Comparison of the potential of Ficus sycomorus latex and horseradish peroxidases in the decolorization of synthetic and natural dyes

The aim of this study was to compare the potential of Ficus sycomorus latex peroxidase (POL) and horseradish peroxidase (HRP) in the decolorization of a wide spectrum of eight synthetic dyes and two natural dyes, hibiscus flower color and pomegranate juice. We study for the first time the decolorization of natural dyes enzymatically. The highest decolorization percent was reported at 20 mg/l for all dyes treated with POL and HRP. Both the enzymes had lower decolorization % for azo-carmin (30–33%). During decolorization treatment, both natural dyes and titan yellow formed precipitates which settled down and were removed by centrifugation. The enhancement of the decolorization % of the most tested dyes by treatment with POL and HRP was reported in the presence of some redox mediators. The rate of decolorization was enhanced by increasing the time and the most significant changes were observed during the first 6 h of incubation. One hundred percent enhancement in decolorization was reported for azo-carmine in the presence of histidine and α-naphthol as redox mediators. A few of redox mediators caused no significant effect or decreases the decolorization % for a little number of tested dyes. The decolorization of dyes by POL and HRP in the presence of redox mediators appeared without the formation of precipitate. A similar decolorization % for all the tested dyes by POL and HRP was detected. The data suggested that the peroxidase/mediator system was an effective biocatalyst for the decolorization of synthetic and natural dyes, and POL could be used as a potential option for the application of dye decolorization.     

Coupled electron and proton transfer reactions during the O → E transition in bovine cytochrome c oxidase

A combined DFT/electrostatic approach is employed to study the coupling of proton and electron transfer reactions in cytochrome c oxidase (CcO) and its proton pumping mechanism. The coupling of the chemical proton to the internal electron transfer within the binuclear center is examined for the O → E transition. The novel features of the His291 pumping model are proposed, which involve timely well-synchronized sequence of the proton-coupled electron transfer reactions. The obtained pK_as and E_ms of the key ionizable and redox-active groups at the different stages of the O → E transition are consistent with available experimental data. The PT step from E242 to H291 is examined in detail for various redox states of the hemes and various conformations of E242 side-chain. Redox potential calculations of the successive steps in the reaction cycle during the O → E transition are able to explain a cascade of equilibria between the different intermediate states and electron redistribution between the metal centers during the course of the catalytic activity. All four electrometric phases are discussed in the light of the obtained results, providing a robust support for the His291 model of proton pumping in CcO. This article is part of a Special Issue entitled: Respiratory oxidases.     

Molecular analysis of the bacterial community in digestive tract of rabbit

This work aimed to study the stability over time of the bacterial community in cæcum and fæces of the rabbit (diversity index and structure) without experimental disturbance and to evaluate its relationships with environmental parameters. Soft and hard fæces of 14 rabbits were sampled for 5 weeks while cæcal content was sampled on the 3rd week (by surgery) and the 5th week (at slaughter). Bacterial communities were assessed by studying CE-SSCP profiles of 16S rRNA genes fragments. Redox potential, pH, NH3-N concentration and volatile fatty acid concentrations were measured in the cæcum. Data showed that bacterial communities of soft and hard fæces barely differed from that of the cæcum (ANOSIM-R < 0.25; p < 0.05). Without disturbance, the bacterial communities of fæces were stable over time (ANOSIM-R < 0.25; p < 0.001). However, the bacterial communities of cæcum and fæces were affected by the surgery (ANOSIM-R = 0.22–0.33; p < 0.001). The cæcal content was an acidic (pH = 6.03 ± 0.33) and an anaerobic environment (redox potential = ?160 ± 43 mV). Only the redox potential was correlated with the diversity index of the bacterial community of the cæcum (R² = 0.35; p < 0.05) and no environmental parameters were correlated to its structure.     

Correlations between reduction–oxidation potential profiles and growth patterns of Saccharomyces cerevisiae during very-high-gravity fermentation

When Saccharomyces cerevisiae was grown under three glucose concentrations (ca. 200, 250, and 300 g/l), controlled at three reduction–oxidation (redox) potentials (no control, −150 and −100 mV) by manipulating two aerations (0.82 and 1.3 vvm), we observed that the recorded redox potential profiles resembled bathtub curves, and the profiles correlated well to the growth patterns measured under the same conditions. According to the shape of bathtub curve, we subdivided the curve into four regions. Region I features an abrupt decline in redox potential (corresponding to the growth phase from lag and logarithmic to the onset of stationary phase) that correlates to rapid yeast propagation, resulting from fast glucose uptake. Region II (corresponding to the stationary phase in yeast growth, characterizes a constant level of redox potential) is maintained by proper sparging and constant agitation. The continual buildup of ethanol causes growth arrest of yeast, resulting in the reduction of net NADH production. As a result, an uprising of redox potential is the feature of Region III, which signifies the end of stationary phase followed by the commencement of death phase. The severity of growth environment due to ethanol toxicity results in a rapid decrease in yeast population. Region IV (corresponding to the death phase during yeast growth) characterizes a drastic reduction in yeast viability and a gradual leveling of redox potential. A low glucose feed correlates to a fast decline of redox potential, a small basin in the bathtub curve, short fermentation duration, and complete glucose utilization. Imposing the current redox potential settings to low glucose feeds exerts no appreciable effect on ethanol production. In contrast, a high glucose feed connects to a sluggish bathtub curve for all four regions and incomplete glucose utilization. Proximate analysis on carbon balance indicates that controlling redox potential at −150 mV and under ca. 250 and 300 g glucose/l conditions, gave the highest fermentation efficiency as compared to other conditions; but there were no beneficiary effect to control redox potential under ca. 200 g glucose/l conditions.     

Effects of competition outcome on testosterone concentrations in humans: An updated meta-analysis

A contribution to a special issue on Hormones and Human Competition.Since Archer's (2006) influential meta-analysis, there has been a major increase in the number of studies investigating the effect of competition outcome on testosterone reactivity patterns in humans. Despite this increased research output, there remains debate as to whether competition outcome modulates testosterone concentrations. The present paper examines this question using a meta-analytic approach including papers published over the last 35 years. Moreover, it provides the first meta-analytic estimate of the effect of competition outcome on testosterone concentrations in women. Results from a meta-analysis involving 60 effect sizes and > 2500 participants indicated that winners of a competition demonstrated a larger increase in testosterone concentrations relative to losers (D = 0.20)—an effect that was highly heterogeneous. This ‘winner-loser’ effect was most robust in studies conducted outside the lab (e.g., in sport venues) (D = 0.43); for studies conducted in the lab, the effect of competition outcome on testosterone reactivity patterns was relatively weak (D = 0.08), and only found in studies of men (D = 0.15; in women: D = − 0.04). Further, the 'winner-loser' effect was stronger among studies in which pre-competition testosterone was sampled earlier than (D = 0.38, after trim and fill correction) rather than within (D = 0.09) 10 min of the start of the competition. Therefore, these results also provide important insight regarding study design and methodology, and will be a valuable resource for researchers conducting subsequent studies on the 'winner loser' effect.     

Peroxidase-like activity of cytochrome b5 is triggered upon hemichrome formation in alkaline pH

In alkaline media (pH 12) a catalytic peroxidase activity of cytochrome b₅ was found associated to a different conformational state. Upon incubation at this pH, cytochrome b₅ electronic absorption spectrum was altered, with disappearance of characteristic bands of cytochrome b₅ at pH 7.0. The appearance of new electronic absorption bands and EPR measurements support the formation of a cytochrome b₅ class B hemichrome with an acquired ability to bind polar ligands. This hemichrome is characterized by a negative formal redox potential and the same folding properties than cytochrome b₅ at pH 7. The acquired peroxidase-like activity of cytochrome b₅ found at pH 12, driven by a hemichrome formation, suggests a role of this protein in peroxidation products propagation.     

Uprising the antioxidant power of Argania spinosa L. callus through abiotic elicitation

This study was carried out in order to investigate the ability of tissues of Argania spinosa (L.) to undergo unlimited cell divisions by triggering their proliferative potential via callogenesis. Axenic cultures were efficiently established using axillary buds cultured on half-strength Murashige and Skoog (MS) medium after 20 min of surface sterilization with sodium hypochlorite 6% (v/v). The highest callus rate was achieved with 1.0 mg L⁻¹ of naphthaleneacetic acid (NAA) and 1.0 mg L⁻¹ of 2,4-dichlorophenoxyacetic acid (2,4D) or similarly with 0.01 mg L⁻¹ of 6-benzylaminopurine (BAP) and 1.0 mg L⁻¹ of 2,4D at pH of 5.8, under dark conditions. The results of this study show also a significant increase in the callus's antioxidant power under abiotic pressure induced by NaCl. Catalase (CAT), peroxidase (PO), and superoxide dismutase (SOD) activities were significantly triggered, which protected the cells from the stimulated oxidative stress, under hydrogen peroxide (H₂O₂) significant release. This reaction favors subsequently the tissue recover process linked to the low abundance of polyphenol oxidase (PPO) activity and malondialdehyde (MDA) content. This work proves the efficiency of salt stress in boosting the argan cell's antioxidant status, which could be commercially applied in the field of cells regenerative therapy.     

The historical dendroarchaeology of two log structures at the Wynnewood State Historic Site,Castalian Springs,Tennessee, USA

Tree rings of eastern red cedar (Juniperus virginiana L.) were examined from cores extracted from two log cabins located at the Wynnewood State Historic Site in Castalian Springs, Sumner County, Tennessee. One cabin was reportedly built by the first explorer in the area, Isaac Bledsoe, sometime between 1772 and his death in 1793. The second cabin was known as Spencer's Cabin after the first settler of the region, Thomas Sharp Spencer, who lived in the immediate vicinity from 1776 to 1779. The goal of this research was to determine the probable construction year(s) for both cabins and determine whether Bledsoe and Spencer did indeed build these structures. Forty-one cores were extracted from Bledsoe's Cabin, and 30 were used for crossdating and building a floating chronology using COFECHA. The Bledsoe's Cabin chronology was then statistically and graphically crossdated using the eastern red cedar reference tree-ring chronology (ITRDB #TN031) from Norris Dam, Tennessee. We found a statistically significant correlation (r = 0.42, t = 4.18, n = 85, p < 0.0001) between the Bledsoe's Cabin chronology and the reference chronology, anchoring the chronology between 1720 and 1804, with nearly all cores indicating tree harvesting between February and April 1805. Twenty-two cores were extracted from Spencer's Cabin, and 17 were used to build a floating chronology for the cabin. Again, we found a statistically significant correlation (r = 0.44, t = 4.85, n = 100, p < 0.00001) with the reference chronology, which anchored the Spencer's Cabin chronology between 1726 and 1825. All trees appear to have been harvested between February and August in 1826. Therefore, neither structure was built by its historical namesake. No known historical documents suggest who the potential builders were, although the property was owned between ca. 1797 and 1829 by General James Winchester. He and his family, however, never resided on the Wynnewood property because Winchester had built a large multi-room structure in nearby Gallatin, Tennessee, by 1802.     

Evaluation and comparison of four methods of ranking horses based on reactivity

Four methods of ranking horses on reactivity were evaluated and compared: isolation from conspecifics, presentation of a static novel stimulus, traversing a novel stimulus in a runway (isolation, novel stimulus and runway tests, respectively) and assigning subjective emotionality scores. In all tests, horses’ heart rates were recorded and behaviour was videotaped. To be considered a valid test of reactivity, at least one heart rate and one behavioural measurement in the test had to change significantly between treatments (tranquilizer administation versus sham tranquilizer administration), and behavioural measures had to be displayed in at least 75% of the trials. Forty horses performed each of the three tests daily on three different days in a switchback design. Horses were assigned randomly to a daily test sequence, which was maintained throughout the study. In the runway test, no significant difference in heart rate values in tranquilized and non-tranquilized horses was found, and no behavioural attribute was displayed in more than 52% of the trials; therefore it was rejected as a valid test of reactivity. Both isolation and novel stimulus tests produced valid measurements. Mean heart rate was the most precise physiological measure for these tests, and walking and defecation frequency were the most precise behavioural measures for novel stimulus and isolation tests, respectively. Mean heart rates on the novel stimulus and isolation tests were correlated (r_s = 0.79, P < 0.01) indicating that these tests produced similar rankings based on physiological responses. However, behavioural measures ranked horses differently (r_s = 0.27, P < 0.10) on the tests. Rank correlations between mean heart rates and behavioural measures were higher in the novel stimulus (r_s = 0.66, P < 0.01) than the isolation test (r_s = 0.55, P < 0.01), indicating that the novel stimulus test ranked horses based on either physiological or behavioural responses more similarly than did the isolation test. Therefore, the novel stimulus test was considered the more accurate evaluation of reactivity. Subjective emotionality scores were correlated moderately with mean heart rates (r_s > 0.33, P < 0.01) from the novel stimulus and isolation tests and with walking scores (r_s = 0.47, P < 0.01) from the novel stimulus test. Assignment of subjective emotionality scores was not as accurate as the novel stimulus or isolation tests in ranking horses for reactivity. Using physiological data alone, combining physiological and behavioural measurements or using more than one behavioural measurement in reactivity tests may reflect the reactivity of the horse better than a single behavioural measurement.     

Quaternary structure of KATP channel SUR2A nucleotide binding domains resolved by synchrotron radiation X-ray scattering

Heterodimeric nucleotide binding domains NBD1/NBD2 distinguish the ATP-binding cassette protein SUR2A, a recognized regulatory subunit of cardiac ATP-sensitive K⁺ (K_ATP) channels. The tandem function of these core domains ensures metabolism-dependent gating of the Kir6.2 channel pore, yet their structural arrangement has not been resolved. Here, purified monodisperse and interference-free recombinant particles were subjected to synchrotron radiation small-angle X-ray scattering (SAXS) in solution. Intensity function analysis of SAXS profiles resolved NBD1 and NBD2 as octamers. Implemented by ab initio simulated annealing, shape determination prioritized an oblong envelope wrapping NBD1 and NBD2 with respective dimensions of 168 × 80 × 37 Å³ and 175 × 81 × 37 Å³ based on symmetry constraints, validated by atomic force microscopy. Docking crystal structure homology models against SAXS data reconstructed the NBD ensemble surrounding an inner cleft suitable for Kir6.2 insertion. Human heart disease-associated mutations introduced in silico verified the criticality of the mapped protein–protein interface. The resolved quaternary structure delineates thereby a macromolecular arrangement of K_ATP channel SUR2A regulatory domains.     

Application of synthetic peptides for detection of anti-citrullinated peptide antibodies

Anti-citrullinated protein antibodies (ACPAs) are a hallmark of rheumatoid arthritis (RA) and represent an important tool for the serological diagnosis of RA.In this study, we describe ACPA reactivity to overlapping citrullinated Epstein-Barr virus nuclear antigen-1 (EBNA-1)-derived peptides and analyze their potential as substrates for ACPA detection by streptavidin capture enzyme-linked immunosorbent assay. Using systematically overlapping peptides, containing a 10 amino acid overlap, labelled with biotin C-terminally or N-terminally, sera from 160 individuals (RA sera (n = 60), healthy controls (n = 40), systemic lupus erythematosus (n = 20), Sjögren’s syndrome (n = 40)) were screened for antibody reactivity.Antibodies to a panel of five citrullinated EBNA-1 peptides were found in 67% of RA sera, exclusively of the IgG isotype, while 53% of the patient sera reacted with a single peptide, ARGGSRERARGRGRG-Cit-GEKR, accounting for more than half of the ACPA reactivity alone. Moreover, these antibodies were detected in 10% of CCP2-negative RA sera. In addition, 47% of the RA sera reacted with two or three citrullinated EBNA-1 peptides from the selected peptide panel. Furthermore, a negative correlation between the biotin attachment site and the location of citrulline in the peptides was found, i.e. the closer the citrulline was located to biotin, the lower the antibody reactivity.Our data suggest that citrullinated EBNA-1 peptides may be considered a substrate for the detection of ACPAs and that the presence of Epstein-Barr virus may play a role in the induction of these autoantibodies.     

Potential of Typha angustifolia for phytoremediation of heavy metals from aqueous solution of phenol and melanoidin

Typha angustifolia was evaluated for various heavy metals (Cu, Pb, Ni, Fe, Mn, and Zn) bioremediation potential from aqueous solution containing variable concentrations of phenol (100–800 mg l^?1) and melanoidin (2500–8500 Co–Pt) at 20, 40, and 60 days. The concentration of phenol (200–400 mg l^?1) along with melanoidin 2500 Co–Pt showed optimum for phytoremediation of tested heavy metals, while, higher concentrations of melanoidin (5600–8500 Co–Pt) showed toxic effect on T. angustifolia along with phenol. Phenol and melanoidin showed adverse effect on T. angustifolia of up to 20 days incubation, but this leads to induction of peroxidase and ascorbic acid activity to cope with adverse conditions. Subsequently, as pollutants were decreased along with plant growth, peroxidase and ascorbic acid also declined. However, with reduction of peroxidase, catalase level was increased. The Cu, Zn, and Ni were accumulated at maximum in all tested conditions. The TEM observations of T. angustifolia showed clotted deposition of metals and shrinkage of cell in root, breakdown of spongy and palisade parenchyma of leaves at higher concentration of phenol (100 mg l^?1) and melanoidin (5500 Co–Pt). Thus, this study concluded that T. angustifolia could be a potential phytoremediator for heavy metals from metal, melanoidin, and phenol containing industrial wastewater at optimized condition.     

Responsiveness of dairy cows to human approach and novel stimuli

This study investigated intra-test and inter-test consistency of dairy cattle behavioural responses to a series of tests involving human approach and exposure to novelty. Thirty-six lactating Holstein–Friesian cows were each subjected to three human approach tests and three novel stimuli tests. Flight response score was assessed by an experimenter approaching cows when they were in the passageway of the home-pen (AP), lying down (AL) and at the feedface (FF). Each human approach test was repeated on each cow three times. The intra-animal repeatability estimates were 0.65, 0.40, 0.27 for AP, AL and FF tests, respectively. Repeatability evaluates an individual's consistency across tests by comparing it to the variation within the group. Cows showed moderate consistency in their flight response scores to the different approach tests (W₃₅ = 0.56, P < 0.01). Three novel stimuli (water spray, striped boards and flashing light) were individually presented once to each cow. Investigatory and reactivity behavioural responses were assessed. Cows showed the greatest reactivity response to the water spray compared to the striped boards (U₁ = 56, P < 0.001) and flashing light (U₁ = 66, P < 0.001). No statistically significant agreement existed between the novel stimuli reactivity and investigatory responses with the AP flight response scores. In conclusion, consistency over time was demonstrated over a relatively short period for the AP test and consistency between human approach situations was shown, however, consistency between human and novel situations was not found.     

Kinetic analysis for suicide-substrate inactivation of microperoxidase-11: A modified model for bisubstrate enzymes in the presence of reversible inhibitors

Kinetics of microperoxidase-11 (MP-11) as a heme–peptide enzyme model in oxidation reaction of guaiacol (AH) by hydrogen peroxide was studied in the presence of amino acids, taking into account the inactivation of MP-11 during reaction by its suicide substrate, H₂O₂. Reliability of the kinetic equation was evaluated by non-linear mathematical fitting. Fitting of experimental data into a new integrated kinetic relation showed a close match between the kinetic model and the experimental data. Indeed, it was found that the mechanism of suicide-peroxide inactivation of MP-11 in the presence of amino acids is different from MP-11 and/or horseradish peroxidase. In this mechanism, amino acids compete with hydrogen peroxide for the sixth co-ordination position of iron atom in the heme group through a competitive inhibition mechanism.The proposed model can successfully determine the kinetic parameters including inactivation by hydrogen peroxide as well as the inhibitory rate constants by the amino acid inhibitor.Kinetic parameters of inactivation including the initial activity of MP-11, α₀, the apparent inactivation rate constant, k_i and the apparent inhibition rate constant for cysteine, k_I were obtained 0.282 ± 0.006 min^?1, 0.497 ± 0.013 min^?1 and 1.374 ± 0.007 min^?1 at [H₂O₂] = 1.0 mM, 27 °C, phosphate buffer 5.0 mM, pH 7.0. Results showed that inactivation and inhibition of microperoxidase as a peroxidase model enzyme occurred simultaneously even at low concentrations of hydrogen peroxide (0.4 mM). This kinetic analysis based on the suicide-substrate inactivation of microperoxidase-11, provides a tool and model for studying peroxidase models in the presence of reversible inhibitors. The introduced inhibition procedure can be used in designing activity tunable and specific protected enzyme models in the hidden and reversibly inhibited forms, which do not undergo inactivation.     

Effects of redox potential control on succinic acid production by engineered Escherichia coli under anaerobic conditions

The effects of oxido-reduction potential (ORP) control on succinic acid production have been investigated in Escherichia coli LL016. In LL016, two CO₂ fixation pathways were achieved and NAD⁺ supply was enhanced by co-expression of heterologous pyruvate carboxylase (PYC) and nicotinic acid phosphoribosyltransferase (NAPRTase). During anaerobic fermentation, cell growth and metabolite distribution were changed with redox potential levels in the range of −200 to −400 mV. From the results, the ORP level of −400 mV was preferable, which resulted in the high succinic acid concentration (28.6 g/L) and high succinic acid productivity (0.33 g/L/h). Meanwhile, the yield of succinic acid at the ORP level of −400 mV was 39% higher than that at the ORP level of −200 mV. In addition, a higher NADH/NAD⁺ ratio and increased enzyme activities were also achieved by regulating the culture to a more reductive environment, which further enhanced the succinic acid production.     

Association between selenium nutritional status and metabolic risk factors in men with visceral obesity

Background and aimPrevious evidence has suggested an association between selenium and cardiovascular disease, which is main outcome of metabolic syndrome. The aim of this study was to examine possible correlation between selenium nutritional status and metabolic risk factors in men with visceral obesity.MethodsPlasma samples were collected from 123 Indonesian men with visceral obesity. Their metabolic risk factors and selenium nutritional status were analyzed. The eligible subjects (n = 78) were stratified according to the International Diabetes Federation: obese, obese plus one component, and obese plus two components or more. Obese plus two components or more were diagnostic criteria of metabolic syndrome. Pearson's correlation was performed to examine the correlation in each group.ResultsIn the obese group, selenium positively correlated with high-density lipoprotein (HDL) cholesterol (r = 0.390, P < 0.05) and with fatty acid binding protein-4 (FABP4) (r = 0.474, P < 0.05); glutathione peroxidase-3 (GPx3) activity was inversely correlated with FABP4 (r = ?467, P < 0.05). In the obese plus one component group, GPx3 activity positively correlated with HDL cholesterol (r = 0.413, P < 0.05). In the metabolic syndrome group, selenium negatively correlated with monocytes chemoattractant protein (MCP)-1 (r = ?0.429, P < 0.05).ConclusionsThese results show that the association between selenium nutritional status and metabolic risk factors is limited to particular group of obese men with or without metabolic syndrome.     

Diatoms as indicators of isolated herbaceous wetland condition in Florida,USA

Benthic, epiphytic, and phytoplanktonic diatoms, as well as soil and water physical–chemical parameters, were sampled from 70 small (average 0.86 ha) isolated depressional herbaceous wetlands located along a gradient of human disturbance in peninsular Florida to (1) compare diatom assemblage structure between algal types; (2) develop biological indicators of wetland condition; (3) examine synecological relationships between diatom structure and environmental variables, with the ultimate goal of developing an index of biological integrity using a single assemblage. Collected diatom samples were enumerated to 250 valves and identified to species or subspecies. An assessment of wetland condition was made using a landscape-scale human disturbance score (Landscape Development Intensity index, LDI), calculated for each site using land use maps and GIS.Assemblages from both impaired and reference sites were compared using blocked multi-response permutation procedures, the percent similarity index, and visually examined using non-metric multidimensional scaling (NMDS). No ecologically significant compositional differences were found within sites. Mantel's test (Mantel's r = 0.29, p < 0.0001) and NMDS (stress: 14.52, variance: 78.5%) identified epiphytic diatoms as the most responsive to human disturbance. Strong significant correlations (|r_s| > 0.50, p < 0.05) were found between epiphytic NMDS site scores and soil pH, specific conductivity, water total phosphorous, and LDI, while soil pH, water color, soil TP, and turbidity were also significantly correlated (p < 0.05).Metrics to assess wetland condition were developed using epiphytic abundance data. Epiphytic taxa sensitive or tolerant to human landscape modification were identified using Indicator Species Analysis, and autecological indices relating diatom sensitivity to nutrients, pH, dissolved oxygen levels, saprobity, salinity, and trophic status were calculated. Fourteen final metrics were identified, scored on an ordinal scale, and combined into the Diatom Index of Wetland Condition (DIWC). The DIWC was highly correlated with the disturbance score (Spearman's r_s = −0.71, p < 0.0001), although the results need to be validated.