinking tap water), group 2, 3 and 4, drinking water supplemented with 15, 30 and 60 g/L of HSL calyxes respectively. SBP was evaluated and determinations in serum of the NO3−/NO2− ratio, glutathione (GSH), total antioxidant capacity (TAC), selenium (Se), TNF-α, IL-1α/IL-1F1, IL-1β, IL-10, extracellular superoxide dismutase (EcSOD), thioredoxin reductase (TrxR) and glutathione peroxidase (GPx) activities, were evaluated. The SBP (p = 0.01), GPx activity, GSH, TAC, Se, TNF-α and EcSOD activities (p ≤ 0.001) and IL-1α/IL-1F1, IL-1β, TrxR and NO3−/NO2− (p ≤ 0.05), were increased but IL-10 (p < 0.001) was decreased in rats that consumed the 3 and 6% HSL infusions. The excessive and chronic consumption of HSL may increase the TAC that could lead to a proinflammatory state which is associated with hypertension.

Selenium Status and Supplementation Effects in Pregnancy-A Study on Mother-Child Pairs from a Single-Center Cohort.

The demand for selenium (Se) increases during pregnancy since this element supports child growth, proper neuronal development and maternal thyroid function. The issue is particularly relevant for populations living in areas with a limited selenium supply, where many pregnant women opt for Se supplementation. The efficiency of this measure is unknown, although it seems vital in the prevention of severe Se deficiency. In order to evaluate this hypothesis, an observational study was conducted in Poland, where Se deficiency is prevalent. Pregnant women were invited to participate in the study and provided serum samples at the end of pregnancy (n = 115). Information on the supplemental intake of micronutrients was recorded in a face-to-face interview. In addition, serum samples were isolated from the cord blood of newborns at delivery (n = 112) and included in the analyses. Thyroid hormone status was evaluated by routine laboratory tests, and Se status was determined by total Se and selenoprotein P (SELENOP) concentrations and extracellular glutathione peroxidase (GPX3) activity. The three parameters of Se status correlated strongly within the group of mothers and within the group of newborns, with an additional significant correlation found among mother–child pairs. One-third of mothers reported additional Se intake, mainly as a component of multi-micronutrient supplements, at a mean (±SD) dosage of 42 ± 14 µg Se/day. Despite this regime, most of the women presented an insufficient Se status, with 79% of mothers displaying serum Se concentrations below 70 µg/L (indicating Se deficiency) and 22% showing levels below 45.9 µg/L (severe Se deficiency). The inadequate Se supply was also reflected in relatively low SELENOP concentrations and GPX3 activity. Neither total Se nor SELENOP or GPX3 levels were significantly higher in the group of mothers reporting the intake of supplements than in the non-supplementing group. Nevertheless, elevated SELENOP concentrations were observed in the subgroup receiving supplements with more than 55 µg/day. We conclude that the self-administered supplementation of small Se dosages was not sufficient to achieve replete Se status in the micronutrient scant area. However, the maternal Se deficit measured by either Se, SELENOP or GPX3 was transferred from mothers to the newborns, as the parameters correlated strongly in the mother–newborn pairs of samples. It is vital to re-evaluate the guidelines concerning pregnancy care and monitoring of micronutrient status during pregnancy, in particular in areas where deficiencies are present.

NUTRARET: Effect of 2-Year Nutraceutical Supplementation on Redox Status and Visual Function of Patients With Retinitis Pigmentosa: A Randomized, Double-Blind, Placebo-Controlled Trial.

Oxidative stress plays a major role in the pathogenesis of retinitis pigmentosa (RP). The main goal of this study was to evaluate the effect of 2-year nutritional intervention with antioxidant nutraceuticals on the visual function of RP patients. Secondly, we assessed how nutritional intervention affected ocular and systemic redox status. We carried out a randomized, double-blind, placebo-controlled study. Thirty-one patients with RP participated in the study. RP patients randomly received either a mixture of nutraceuticals (NUT) containing folic acid, vitamin B6, vitamin A, zinc, copper, selenium, lutein, and zeaxanthin or placebo daily for 2 years. At baseline and after 2-year of the nutritional supplementation, visual function, dietetic-nutritional evaluations, serum concentration of nutraceuticals, plasma and aqueous humor concentration of several markers of redox status and inflammation were assessed. Retinal function and structure were assessed by multifocal electroretinogram (mfERG), spectral domain-optical coherence tomography (SD-OCT) and automated visual field (VF) tests. Nutritional status was estimated with validated questionnaires. Total antioxidant capacity, extracellular superoxide dismutase (SOD3), catalase (CAT), and glutathione peroxidase (GPx) activities, protein carbonyl adducts (CAR) content, thiobarbituric acid reactive substances (TBARS) formation (as indicator of lipid peroxidation), metabolites of the nitric oxide (NOX) and cytokine (interleukin 6 and tumor necrosis factor alpha) concentrations were assessed by biochemical and immunological techniques in aqueous humor or/and blood. Bayesian approach was performed to determine the probability of an effect. Region of practical equivalence (ROPE) was used. At baseline, Bayesian analysis revealed a high probability of an altered ocular redox status and to a lesser extent systemic redox status in RP patients compared to controls. Twenty-five patients (10 in the treated arm and 15 in the placebo arm) completed the nutritional intervention. After 2 years of supplementation, patients who received NUT presented better retinal responses (mfERG responses) compared to patients who received placebo. Besides, patients who received NUT showed better ocular antioxidant response (SOD3 activity) and lower oxidative damage (CAR) than those who received placebo. This study suggested that long-term NUT supplementation could slow down visual impairment and ameliorate ocular oxidative stress.

Wood-feeding termite gut symbionts as an obscure yet promising source of novel manganese peroxidase-producing oleaginous yeasts intended for azo dye decolorization and biodiesel production

BackgroundThe ability of oxidative enzyme-producing micro-organisms to efficiently valorize organic pollutants is critical in this context. Yeasts are promising enzyme producers with potential applications in waste management, while lipid accumulation offers significant bioenergy production opportunities. The aim of this study was to explore manganese peroxidase-producing oleaginous yeasts inhabiting the guts of wood-feeding termites for azo dye decolorization, tolerating lignocellulose degradation inhibitors, and biodiesel production.ResultsOut of 38 yeast isolates screened from wood-feeding termite gut symbionts, nine isolates exhibited high levels of extracellular manganese peroxidase (MnP) activity ranged between 23 and 27 U/mL after 5 days of incubation in an optimal substrate. Of these MnP-producing yeasts, four strains had lipid accumulation greater than 20% (oleaginous nature), with Meyerozyma caribbica SSA1654 having the highest lipid content (47.25%, w/w). In terms of tolerance to lignocellulose degradation inhibitors, the four MnP-producing oleaginous yeast strains could grow in the presence of furfural, 5-hydroxymethyl furfural, acetic acid, vanillin, and formic acid in the tested range. M. caribbica SSA1654 showed the highest tolerance to furfural (1.0 g/L), 5-hydroxymethyl furfural (2.5 g/L) and vanillin (2.0 g/L). Furthermore, M. caribbica SSA1654 could grow in the presence of 2.5 g/L acetic acid but grew moderately. Furfural and formic acid had a significant inhibitory effect on lipid accumulation by M. caribbica SSA1654, compared to the other lignocellulose degradation inhibitors tested. On the other hand, a new MnP-producing oleaginous yeast consortium designated as NYC-1 was constructed. This consortium demonstrated effective decolorization of all individual azo dyes tested within 24 h, up to a dye concentration of 250 mg/L. The NYC-1 consortium's decolorization performance against Acid Orange 7 (AO7) was investigated under the influence of several parameters, such as temperature, pH, salt concentration, and co-substrates (e.g., carbon, nitrogen, or agricultural wastes). The main physicochemical properties of biodiesel produced by AO7-degraded NYC-1 consortium were estimated and the results were compared to those obtained from international standards.ConclusionThe findings of this study open up a new avenue for using peroxidase-producing oleaginous yeasts inhabiting wood-feeding termite gut symbionts, which hold great promise for the remediation of recalcitrant azo dye wastewater and lignocellulosic biomass for biofuel production.

Features of the inflammatory process of the nasal mucous membrane and paranasal sinuses in patients with chronic rhinosinusitis with nasal polyps

Objectives to assess the features of the inflammatory process of the nasal mucous membrane and sinuses in patients with сhronic rhinosinusitis with nasal polyps (CRSwNP).&#x0D; Material and methods. We analyzed the results of examination of 44 patients with CRSwNP. The objects of investigation were smears-reprints from the mucous membrane of the middle turbinate (rhinocytograms) and the intensity of the extracellular peroxidase activity of the lavage fluids of the maxillary sinuses. The data obtained were processed using the Statistica 8.0 software.&#x0D; Results. When analyzing rhinocytograms of the mucous membrane of the middle turbinates, the epithelial type of rhinocytogram was most often detected in 65.9% of cases (epithelial cells more than 50%); the eosinophilic type in 22.7% (eosinophils more than 1%) and smears with mast cells in 4.6% of cases; the neutrophilic type was less frequent (neutrophils more than 50%) 6.8% of cases. In 34.1% of patients, the cells of the columnar epithelium had pronounced changes in the cytoplasm (vacuolization and enzymatic activity); and in 38.6% of cases the gross pathology was recorded directly in the nucleus of the cells (nucleoli, granules, destruction of the nucleus). The revealed pathology of the cell nucleus suggests the viral etiology of the development of productive inflammation in the nasal cavity and sinuses In the lavage fluids of the maxillary sinuses, a high intensity of extracellular peroxidase activity was recorded (1777.9132.4 a.u. compared to 1225.0190.9 a.u. in patients with chronic purulent rhinosinusitis and 1245.0362.3 a.u. in patients with purulent-polypous rhinosinusitis).&#x0D; Conclusion. Chronic rhinosinusitis with nasal polyps is accompanied by a pronounced inflammatory process of the nasal mucous membrane and paranasal sinuses in the form of structural and functional changes in the cells of the columnar epithelium and a high intensity of extracellular peroxidase activity. The epithelial nucleus damage (the presence of nucleoli, granules, destruction of the nucleus) suggests a viral nature with the development of productive inflammation in the nasal cavity and sinuses.

Biodiversity and biocatalyst activity of culturable hydrocarbonoclastic fungi isolated from Marac\u2013Moruga mud volcano in South Trinidad

Mud volcanoes (MVs) are visible signs of oil and gas reserves present deep beneath land and sea. The Marac MV in Trinidad is the only MV associated with natural hydrocarbon seeps. Petrogenic polyaromatic hydrocarbons (PAHs) in its sediments must undergo biogeochemical cycles of detoxification as they can enter the water table and aquifers threatening ecosystems and biota. Recurrent hydrocarbon seep activity of MVs consolidates the growth of hydrocarbonoclastic fungal communities. Fungi possess advantageous metabolic and ecophysiological features for remediation but are underexplored compared to bacteria. Additionally, indigenous fungi are more efficient at PAH detoxification than commercial/foreign counterparts and remediation strategies remain site-specific. Few studies have focused on hydrocarbonoclastic fungal incidence and potential in MVs, an aspect that has not been explored in Trinidad. This study determined the unique biodiversity of culturable fungi from the Marac MV capable of metabolizing PAHs in vitro and investigated their extracellular peroxidase activity to utilize different substrates ergo their extracellular oxidoreductase activity (> 50% of the strains decolourized of methylene blue dye). Dothideomycetes and Eurotiomycetes (89% combined incidence) were predominantly isolated. ITS rDNA sequence cluster analysis confirmed strain identities. 18 indigenous hydrocarbonoclastic strains not previously reported in the literature and some of which were biosurfactant-producing, were identified. Intra-strain variability was apparent for PAH utilization, oil-tolerance and hydroxylase substrate specificity. Comparatively high levels of extracellular protein were detected for strains that demonstrated low substrate specificity. Halotolerant strains were also recovered which indicated marine-mixed substrata of the MV as a result of deep sea conduits. This work highlighted novel MV fungal strains as potential bioremediators and biocatalysts with a broad industrial applications.

ROS-Dependent Induction of Antioxidant System and Heat Resistance of Wheat Seedlings by Hemin

Effect of 24-h-long treatment with hemin (a donor of carbon monoxide, CO, produced with the participation of heme oxygenase) on heat resistance and the processes of ROS formation and neutralization was investigated in wheat (Triticum aestivum L.) seedlings. It was found that treatment of seedlings with hemin in the concentration range from 0.5 to 10 µM induced an elevation of their resistance to damaging heating (45°C, 10 min). The greatest effect was produced by 5 µM hemin. Hemin treatment was associated with a transient elevation in the roots of extracellular peroxidase activity and a rise in the content of hydrogen peroxide. Elevation of H2O2 content was not observed in the presence of its scavenger dimethyl thiourea (DMTU) and upon treatment of seedlings’ roots with CO absorber hemoglobin and inhibitor of extracellular peroxidase sodium azide; however, this pattern was not followed in the presence of NADPH oxidase inhibitor imidazole. Since decomposition of hemin is associated with the production of redox active Fe2+ ions, the specificity of its action as a CO donor was tested by comparing the effects of hemin and 5 µM FeSO4. At the examined concentration, ferrous sulfate did not affect the generation of hydrogen peroxide in the roots and activity of extracellular peroxidase. In 24 h after the beginning of hemin treatment, activities of superoxide dismutase, catalase, and intracellular peroxidase in the roots rose. Elevation in the activity of antioxidant enzymes induced by hemin was not observed in the presence of DMTU, and treatment of seedling roots with FeSO4 did not modify their activity. Exposure to the CO donor after damaging heating also reduced outflux from the seedling roots of compounds absorbed in the ultraviolet spectral region. Positive influence of hemin on heat resistance of the seedlings was eliminated by CO scavenger hemoglobin and hydrogen peroxide absorber DMTU. Treatment of seedlings with FeSO4 did not alter their survival after heating. Hydrogen peroxide is assumed to participate as a signal mediator in realization of the influence of CO donor on the antioxidant system of wheat seedlings and their heat resistance.

Tracking gene expression, metabolic profiles, and biochemical analysis in the halotolerant basidiomycetous yeast Rhodotorula mucilaginosa EXF-1630 during benzo[a]pyrene and phenanthrene biodegradation under hypersaline conditions.

Polyaromatic phenanthrene (Phe) and benzo[a]pyrene (BaP) are highly toxic, mutagenic, and carcinogenic contaminants widely dispersed in nature, including saline environments. Polyextremotolerant Rhodotorula mucilaginosa EXF-1630, isolated from Arctic sea ice, was grown on a huge concentration range -10 to 500ppm- of Phe and BaP as sole carbon sources at hypersaline conditions (1M NaCl). Selected polycyclic aromatic hydrocarbons (PAHs) supported growth as well as glucose, even at high PAH concentrations. Initially, up to 40% of Phe and BaP were adsorbed, followed by biodegradation, resulting in 80% removal in 10 days. While extracellular laccase, peroxidase, and un-specific peroxygenase activities were not detected, NADPH-cytochrome c reductase activity peaked at 4 days. The successful removal of PAHs and the absence of toxic metabolites were confirmed by toxicological tests on moss Physcomitrium patens, bacterium Aliivibrio fischeri, human erythrocytes, and pulmonary epithelial cells (A549). Metabolic profiles were determined at the midpoint of the biodegradation exponential phase, with added Phe and BaP (100ppm) and 1M NaCl. Different hydroxylated products were found in the culture medium, while the conjugative metabolite 1-phenanthryl-β-D-glucopyranose was detected in the medium and in the cells. Transcriptome analysis resulted in 870 upregulated and 2,288 downregulated transcripts on PAHs, in comparison to glucose. Genomic mining of 61 available yeast genomes showed a widespread distribution of 31 xenobiotic degradation pathways in different yeast lineages. Two distributions with similar metabolic capacities included black yeasts and mainly members of the Sporidiobolaceae family (including EXF-1630), respectively. This is the first work describing a metabolic profile and transcriptomic analysis of PAH degradation by yeast.

Calcium-Dependent Changes in Cellular Redox Homeostasis and Heat Resistance of Wheat Plantlets under Influence of Hemin (Carbon Monoxide Donor)

Carbon monoxide (CO) is considered to be an important molecule-gasotransmitter involved in the regulation of the functional activity of plants, including in the processes of adaptation to stress factors. However, the associations of CO with other participants of signaling in the plant cells remain poorly studied. Using an inhibitory method, the role of different calcium pools in realization of the influence of hemin (carbon monoxide donor) on generation and neutralization of reactive oxygen species (ROS) in the root cells of wheat (Triticum aestivum L.) plantlets and their resistance to damaging heating (45°C, 10 min) was studied. The treatment of plantlets with 5 μM hemin caused a transient increase in the activity of extracellular peroxidase in roots and an increase in the generation of ROS with the maximum in 1.5–2 h after the beginning of treatment. The chelator of extracellular calcium EGTA and the inhibitor of inositol-1,4,5-phosphate formation neomycin, which reduces calcium influx into the cytosol from intracellular compartments, almost completely eliminated an increase in the activity of extracellular peroxidase caused by exogenous CO. At the same time, EGTA (completely) and neomycin (partially) leveled an increase in the content of hydrogen peroxide in the roots of plantlets occurring under the influence of CO donor. The treatment of plantlets with hemin also induced an increase in the activity of superoxide dismutase, catalase, and intracellular peroxidase in roots. Both calcium antagonists eliminated these effects. In the presence of EGTA and neomycin, there was also no positive effect of the treatment of hemin on the state of biomembranes and survival of plantlets after the damaging heating. It was concluded that both extracellular calcium and that deposited in intracellular compartments is involved in CO donor-induced increase in the formation of ROS, induction of the antioxidant system, and development of heat resistance of wheat plantlets.

Neutrophil activation by Escherichia coli isolates from human intestine: effects of bacterial hydroperoxidase activity and surface hydrophobicity.

Successful colonization of the intestine requires that bacteria interact with the innate immune system and, in particular, neutrophils. Progression of inflammatory bowel diseases (IBD) is associated with alterations in gut microbiota, and dysbiosis in Crohn’s disease (CD) patients is often associated with an expansion of Escherichia coli. Here, we investigated the ability of such E. coli isolates to avoid neutrophil activation and to utilize reactive oxygen species. Neutrophil activation was detected in vitro in normal human blood via luminol chemiluminescence (CL) induced by reactive oxygen and halogen species generated by neutrophils. No significant difference in neutrophil activation in vitro was detected between isolates from inflamed (23 isolates) vs healthy intestines (5 isolates), with 10‐fold variation within both groups (2.9–61.2 mV). CL activity of isolates from the same patient differed by 1.5–5 times. Twenty‐four isolates from ileal aspirate, biopsy, and feces of seven patients with CD and one patient with no intestine inflammation were tested for extracellular peroxidase and catalase activity and cell surface hydrophobicity. Average values between patients varied from 26 ± 3 to 73 ± 18 µmol·g−1 of air dry weight for peroxidase activity, from 15 ± 2 to 189 ± 56 mmol·g−1 of air dry weight for catalase activity, and from 5 ± 3 to 105 ± 9 a.u. for the hydrophobic probe fluorescence. Extracellular peroxidase activity and hydrophobicity of bacterial cell surface correlated negatively with stimulated neutrophil CL. The ability of some isolates to avoid neutrophil activation and to utilize reactive oxygen species may provide a strategy to survive assault by the innate immune system.

Fungal elimination of toluene vapor in one- and two-liquid phase biotrickling filters: Effects of inlet concentration, operating temperature, and peroxidase enzyme activity

In this study, performance of biotrickling filters (BTFs) inoculated with fungus Phanerochaete chrysosporium at 30 °C and 40 °C in the absence and presence of silicone oil (10% v/v) was investigated. Removal of toluene was carried out at empty bed residence time (EBRT) of 1 min and at inlet concentrations of 0.5–4.4 g m−3 and 0.5–24.7 g m−3 for one-liquid phase (OLP-BTF) and two-liquid phase BTF (TLP-BTF), respectively. In general, at 40 °C, removal efficiencies (REs) > 80% were obtained in OLP-BTF for the inlet toluene concentrations < 2.5 g m−3, and REs > 70% were obtained for concentrations < 18 g m−3 in TLP-BTF. Based on the balanced equation for biodegradation, fungal respiration produced more CO2 in OLP-BTF (1.38 mol CO2/mole toluene) in comparison to TLP-BTF (0.67 mol CO2/mole toluene). In other words, the presence of oil enhanced microbial growth due to the increase of hydrophobic substrate bioavailability. The activity of extracellular ligninolytic manganese peroxidase (MnP) enzyme produced by the fungal culture was detected in the range of 27.6–71.6 U L−1 (μmol min−1 L−1) at 40 °C in TLP-BTF, while no enzymatic activity was detected in OLP-BTF.

Ash application enhances decomposition of recalcitrant organic matter

Harvesting whole-tree biomass for biofuel combustion intensifies removal of nutrients from the ecosystem. This can be partly abated by applying ash from the combustion back to the system, as the ash is rich in nutrients. Ash is very alkaline and ash application raises soil pH, which in turn can stimulate microbial activity and thus decomposition and mineralization. Our aim was to test if ash induced decomposition activity was associated with enhanced turnover of recalcitrant, i.e. relatively old, organic pools. Two experiments were conducted in the same coniferous plantation after the application of 0, 3, 4.5 and 6 t ash ha−1, and 0, 3, 9, 15 and 30 t ash ha−1, respectively. We used natural abundance of 15N in mosses, mites and ectomycorrhizal fungi 26 months after ash application, as well as temporal variation in δ15N values of ectomycorrhizal fungi, as an indicator of decomposition of recalcitrant organic matter in the first experiment. Furthermore, in the second experiment we used measurements of extracellular manganese peroxidase activity almost 4 years after ash application as an indication of potential decomposition of lignin, an important component of recalcitrant organic matter.The δ15N signature increased significantly for ectomycorrhizal fungi, dead moss, Nothroid and Gamasida mites, and manganese peroxidase activity tended to increase, with increasing ash doses. This suggests that ash application stimulates turnover of recalcitrant organic matter, which can increase the available pool of nitrogen in the system. This will potentially enhance the fertilizer value of ash. However, the δ15N in ectomycorrhizal fungi tended to peak at 18 months after ash application, before decreasing, suggesting that the turnover of recalcitrant organic matter is reduced again with time.

Characterization of the Papain-Like Protease p29 of the Hypovirus CHV1-CN280 in Its Natural Host Fungus Cryphonectria parasitica and Nonhost Fungus Magnaporthe oryzae.

Cryphonectria hypovirus 1 strain CN280 (CHV1-CN280) was isolated from North China and exhibited typical hypovirulence-associated traits. We previously reported that CHV1-CN280 was more aggressive and had a higher horizontal transmission ability between Cryphonectria parasitica isolates belonging to different vegetative compatibility groups than two other CHV1 hypoviruses (namely, CHV1-EP713 and CHV1-Euro7), thus displaying greater potential for biological control of chestnut blight. The genome sequence of CHV1-CN280 shared approximately 70% identity with three other hypoviruses (CHV1-EP713, CHV1-Euro7, and CHV1-EP721). The coding region for p29, a papain-like protease encoded by CHV1-CN280 hypovirus, displayed an average of only approximately 60% amino acid identity among them, while the identity between the other three CHV1 isolates was higher than 89%. Protease p29 acted as a virus-encoded determinant responsible for altering fungal host phenotypes in other CHV1 isolates. In this study, the impacts of CHV1-CN280 p29 expression in virus-free C. parasitica were investigated. CHV1-CN280 p29 expression in C. parasitica resulted in significantly reduced sporulation, pigmentation, extracellular laccase activities, and pathogenicity, which is consistent with previous investigations. Subsequently, the potential of CHV1-CN280 p29 as a viral determinant responsible for suppression of host phenotypes in other phytopathogenic fungi such as Magnaporthe oryzae, the causal agent of rice blast disease, was discussed. However, heterologous expression of p29 in M. oryzae induced the opposite effect on sporulation, extracellular laccase activities, and pathogenicity; had no significant effect on pigmentation and mycelial growth; and contributed to extracellular peroxidase activities, suggesting that CHV1-CN280 p29 may disturb a unique regulatory pathway in C. parasitica, rather than a basic regulatory pathway conserved in diverse range of fungi. Alternatively, CHV1-CN280 p29-mediated modulation of fungal phenotypes may be facilitated by the specific interaction between p29 and a special fungal-host component, which exists only with C. parasitica but not M. oryzae.

Agrowastes utilization by Raoultella ornithinolytica for optimal extracellular peroxidase activity.

The industrial applications and prospects of microbial peroxidase are on the upwards trend, thus necessitating the search for sources with high turnaround time. Actinobacterial species have been a major source of peroxidase for the obvious reasons of having robust metabolite expression capabilities. However, other bacteria species have been underexplored for peroxidase production, hence the motivation for the investigation into the peroxidase production potential of Raoultella ornithinolytica OKOH-1 (KX640917). The bacteria expressed optimum specific peroxidase activity of 16.48 ± 0.89Umg-1 , which is higher than those previously reported. The optimal fermentation conditions were pH 5 (3.44 ± 0.64UmL-1 ), incubation temperature of 35 °C (5.25 ± 0.00UmL-1 ), and agitation speed of 150rpm (9.45 ± 2.57UmL-1 ), with guaiacol and ammonium chloride as the best inducer and nitrogen supplement, respectively. On valorization of agrowastes as a sole carbon source for the secretion of peroxidase, sawdust gave the best peroxidase yield (15.21 ± 2.48Umg-1 ) under solid-state fermentation. Also, a nonperoxide-dependent enzyme activity, which suggests probable laccase activity, was observed. The ability of the bacteria to utilize agrowastes is highly economical and as well a suitable waste management strategy. Consequently, R. ornithinolytica OKOH-1 is a promising industrial strain with dexterity for enhanced peroxidase production.

Down-regulation of pyruvate decarboxylase gene of white-rot fungus Phlebia sp. MG-60 modify the metabolism of sugars and productivity of extracellular peroxidase activity

Ethanologenic white-rot fungus Phlebia sp. MG-60-P2 produces ethanol directly from several lignocelluloses. Efficient gene silencing methods are needed for metabolic engineering of this fungus for biorefinery use. In this study, we evaluated the effectiveness of RNAi-mediated silencing of the pyruvate decarboxylase gene of Phlebia sp. MG-60-P2 (MGpdc1). The RNAi lines generated showed a variety of suppression levels of ethanol production and MGpdc1 expression, and two selected strains led to different metabolic fluxes, resulting in rapid accumulation of xylitol from xylose. Knockdown lines KD2 and KD10 showed different strength of silencing. The moderate-inhibition line (KD10) showed faster xylitol accumulation from xylose than the severe-inhibition line (KD2). KD2, KD10 and knockout line KO77 showed higher extracellular peroxidase activity than the wild-type. Gene silencing using RNAi for MGpdc1 in the ethanologenic white-rot fungus Phlebia sp. MG-60-P2 is an effective first step in metabolic engineering to produce other chemicals besides ethanol. This high efficiency of transformation and silencing effect will make it possible to cotransform with multiple expression vectors which enhance the minor metabolic pathway or introduce exogenous metabolic reaction in Phlebia sp. MG-60-P2.