Spontaneous multiscale phase separation within fluorinated xerogel coatings for fouling-release surfaces

Four-component xerogel films consisting of 1 mole-% n-octadecyltrimethoxysilane (C18) and 50 mole-% tetraethoxysilane (TEOS) in combination with 1–24 mole-% tridecafluoro-1,1,2,2-tetrahydrooctyltriethoxysilane (TDF) and 25–48 mole-% n-octyltriethoxysilane (C8) and a 1:49:50 mole-% C18/TDF/TEOS were prepared. Settlement of barnacle cyprids and removal of juvenile barnacles, settlement of zoospores of the alga Ulva linza, and strength of attachment of 7-day sporelings (young plants) of Ulva were compared amongst the xerogel formulations. Several of the xerogel formulations were comparable to poly(dimethylsiloxane) elastomer with respect to removal of juvenile barnacles and removal of sporeling biomass. The 1:4:45:50 and 1:14:35:50 C18/TDF/C8/TEOS xerogels displayed some phase segregation by atomic force microscopy (AFM) pre- and post-immersion in water. Imaging reflectance infrared microscopy showed the formation of islands of alkane-rich and perfluoroalkane-rich regions in these same xerogels both pre- and post-immersion in water. Surface energies were unchanged upon immersion in water for 48 h amongst the TDF-containing xerogel coatings. AFM measurements demonstrated that surface roughness on the 1:4:45:50 and 1:14:35:50 C18/TDF/C8/TEOS xerogel coatings decreased upon immersion in water.     

The control of marine biofouling on xerogel surfaces with nanometer-scale topography

Mixtures of n-octadecyltrimethoxysilane (C18, 1-5 mole-%), n-octyltriethoxysilane (C8) and tetraethoxysilane (TEOS) gave xerogel surfaces of varying topography. The 1:49:50 C18/C8/TEOS xerogel formed 100-400-nm-wide, 2-7-nm deep pores by AFM while coatings with?≥3% C18 were free of such features. Segregation of the coating into alkane-rich and alkane-deficient regions in the 1:49:50 C18/C8/TEOS xerogel was observed by IR microscopy. Immersion in ASW for 48?h gave no statistical difference in surface energy for the 1:49:50 C18/C8/TEOS xerogel and a significant increase for the 50:50 C8/TEOS xerogel. Settlement of barnacle cyprids and removal of juvenile barnacles, settlement of zoospores of the alga Ulva linza, and strength of attachment of 7-day sporelings were compared amongst the xerogel formulations. Settlement of barnacle cyprids was significantly lower in comparison to glass and polystyrene standards. The 1:49:50 and 3:47:50 C18/C8/TEOS xerogels were comparable to PDMSE with respect to removal of juvenile barnacles and sporeling biomass, respectively.     

Spontaneous multiscale phase separation within fluorinated xerogel coatings for fouling-release surfaces

Four-component xerogel films consisting of 1 mole-% n-octadecyltrimethoxysilane (C18) and 50 mole-% tetraethoxysilane (TEOS) in combination with 1-24 mole-% tridecafluoro-1,1,2,2-tetrahydrooctyltriethoxysilane (TDF) and 25-48 mole-% n-octyltriethoxysilane (C8) and a 1:49:50 mole-% C18/TDF/TEOS were prepared. Settlement of barnacle cyprids and removal of juvenile barnacles, settlement of zoospores of the alga Ulva linza, and strength of attachment of 7-day sporelings (young plants) of Ulva were compared amongst the xerogel formulations. Several of the xerogel formulations were comparable to poly(dimethylsiloxane) elastomer with respect to removal of juvenile barnacles and removal of sporeling biomass. The 1:4:45:50 and 1:14:35:50 C18/TDF/C8/TEOS xerogels displayed some phase segregation by atomic force microscopy (AFM) pre- and post-immersion in water. Imaging reflectance infrared microscopy showed the formation of islands of alkane-rich and perfluoroalkane-rich regions in these same xerogels both pre- and post-immersion in water. Surface energies were unchanged upon immersion in water for 48?h amongst the TDF-containing xerogel coatings. AFM measurements demonstrated that surface roughness on the 1:4:45:50 and 1:14:35:50 C18/TDF/C8/TEOS xerogel coatings decreased upon immersion in water.     

The role of surface energy and water wettability in aminoalkyl/fluorocarbon/hydrocarbon-modified xerogel surfaces in the control of marine biofouling

Xerogel films with uniform surface topogrophy, as determined by scanning electron microscopy, atomic force microscopy (AFM), and time-of-flight secondary ion mass spectrometry, were prepared from aminopropylsilyl-, fluorocarbonsilyl-, and hydrocarbonsilyl- containing precursors. Young's modulus was determined from AFM indentation measurements. The xerogel coatings gave reduced settlement of zoospores of the marine fouling alga Ulva compared to a poly(dimethylsiloxane) elastomer (PDMSE) standard. Increased settlement correlated with decreased water wettability as measured by the static water contact angle, θ_Ws, or with decreased polar contribution (γ_P) to the surface free energy (γ_S) as measured by comprehensive contact angle analysis. The strength of attachment of 7-day sporelings (young plants) of Ulva on several of the xerogels was similar to that on PDMSE although no overall correlation was observed with either θ_Ws or γ_S. For sporelings attached to the fluorocarbon/hydrocarbon-modified xerogels, the strength of attachment increased with increased water wettability. The aminopropyl-modified xerogels did not follow this trend.     

Hybrid xerogel films as novel coatings for antifouling and fouling release

Hybrid sol-gel-derived xerogel films prepared from 45/55 (mol ratio) n-propyltrimethoxysilane (C3-TMOS)/tetramethylorthosilane (TMOS), 2/98 (mol ratio) bis[3-(trimethoxysilyl)propyl]-ethylenediamine (enTMOS)/tetraethylorthosilane (TEOS), 50/50 (mol ratio) n-octyltriethoxysilane (C8-TEOS)/TMOS, and 50/50 (mol ratio) 3,3,3-trifluoropropyltrimethoxysilane (TFP-TMOS)/TMOS were found to inhibit settlement of zoospores of the marine fouling alga Ulva (syn. Enteromorpha) relative to settlement on acid-washed glass and give greater release of settled zoospores relative to glass upon exposure to pressure from a water jet. The more hydrophobic 50/50 C8-TEOS/TMOS xerogel films had the lowest critical surface tension by comprehensive contact angle analysis and gave significantly greater release of 8-day Ulva sporeling biomass after exposure to turbulent flow generated by a flow channel than the other xerogel surfaces or glass. The 50/50 C8-TEOS/TMOS xerogel was also a fouling release surface for juveniles of the tropical barnacle Balanus amphitrite. X-ray photon electron data indicated that the alkylsilyl residues of the C3-TMOS-, C8-TEOS-, and TFP-TMOS-containing xerogels were located on the surface of the xerogel films (in a vacuum), which contributes to the film hydrophobicity. Similarly, the amine-containing silyl residues of the enTMOS/TEOS films were located at the surface of the xerogel films, which contributes to the more hydrophilic character and increased critical surface tension of these films.     

Barnacle settlement and the adhesion of protein and diatom microfouling to xerogel films with varying surface energy and water wettability

Previous work has shown that organosilica-based xerogels have the potential to control biofouling. In this study, modifications of chemistry were investigated with respect to their resistance to marine slimes and to settlement of barnacle cyprids. Adhesion force measurements of bovine serum albumin (BSA)-coated atomic force microscopy (AFM) tips to xerogel surfaces prepared from aminopropylsilyl-, fluorocarbonsilyl-, and hydrocarbonsilyl-containing precursors, indicated that adhesion was significantly less on the xerogel surfaces in comparison to a poly(dimethylsiloxane) elastomer (PDMSE) standard. The strength of adhesion of BSA on the xerogels was highest on surfaces with the highest and the lowest critical surface tensions, γ_C and surface energies, γ_S, and duplicated the ‘Baier curve’. The attachment to and removal of cells of the diatom Navicula perminuta from a similar series of xerogel surfaces were examined. Initial attachment of cells was comparable on all of the xerogel surfaces, but the percentage removal of attached cells by hydrodynamic shear stress increased with γ_C and increased wettability as measured by the static water contact angle, θ_Ws, of the xerogel surfaces. The percentage removal of cells of Navicula was linearly correlated with both properties (R ² = 0.74 for percentage removal as a function of θ_Ws and R ² = 0.69 for percentage removal as a function of γ_C). Several of the aminopropylsilyl-containing xerogels showed significantly greater removal of Navicula compared to a PDMSE standard. Cypris larvae of the barnacle B. amphitrite showed preferred settlement on hydrophilic/higher energy surfaces. Settlement was linearly correlated with θ_Ws (R ² = 0.84) and γ_C (R ² = 0.84). Hydrophilic xerogels should prove useful as coatings for boats in regions where fouling is dominated by microfouling (protein and diatom slimes).     

Antifouling character of 'active' hybrid xerogel coatings with sequestered catalysts for the activation of hydrogen peroxide

Halide-permeable xerogel films prepared from sols containing 50 mol% aminopropyltriethoxysilane (APTES)/50 mol% tetraethoxysilane (TEOS) or 10 mol% APTES/90 mol% TEOS and 0.015 M selenoxide or telluride catalyst in the sol gave reduced settlement of cypris larvae of the barnacle Balanus amphitrite and larvae of the tubeworm Hydroides elegans in the presence of artificial seawater (ASW) and hydrogen peroxide (5-100 microM) relative to glass controls. Settlement of Ulva zoospores was lower on both the 50 mol% APTES/50 mol% TEOS and 10 mol% APTES/90 mol% TEOS xerogel formulations in comparison with glass controls with or without the added catalyst. The 50 mol% APTES/50 mol%TEOS xerogel containing telluride catalyst gave reduced settlement of Ulva zoospores in the presence of 100 microM H(2)O(2) in ASW compared with the same coating without added peroxide. Scanning electron microscopy and XPS data suggest that exposure to H(2)O(2) does not lead to chemical or morphological changes on the xerogel surface.     

Correlative assays of barnacle cyprid behaviour for the laboratory evaluation of antifouling coatings: a study of surface energy components

Laboratory evaluation of antifouling coatings is underpinned by settlement studies with specific fouling organisms. Established methods provide insight into the likelihood of failure of a particular coating system, but can neglect the process of surface selection that often precedes attachment. The present approach for quantifying the exploratory behaviour of barnacle cypris larvae suggested that inspection behaviour can be a rapid and predictive proxy for settlement. Two series of xerogels with comparable total surface energy, but different dispersive and polar components, were evaluated. Settlement assays with three-day-old cyprids of Balanus improvisus demonstrated that while attachment was not linked directly to dispersive free energy, the composition of the xerogel was nevertheless significant. Behavioural analysis provided insight into the mechanism of surface rejection. In the case of a 50:50?PH/TEOS (phenyltriethoxysilane-based) xerogel vs a 50:50 TFP/TEOS (3,3,3-trifluoropropyltrimethoxysilane-based) xerogel, wide-searching behaviour was absent on the former.     

The Design of Nanostructured Metronidazole-Loaded HPC/Oxide Xerogel Composites: Influence of the Formulation Parameters on In Vitro Characterisation

In this study, oxide and polymer/oxide xerogels with metronidazole were prepared and examined as carriers of drug for the local application to the bone. The nanoporous SiO₂–CaO–P₂O₅ and HPC–SiO₂–CaO–P₂O₅ xerogel materials with different amounts of the polymer [hydroxypropyl cellulose (HPC)] were prepared using the sol–gel technology, and their physicochemical properties were characterised with respect to chemical structure [by Fourier transform infrared spectroscopy (FTIR)], porosity and the specific surface area of solids (BET), crystallinity [by X-ray powder diffraction (XRD)], morphology [by scanning electron microscope (SEM)] and the in vitro release of the metronidazole over time (by UV–vis spectroscopy, in the ultraviolet light region). HPC-modified oxide xerogels as the carriers of drug showed slower release of metronidazole, due to the structure and stronger interactions with drug as compared with the pure oxide xerogel. Kinetic analysis indicated diffusional mechanism of drug release from all xerogel carriers. HPC addition to the oxide material resulted in a decrease in the porosity and improved the bioactive properties of xerogels. Obtained results for xerogel composites suggest that the metronidazole-loaded xerogels could be attractive candidates for local delivery systems particularly to a bone.KEY WORDS: drug delivery systems, nanostructured composites, porous materials     

干旱胁迫对高山离子芥试管苗抗氧化系统及其活性氧代谢的影响

该试验以高山离子芥试管苗(Chorispora bungeana)为试材,采用固液培养法,设置对照(不添加PEG-6000,CK),轻度干旱胁迫(5%PEG-6000)、中度干旱胁迫(20%PEG-6000)、重度干旱胁迫(40%PEG-6000)4个干旱处理水平,分析干旱胁迫对高山离子芥幼苗抗氧化系统、活性氧代谢等部分生理特征的影响,以揭示高山离子芥在干旱胁迫下的生理响应特征,为进一步探讨其对干旱环境的适应机制奠定基础。结果显示:(1)随着干旱胁迫程度的增加以及在各时间胁迫处理下,抗氧化酶SOD活性及可溶性糖含量显著升高,POD活性、丙二醛含量、CAT活性和APX活性均经历了先升后降的过程。(2)超氧阴离子(O-·2)的产生速率和过氧化氢(H2O2)的含量均显著升高;高山离子芥试管苗叶片相对电导率呈现出升-降-升的变化趋势。(3)相关分析结果显示,MDA与相对电导率、可溶性糖、SOD、APX、O-·2及H2O2呈极显著正相关关系,可溶性糖与SOD、POD、O-·2及H2O2呈极显著正相关关系;相对电导率以及保护酶系均与O-·2、H2O2呈极显著正相关关系。研究表明,高山离子芥具有较强的耐旱性,高山离子芥试管苗在响应干旱胁迫过程中,抗氧化酶系、活性氧代谢、脂质过氧化及渗透调节物等共同参与了高山离子芥试管苗对干旱胁迫的综合抗逆性形成,从而积极启动应对外界干旱环境的耐旱响应机制。     

Enzymatic Saccharification of Pretreated Wheat Straw by T. Reesei Cellulases and A. Niger β-Glucosidase

Three methods of wheat straw treatment (with NaOH, H₂O₂ and butylamine) and its subsequent saccharification by Trichoderma reesei cellulases and Aspergillus niger β-glucosidase are reported. The treatment of straw with NaOH for 1 h in the autoclave (120°C) caused a great loss of the hemicellulose content and a partial removal of lignin, provoking an increase of the cellulose content (from 24% to 69%) in the residue. When the straw was pre-treated with H₂O₂ at 25°C for 20 h, the relative content of cellulose in the straw increased (from 24% to 52%) due to the solubilisation of hemicellulose.

The effect of varying the hydrolysis parameters (enzyme and substrate concentration, temperature and pH) was studied in order to maximise the yield of sugars. Under the best conditions and after 48 h with a mixture of 2:1 (w/w) cellulase/β-glucosidase (with a concentration of 7 and 0.1 U ml^-1, respectively) the native, NaOH-treated and H₂O₂-treated straw material were degraded to reducing sugars for 28%, 89% and 97% respectively.     

Respiration-dependent H2O2 Removal in Brain Mitochondria via the Thioredoxin/Peroxiredoxin System

Mitochondrial reactive oxygen species (ROS) play an important role in both physiological cell signaling processes and numerous pathological states, including neurodegenerative disorders such as Parkinson disease. While mitochondria are considered the major cellular source of ROS, their role in ROS removal remains largely unknown. Using polarographic methods for real-time detection of steady-state H₂O₂ levels, we were able to quantitatively measure the contributions of potential systems toward H₂O₂ removal by brain mitochondria. Isolated rat brain mitochondria showed significant rates of exogenous H₂O₂ removal (9–12 nmol/min/mg of protein) in the presence of substrates, indicating a respiration-dependent process. Glutathione systems showed only minimal contributions: 25% decrease with glutathione reductase inhibition and no effect by glutathione peroxidase inhibition. In contrast, inhibitors of thioredoxin reductase, including auranofin and 1-chloro-2,4-dinitrobenzene, attenuated H₂O₂ removal rates in mitochondria by 80%. Furthermore, a 50% decrease in H₂O₂ removal was observed following oxidation of peroxiredoxin. Differential oxidation of glutathione or thioredoxin proteins by copper (II) or arsenite, respectively, provided further support for the thioredoxin/peroxiredoxin system as the major contributor to mitochondrial H₂O₂ removal. Inhibition of the thioredoxin system exacerbated mitochondrial H₂O₂ production by the redox cycling agent, paraquat. Additionally, decreases in H₂O₂ removal were observed in intact dopaminergic neurons with thioredoxin reductase inhibition, implicating this mechanism in whole cell systems. Therefore, in addition to their recognized role in ROS production, mitochondria also remove ROS. These findings implicate respiration- and thioredoxin-dependent ROS removal as a potentially important mitochondrial function that may contribute to physiological and pathological processes in the brain.     

Changes in respiratory quinone profiles of enhanced biological phosphorus removal activated sludge under different influent phosphorus/carbon ratio conditions

Changes in the microbial community of an enhanced biological phosphorus removal (EBPR) activated sludge system under different influent phosphorus/carbon (P/C) ratio conditions were investigated through evaluation of population respiratory quinone profiles. A total of 13 types of respiratory quinone homologs consisting of 3 types of ubiquinones (UQ) and 10 types of menaquinones (MK) were identified in this study. The dominant quinones were UQ-8 and MK-7 throughout the operational period. A higher P/C ratio (0.1) in the influent stimulated an increase in the mole fractions of UQ-8, MK-7, MK-8(H₄), MK-9(H₄) and MK-8(H₈), suggesting that actinobacterial polyphosphate-accumulating organisms (PAO) containing partially hydrogenated MK, mainly MK-8(H₄), were contributing to EBPR. However, when the P/C ratio gradually decreased from 0.1 to 0.01, the mole fractions of UQ-8 increased from 0.46 to 0.58, while MK-7, MK-8(H₂), MK-8(H₄), MK-9(H₄), MK-8(H₈) and MK-9(H₆) markedly decreased. These changes in the respiratory quinone profiles suggest that glycogen-accumulating organisms corresponding to some Gammaproteobacteria had become dominant populations with a decrease in actinobacterial PAO. On the other hand, increasing abruptly the P/C ratio to 0.1 further caused an increase in the mole fraction of UQ-8, indicating that Rhodocyclus-related organisms were important PAO.     

Influence of chelators and iron ions on the production and degradation of H2O2 by β-amyloid–copper complexes

β-Amyloid peptide (Aβ) 1–42, involved in the pathogenesis of Alzheimer’s disease, binds copper ions to form Aβ · Cu_n complexes that are able to generate H₂O₂ in the presence of a reductant and O₂. The production of H₂O₂ can be stopped with chelators. More reactive than H₂O₂ itself, hydroxyl radicals HO (generated when a reduced redox active metal complex interacts with H₂O₂) are also probably involved in the oxidative stress that creates brain damage during the disease. We report in the present work a method to monitor the effect of chelating agents on the production of hydrogen peroxide by metallo-amyloid peptides. The addition of H₂O₂ associated to a pre-incubation step between ascorbate and Aβ · Cu_n allows to study the formation of H₂O₂ but also, at the same time, its transformation by the copper complexes. Aβ · Cu_n peptides produce but do not efficiently degrade H₂O₂. The reported analytic method, associated to precipitation experiments of copper-containing amyloid peptides, allows to study the inhibition of H₂O₂ production by chelators. The action of a ligand such as EDTA is probably due to the removal of the copper ions from Aβ · Cu_n, whereas bidentate ligands such as 8-hydroxyquinolines probably act via the formation of ternary complexes with Aβ · Cu_n. The redox activity of these bidentate ligands can be modulated by the incorporation or the modification of substituents on the quinoline heterocycle.     

Effects of hydrogen peroxide on relaxation through the NO/sGC/cGMP pathway in isolated rat iliac arteries

Abstract

The production of reactive oxygen species, including hydrogen peroxide (H₂O₂), is increased in diseased blood vessels. Although H₂O₂ leads to impairment of the nitric oxide (NO)/soluble guanylate cyclase (sGC)/cGMP signaling pathway, it is not clear whether this reactive molecule affects the redox state of sGC, a key determinant of NO bioavailability. To clarify this issue, mechanical responses of endothelium-denuded rat external iliac arteries to BAY 41-2272 (sGC stimulator), BAY 60-2770 (sGC activator), nitroglycerin (NO donor), acidified NaNO₂ (exogenous NO) and 8-Br-cGMP (cGMP analog) were studied under exposure to H₂O₂. The relaxant response to BAY 41-2272 (pD₂: 6.79?±?0.10 and 6.62?±?0.17), BAY 60-2770 (pD₂: 9.57?±?0.06 and 9.34?±?0.15) or 8-Br-cGMP (pD₂: 5.19?±?0.06 and 5.24?±?0.08) was not apparently affected by exposure to H₂O₂. In addition, vascular cGMP production stimulated with BAY 41-2272 or BAY 60-2770 in the presence of H₂O₂ was identical to that in its absence. On the other hand, nitroglycerin-induced relaxation was markedly attenuated by exposing the arteries to H₂O₂ (pD₂: 8.73?±?0.05 and 8.30?±?0.05), which was normalized in the presence of catalase (pD₂: 8.59?±?0.05). Likewise, H₂O₂ exposure impaired the relaxant response to acidified NaNO₂ (pD₂: 6.52?±?0.17 and 6.09?±?0.16). These findings suggest that H₂O₂ interferes with the NO-mediated action, but the sGC redox equilibrium and the downstream target(s) of cGMP are unlikely to be affected in the vasculature.     

Inhibition of darkness-induced stomatal closure by ethylene involves a removal of hydrogen peroxide from guard cells of <Emphasis Type="Italic">Vicia faba</Emphasis>

Ethylene regulates many aspects of plant growth and development; however, its effect on the behavior of the stomata is still largely obscure. Here, the association between ethylene inhibition of darkness-induced stomatal closure and hydrogen peroxide (H₂O₂) levels in Vicia faba guard cells was studied. Like ascorbic acid (ASA), the most important reducing substrate for H₂O₂ removal, catalase (CAT), one of H₂O₂-scavenging enzymes, and diphenylene iodonium (DPI), an inhibitor of the H₂O₂-generating enzyme NADPH-oxidase, both ethylene-releasing compound 2-chloroethylene phosphonic acid (ethephon, ETH) and 1-aminocyclopropane-1-carboxylic acid (ACC), the immediate precursor of ethylene, were found to inhibit stomatal closure by darkness and to reduce H₂O₂ levels in guard cells, indicating that ethylene-caused inhibition of darkness-induced stomatal closure involves reduction in the H₂O₂ level in guard cells. Additionally, similar to ASA and CAT, ACC/ETH not only suppressed H₂O₂-induced stomatal closure and H₂O₂ level in guard cells treated with exogenous H₂O₂ in the light, but also reopened the stomata, which had been closed by darkness, and reduced H₂O₂ level that had been generated by darkness, showing that ethylene causes H₂O₂ removal from guard cells. However, the above-mentioned effect of ACC/ETH was dissimilar from that of DPI, which not only was incapable to reduce H₂O₂ level induced by exogenous H₂O₂ but also could not abolish H₂O₂ that had been generated by darkness. Thus, we suggest that ethylene probably induces H₂O₂ removal and reduces H₂O₂ level in guard cells and finally inhibits stomatal closure induced by darkness. Furthermore, the mechanism of H₂O₂ removal caused by ethylene was also discussed.     

Temperature insensitive O2 in blood of the tree frogChiromantis petersi

Summary Respiratory gas exchange and blood respiratory properties have been studied in the East-African tree frogChiromantis petersi. This frog is unusually xerophilous, occupies dry habitats and prefers body temperatures near 40°C and direct solar exposure. Total O₂ uptake was low at 81 l O₂·g^–1·h^–1±19.0 (SD) at 25°C increasing to 253.5 l O₂·g^–1·h^–1±94.8 (SD) at 40°C giving aQ ₁₀ value of 2.1. Skin O₂ uptake at 25°C was 38.5% of total. The gas exchange ratio was 0.71 for whole body gas exchange, 0.61 for the lungs and 1.02 for the skin at 25°C.Blood O₂ affinity was low with aP ₅₀ of 47.5 mmHg at 25°C and pH 7.65. Then _H-value at 25°C increased from 2.7 aroundP ₅₀ to 5.0 at O₂ saturations exceeding 70–80%. Surprisingly, blood O₂ affinity was nearly insensitive to temperature expressed by a H value of ±1.0 kcal·mole between 25 and 40°C.The adaptive significance of the low O₂ affinity, the increase ofn _H with O₂ saturation and the temperature insensitive O₂-Hb binding is discussed in relation to the high and fluctuating body temperatures ofChiromantis.     

Kinetics of hydrogen peroxide elimination by astrocytes and C6 glioma cells: Analysis based on a mathematical model

Oxidative stress is implicated in a variety of disorders including neurodegenerative diseases, and H₂O₂ is important in the generation of reactive oxygen and oxidative stress. In this study, we have examined the rate of extracellular H₂O₂ elimination and relevant enzyme activities in cultured astrocytes and C6 glioma cells and have analyzed the results based on a mathematical model. As compared with other types of cultured cells, astrocytes showed higher activity of glutathione peroxidase (GPx) but lower activities for GSH recycling. C6 cells showed relatively low GPx activity, and treatment of C6 cells with dibutyryl-cAMP, which induces astrocytic differentiation, increased catalase activity and H₂O₂ permeation rate but exerted little effect on other enzyme activities. A mathematical model [N. Makino, K. Sasaki, N. Hashida, Y. Sakakura, A metabolic model describing the H₂O₂ elimination by mammalian cells including H₂O₂ permeation through cytoplasmic and peroxisomal membranes: comparison with experimental data, Biochim. Biophys. Acta 1673 (2004) 149–159.], which includes relevant enzymes and H₂O₂ permeation through membranes, was found to be fitted well to the H₂O₂ concentration dependences of removal reaction with the permeation rate constants as variable parameters. As compared with PC12 cells as a culture model for neuron, H₂O₂ removal activity of astrocytes was considerably higher at physiological H₂O₂ concentrations. The details of the mathematical model are presented in Appendix.     

Evaluation of structural chemistry and isotopic signatures of refractory soil organic carbon fraction isolated by wet oxidation methods

Accurate quantification of different soil organic carbon (SOC) fractions is needed to understand their relative importance in the global C cycle. Among the chemical methods of SOC fractionation, oxidative degradation is considered more promising because of its ability to mimic the natural microbial oxidative processes in soil. This study focuses on detailed understanding of changes in structural chemistry and isotopic signatures of SOC upon different oxidative treatments for assessing the ability of these chemicals to selectively isolate a refractory fraction of SOC. Replicated sampling (to ~1 m depth) of pedons classified as Typic Fragiudalf was conducted under four land uses (woodlot, grassland, no-till and conventional-till continuous corn [Zea mays L.]) at Wooster, OH. Soil samples (<2 mm) were treated with three oxidizing agents (hydrogen peroxide (H₂O₂), disodium peroxodisulfate (Na₂S₂O₈) and sodium hypochlorite (NaOCl)). Oxidation resistant residues and the bulk soil from A1/Ap1 horizons of each land use were further analyzed by solid-state ¹³C nuclear magnetic resonance (NMR) spectroscopy and accelerator mass spectrometry to determine structural chemistry and ¹⁴C activity, respectively. Results indicated that, oxidation with NaOCl removed significantly less SOC compared to Na₂S₂O₈ and H₂O₂. The NMR spectra revealed that NaOCl oxidation preferentially removed lignin-derived compounds at 56 ppm and at 110–160 ppm. On the other hand, the SOC resistant to Na₂S₂O₈ and H₂O₂ oxidation were enriched with alkyl C groups, which dominate in recalcitrant macromolecules. This finding was corroborated by the ¹⁴C activity of residual material, which ranged from ?542 to ?259‰ for Na₂S₂O₈ resistant SOC and ?475 to ?182‰ for H₂O₂ resistant SOC as compared to relatively greater ¹⁴C activity of NaOCl resistant residues (?47 to 61‰). Additionally, H₂O₂ treatment on soils after light fraction removal was more effective in isolating the oldest (¹⁴C activity of ?725 to ?469‰) SOC fraction. The Δ¹⁴C signature of SOC removed by different oxidizing agents, calculated by mass balance, was more or less similar irrespective of the difference in labile SOC removal efficiency. This suggests that SOC isolated by many fractionation methods is still a mixture of much younger and older material and therefore it is very important that the labile SOC should be completely removed before measuring the turnover time of stable and refractory pools of SOC.     

Characterization of hydrogenosomes and their role in glucose metabolism of Neocallimastix sp. L2

In the anaerobic fungus Neocallimastix sp. L2 fermentation of glucose proceeds via the Embden-Meyerhof-Parnas pathway. Enzyme activities leading to the formation of succinate, lactate, ethanol, and formate are associated with the cytoplasmic fraction. The enzymes malic enzyme, NAD(P)H: ferredoxin oxidoreductase, pyruvate: ferredoxin oxidoreductase, hydrogenase, acetate: succinate CoA transferase and succinate thiokinase leading to the formation of H₂, CO₂, acetate, and ATP are localized in microbodies. Thus, these organelles are identified as hydrogenosomes. In addition, the microbodies contain the O₂-scavenging enzymes NADH- and NADPH oxidase, while NAD(P)H peroxidase, catalase, or superoxide dismutase could not be detected. In cell-free extracts from zoospores of Neocallimastix sp. L2 the specific activities of hydrogenosomal enzymes as well as the quantities of these proteins are 2- to 6-fold higher than in mycelium extracts. These findings suggest that hydrogenosomes perform an important role-especially in zoospores — as H₂-evolving, ATP-generating and O₂-scavenging organelles.Abbrevations DTT Dithiotreitol - PEP Phosphoenol pyruvate