Lower Redox State Research Articles

Himalayan Leucogranites: An Experimental Petrology Perspective

The High Himalayan leucogranites (HHL) are produced by muscovite breakdown of a metapelitic source, at temperatures below 800 °C, with initial melt water contents of ~5–7 wt.%. The tourmaline-rich HHL variety is colder, possibly a fractionation product of the hotter two-mica HHL. HHL lack restites such as iron-rich garnet, which, when present, is Mn-rich, signaling fractionation processes. The low redox state of HHL mirrors that of their graphite-bearing source, yet there is evidence of a significant increase in fO2 during crystallization of some HHL. Their relationships with regional deformation call for late emplacement of the main bodies, which must have cooled at 3–4 kb to allow muscovite crystallization, which in turn imposes stringent constraints on unroofing rates of the collisional chain.

Cofactor engineering in Thermoanaerobacterium aotearoense SCUT27 for maximizing ethanol yield and revealing an enzyme complex with high ferredoxin-NAD+ reductase activity

Thermoanaerobacterium aotearoense SCUT27 is a prominent producer of biofuels from lignocellulosic materials. To provide sufficient NAD(P)H for ethanol production, redox-related genes, including lactate dehydrogenase (ldh), redox-sensing transcriptional repressor (rex), and hydrogenase (hfsB), were knocked out. However, the growth of strain PRH (Δldh/Δrex/ΔhfsB) was suppressed due to the intracellular redox state imbalance with the increased NADH concentration. Coincidentally, when the Bcd–EtfAB (BCD) complex was overexpressed, the resulting strain PRH–B3 (Δldh/Δrex/ΔhfsB::BCD) grew rapidly and produced ethanol with a high yield. With lignocellulosic hydrolysates, PRH-BA (Δldh/Δrex/ΔhfsB::BCD::adhE) demonstrated high ethanol productivity and yield, reaching levels of 0.45–0.51 g/L/h and 0.46–0.53 g/g sugars, respectively. The study results shed light on the cofactor balance for cell stability and the high ferredoxin–NAD+ reductase activity of the BCD complex under an intracellular low redox state. They also provide an essential reference for developing strains for improved biofuel production.

Redox potential as a soil health indicator – how does it compare to microbial activity and soil structure?

Background and aimsRedox potential is a promising soil health indicator, which integrates the combined effect of chemical oxidation–reduction reactions into a single measurement. However, this method has been tested only on a few soils. The aim of this study was to test redox potential as a soil health indicator, to see if it responds to management practices and to compare it with existing soil health metrics for microbial activity (“CO2burst”) and soil structure.MethodsWe collected 35 soil samples in agricultural fields from a carbon farming trial, where contrasting management practices have been applied to increase carbon stock. The soil samples were dried, rewetted and analyzed for redox and microbial respiration during rewetting. In addition, soil structure, texture and organic matter content was measured. The data was analyzed for correlations between the indicators and for the differences between management and control fields.ResultsRedox was well correlated with microbial activity, structure, and texture. A low redox state was connected to high microbial activity, indicating bioavailable organic matter availability. Soils with good structure had an oxidized redox status, possibly reflecting high gas-transport. The carbon farming practices resulted in lower oxidation, possibly due to build-up of plant residues.ConclusionsThe findings supported the use of redox as a soil health indicator, but highlighted further research needs for identifying the shared mechanisms linking structure, redox and microbial activity. As such, redox can be a low-cost additional measurement to map changes in soil health, but it cannot replace existing structure or microbial activity measurements.

In Situ Reconstruction of Helical Iron Borophosphate Precatalyst toward Durable Industrial Alkaline Water Electrolysis and Selective Oxidation of Alcohols

AbstractIron‐based (pre)catalysts have attracted enormous attention for various electrooxidation reactions due to the low cost, high abundance, and multiple accessible redox states of iron. Herein, a well‐defined helical iron borophosphate (LiFeBPO) is developed as an electro(pre)catalyst for the oxygen evolution reaction (OER) and selective alcohol oxidation. When deposited on nickel foam (NF), LiFeBPO exhibits an exceptional OER performance at ambient conditions attaining a current density of 100 mA cm−2 at ≈276 mV overpotential in 1 m KOH. Notably, this anode sustains durable alkaline water electrolysis at 500 mA cm−2 for over 330 h under industrial conditions (6 m KOH and 85 °C). In –situ and ex situ investigations reveal a deep reconstruction of LiFeBPO during OER, which transforms into a 3D open porous skeleton assembled by ultrasmall, low‐crystalline α‐FeOOH nanoparticles (interfacing with NiOOH of NF). This structure contributes to exposing accessible surface active sites, as well as accelerating mass transport and bubble detachment. Moreover, this electrode also catalyzes the electrooxidation of alcohols (methanol, ethylene glycol, and glycerol) to formic acid (FA) with high selectivity and full conversion. This study provides promising solutions for designing suitable anodes for the simultaneous production of green hydrogen fuel and value–added FA from electrooxidation reactions.

Human platelet lysate-derived extracellular vesicles enhance angiogenesis through miR-126.

Extracellular vesicles (EVs) are key biological mediators of several physiological functions within the cell microenvironment. Platelets are the most abundant source of EVs in the blood. Similarly, platelet lysate (PL), the best platelet derivative and angiogenic performer for regenerative purposes, is enriched of EVs, but their role is still too poorly discovered to be suitably exploited. Here, we explored the contribution of the EVs in PL, by investigating the angiogenic features extrapolated from that possessed by PL. We tested angiogenic ability and molecular cargo in 3D bioprinted models and by RNA sequencing analysis of PL-derived EVs. A subset of small vesicles is highly represented in PL. The EVs do not retain aggregation ability, preserving a low redox state in human umbilical vein endothelial cells (HUVECs) and increasing the angiogenic tubularly-like structures in 3D endothelial bioprinted constructs. EVs resembled the miRNome profile of PL, mainly enriched with small RNAs and a high amount of miR-126, the most abundant angiogenic miRNA in platelets. The transfer of miR-126 by EVs in HUVEC after the in vitro inhibition of the endogenous form, restored angiogenesis, without involving VEGF as a downstream target in this system. PL is a biological source of available EVs with angiogenic effects involving a miRNAs-based cargo. These properties can be exploited for targeted molecular/biological manipulation of PL, by potentially developing a product exclusively manufactured of EVs.

Evaluation of pollution swapping phenomena from a woodchip denitrification wall targetting removal of nitrate in a shallow gravel aquifer

Woodchip denitrification walls offer a potentially useful way for passive in situ remediation of groundwater nitrate pollution, yet because of the low redox state they induce on the subsurface environment there is an inherent risk they can promote pollution-swapping phenomena. We evaluated pollution-swapping phenomena associated with the first two operational years of a woodchip denitrification wall that is being trialled in a fast-flowing shallow gravel aquifer of quartzo-feldspathic mineralogy. Following burial of woodchip below the water table there was immediate export of dissolved organic carbon (DOC), phosphorus and ammonium into the groundwater. Under the low redox state sustained by labile DOC, the wall initially provided 100% nitrate removal at the expense of acute and localised pollution that occurred in the form of a plume of dissolved iron, manganese and arsenic that were mobilised from the aquifer sediments, in conjunction with methane gas emission. Within one year however, the reactivity of the woodchip wall subsided to support a steady state condition in which nitrate reduction was the terminal electron acceptor process with no measurable methane emission. Having initially functioned as a sink for the potent greenhouse gas nitrous oxide (N2O), evidence is that the woodchip wall is now exporting N2O, albeit at rates less than those associated with productive agricultural land.

Gas release phenomena in soda-lime-silica glass

Heterogeneous bubble nucleation during the production of commercial glasses is one of the negative factors affecting their technology and technical properties. In order to eliminate these bubbles, we conducted extensive research of bubble nucleation in soda-lime-silica glass melts at increasing temperatures and variable redox states of glass. Evolved gas analysis (EGA) coupled with gas chromatography and mass spectrometry were used to identify and evaluate crucial chemical reactions during the melting. The reactions were indicated also by the bubble compositions estimated from bubble contraction during the temperature drop. The fining action of sulfates in the reducing conditions was observed at temperatures between 1200 and 1300 °C using high temperature monitoring. This was confirmed by the relatively high average values of the bubble growth rates. In addition, extensive nucleation of bubbles took place at relatively low temperatures. The temperature region of nucleation was determined by monitoring bubble nucleation on the Pt wire immersed in the melt at increasing temperatures. The nucleation of the bubbles in variously reduced glasses has been observed at temperatures even lower than 1300 °C which further decreased with the increasing C/SO42− ratio. Next, we tried to clarify whether the process of bubble nucleation at a low redox state of glass was caused by supersaturation of the glass melt by either physically dissolved or chemically bound gases. We suppose that the nucleation of bubbles SO2 around 1300 °C can be attributed to the reaction between the sulfate remaining in melts and sulfides which have been formed by reduction reactions.

The viability of primary hepatocytes is maintained under a low cysteine-glutathione redox state with a marked elevation in ophthalmic acid production

Extracellular cystine, the oxidized form of cysteine (Cys), is taken up by cells via the cystine transporter xCT. xCT is not expressed in the liver but is induced in primary hepatocytes under conventional cultured conditions. However, compared to wild-type hepatocytes those from the xCT-knockout mouse showed no evidence of an abnormality and the levels of both Cys and glutathione (GSH) remained unchanged. The levels of ophthalmic acid (OPT), which is produced as an alternative compound by the GSH-synthesizing pathway, became increased during the culturing of hepatocytes. It therefore appears that, in primary hepatocytes, Cys is provided by systems other than xCT, most likely via the transsulfuration pathway, but the levels that are produced are not sufficient. We also employed mouse hepatoma-derived Hepa1-6 cells, which constitutively express xCT. When Hepa 1-6 cells were cultivated in Cys-free media, the levels of intracellular Cys and GSH were decreased, compared to cells cultured in conventional media, leading to cell death accompanied by an increase in the levels of reactive oxygen species and lipid peroxidation products with characteristics similar to ferroptosis. While OPT levels were increased by only to a limited extent in Hepa 1-6 cells, primary hepatocytes cultured in Cys- and Met-free media showed a marked elevation in OPT, reaching levels nearly equivalent to the GSH levels when the cells were cultured in conventional media. Thus, OPT may become a marker for Cys insufficiency and might be used to predict pathological conditions of cells with elevated oxidative stress.

Establishing a Low Redox Potential in Giant Yeast Colonies: Effects of Media and Rotation

Abstract Giant yeast colonies develop a low redox potential, which mimics the electrophilic milieu of both the mitochondrial drug metabolizing compartment and the hypoxic core of many tumors. The major metabolic mediators of low redox potential include: ATP, glutathione, NAD+/NADH, and NADP+/NADPH. Ammonia signaling is the critical mechanism that induces stratification of the giant yeast colonies to allow a low redox potential. A comparison of two powerful investigative models for drug pathways using Saccharomyces cerevisiae have been compounded by the use of different growth media and stimuli to the system. Chemogenetic profiling, which uses a pool of yeast deletion mutants to determine survival changes, is heavily slanted to the use of rich media. Giant yeast colonies studies are heavily slanted to the use of poor media. The current study answers the question “what is the difference over time in redox potential, and its major metabolic mediators, between giant yeast colonies grown on rich and poor media?” Using gene deletion tools, we show that cell death in giant yeast colonies is ammonia-dependent. In poor nutrient, ammonia-depleted (Sok2 deletion mutants) giant yeast cultures, rotation can allow manipulation of reactive oxygen species, providing a model to compare high and low redox states without chemical administration. Mechanistically, these changes are not due to detectable NAD/NAPH or NADP/NADPH changes, but are related in changes in glutathione and ATP concentration.

Validation of Assays for Reactive Oxygen Species and Glutathione in Saccharomyces cerevisiae during Microgravity Simulation

ABSTRACT The effects of spaceflight on yeast have high concordance with agents that induce a very low intracellular redox state and induce a massive efflux of glutathione. These results raise important issues. Can the reduced redox state during spaceflight be reproduced and modulated in ground-based simulations? Will this allow definition of unique drug pathways as a low redox potential state mirrors the electrophilic properties of mitochondria where many drugs are metabolized? Unfortunately, assays for redox status and its major cellular determinant—glutathione—are diverse and often cell-type-specific. Currently, an accepted redox probe set for yeast studies is not available. This paper validates fluorescent probes for glutathione and reactive oxygen status in yeast to support mechanistic studies of microgravity and drug metabolism. The plethora of fluorescent reagents for reactive oxygen species and glutathione makes head-to-head comparisons of all the alternatives impractical. These reagents measure the physiological milieu of reactive oxygen species and diverse thiols, rather than specific individual molecules. We report that in yeast, monochlorobimane (mBCL) and 2’,7’-dichlorodihydrofluorescein diacetate (DC-FDA) are suitable for fluorometric and flow cytometry studies of glutathione and reactive oxygen species, respectively. Both dyes have low background fluorescence, predictable loading, good retention, and are not acutely toxic to Saccharomyces cerevisiae. Both dyes show concordance with other fluorescent and biochemical assays of reactive oxygen species.

Using hollow carbon nanospheres as a light-induced free radical generator to overcome chemotherapy resistance.

Under evolutionary pressure from chemotherapy, cancer cells develop resistance characteristics such as a low redox state, which eventually leads to treatment failures. An attractive option for combatting resistance is producing a high concentration of produced free radicals in situ. Here, we report the production and use of dispersible hollow carbon nanospheres (HCSs) as a novel platform for delivering the drug doxorubicine (DOX) and generating additional cellular reactive oxygen species using near-infrared laser irradiation. These irradiated HCSs catalyzed sufficiently persistent free radicals to produce a large number of heat shock factor-1 protein homotrimers, thereby suppressing the activation and function of resistance-related genes. Laser irradiation also promoted the release of DOX from lysosomal DOX@HCSs into the cytoplasm so that it could enter cell nuclei. As a result, DOX@HCSs reduced the resistance of human breast cancer cells (MCF-7/ADR) to DOX through the synergy among photothermal effects, increased generation of free radicals, and chemotherapy with the aid of laser irradiation. HCSs can provide a unique and versatile platform for combatting chemotherapy-resistant cancer cells. These findings provide new clinical strategies and insights for the treatment of resistant cancers.

Redox chemistry in the phosphorus biogeochemical cycle.

The element phosphorus (P) controls growth in many ecosystems as the limiting nutrient, where it is broadly considered to reside as pentavalent P in phosphate minerals and organic esters. Exceptions to pentavalent P include phosphine--PH3--a trace atmospheric gas, and phosphite and hypophosphite, P anions that have been detected recently in lightning strikes, eutrophic lakes, geothermal springs, and termite hindguts. Reduced oxidation state P compounds include the phosphonates, characterized by C-P bonds, which bear up to 25% of total organic dissolved phosphorus. Reduced P compounds have been considered to be rare; however, the microbial ability to use reduced P compounds as sole P sources is ubiquitous. Here we show that between 10% and 20% of dissolved P bears a redox state of less than +5 in water samples from central Florida, on average, with some samples bearing almost as much reduced P as phosphate. If the quantity of reduced P observed in the water samples from Florida studied here is broadly characteristic of similar environments on the global scale, it accounts well for the concentration of atmospheric phosphine and provides a rationale for the ubiquity of phosphite utilization genes in nature. Phosphine is generated at a quantity consistent with thermodynamic equilibrium established by the disproportionation reaction of reduced P species. Comprising 10-20% of the total dissolved P inventory in Florida environments, reduced P compounds could hence be a critical part of the phosphorus biogeochemical cycle, and in turn may impact global carbon cycling and methanogenesis.

Study of the plutonium (IV) electrochemical behavior in nitric acid at a platinum electrode. Application to the cathodic reduction of Pu(IV) in a plate electrolyzer

Spent fuel reprocessing and notably plutonium recycling contributes in enhancing the sustainability of nuclear energy by preserving uranium natural resources and decreasing the long term radiotoxicity of the final waste. After initial dissolution of the spent nuclear fuel in nitric acid, uranium and plutonium recovery from ultimate waste (fission products and minor actinides) is operated by a liquid–liquid extraction process (PUREX process) and plutonium partitioning from uranium is run on the basis of the selective reduction of Pu and its back extraction. Therefore, the redox properties of Pu in nitric acid are of prime importance and the stabilization of Pu at its lower redox state+III is to be mastered. This study then deals with the characterization of Pu(IV)/Pu(III) redox couple in aqueous nitric acid solution. It was aimed at measuring physico-chemical features of the redox couple such as half-wave and conditional standard potentials, diffusion coefficients and charge transfer rate constant in nitric acid medium. These features have been used in modeling experimental results of preparative Pu(IV) reduction tests in nitrate medium in a laboratory parallel plate electrode cell.

Assessment of macro-micro element accumulation capabilities of elodea nuttallii under gradient redox statuses with elevated nh4-n concentrations

Abstract Aquatic plants often encounter various redox conditions in their natural environment. Elodea nuttallii (Planch.), a submerged aquatic macrophyte, has a flexile ability to use different nutrient sources from various environments. In the present study, Elodea nuttallii was subjected to various redox conditions (+400 mV to -180 mV) at both normal (2.5 ppm) and high (10 ppm) ammonium concentrations and evaluated for macro and micro element accumulation. A reduced environment was prepared by adding glucose to growth medium and nitrogen gas bubbling, while an oxic environment was executed by atmospheric air bubbling. Plants in oxygen-deprived conditions manifested heavy metal (HM) toxicity, such as reduction of biomass and photosynthetic pigments, excess generation of reactive oxygen species (ROS), lipid peroxidation and reduction of major macro elements. In reduced treatments, the bioaccumulation sequence for micro elements was Cu>Mn>Zn>Al>Cd>Fe>Pb at both normal and high NH4-N concentrations. The combined effect of low redox state and high ammonium concentration had a strong physiological impact on the submerged macrophyte. However, macro- and micronutrient accumulation was more significantly affected by reduced environment than by a high NH4-N concentration.

Balancing redox cofactor generation and ATP synthesis: Key microaerobic responses in thermophilic fermentations

Geobacillus thermoglucosidasius is a Gram-positive, thermophilic bacterium capable of ethanologenic fermentation of both C5 and C6 sugars and may have possible use for commercial bioethanol production [Tang et al., 2009; Taylor et al. (2009) Trends Biotechnol 27(7): 398-405]. Little is known about the physiological changes that accompany a switch from aerobic (high redox) to microaerobic/fermentative (low redox) conditions in thermophilic organisms. The changes in the central metabolic pathways in response to a switch in redox potential were analyzed using quantitative real-time PCR and proteomics. During low redox (fermentative) states, results indicated that glycolysis was uniformly up-regulated, the Krebs (tricarboxylic acid or TCA) cycle non-uniformly down-regulated and that there was little to no change in the pentose phosphate pathway. Acetate accumulation was accounted for by strong down-regulation of the acetate CoA ligase gene (acs) in addition to up-regulation of the pta and ackA genes (involved in acetate production), thus conserving ATP while reducing flux through the TCA cycle. Substitution of an NADH dehydrogenase (down-regulated) by an up-regulated NADH:FAD oxidoreductase and up-regulation of an ATP synthase subunit, alongside the observed shifts in the TCA cycle, suggested that an oxygen-scavenging electron transport chain likely remained active during low redox conditions. Together with the observed up-regulation of a glyoxalase and down-regulation of superoxide dismutase, thought to provide protection against the accumulation of toxic phosphorylated glycolytic intermediates and reactive oxygen species, respectively, the changes observed in G. thermoglucosidasius NCIMB 11955 under conditions of aerobic-to-microaerobic switching were consistent with responses to low pO(2) stress.

Agriculture has changed the amount and composition of dissolved organic matter in Central European headwater streams

Dissolved organic matter (DOM) is an important part of the global carbon cycle and significantly influences aquatic ecosystem functions. Recent studies suggest that its amount and composition in freshwaters may be altered by agricultural land use, whereby the influence of preceding in-stream production and processing is not clear. To assess the land use effect on DOM amount and composition for the export from terrestrial to freshwater systems at the land–water interface, we sampled headwater streams draining agricultural and near-pristine catchments (forested and wetland) in the North German plains. To account for spatial and seasonal variation, we conducted a screening of DOM amount (53 sites) and composition (42 sites), and conducted bi-weekly samplings to investigate seasonal variation at eight sites over one year.Concentrations of dissolved organic carbon (DOC) were significantly higher for agricultural and wetland catchments than for forested catchments. Moreover, DOC loads exhibited higher seasonal variation for agricultural and wetland catchments than for forested catchments, which was due to higher variation in discharge. Parallel Factor Analysis revealed that the composition of DOM in agricultural catchments was significantly different from the other studied catchment types, and was characterized by low redox state and high structural complexity. Moreover, a gradient from protein- to humic-like fluorescence significantly separated forested from agricultural and wetland catchments. The contribution of humic-like DOM was strongly and positively related to DOC concentration, suggesting a mechanistic coupling of both. The effects of land use on patterns of DOC concentration and DOM composition were consistent across seasons, implying that land use strongly regulates DOM export. Overall, this study clearly shows the seasonally independent importance of agricultural land use for the amount and composition of DOM fluxes from the terrestrial zone to surface waters. These altered fluxes may affect ecosystem metabolism and health of agricultural headwaters and downstream situated aquatic ecosystems.

Glutathione improves early somatic embryogenesis in Araucaria angustifolia (Bert) O. Kuntze by alteration in nitric oxide emission

In this work, it was observed a straight relationship between the manipulation of the reduced glutathione (GSH)/glutathione disulfide (GSSG) ratio, nitric oxide emission and quality and number of early somatic embryos in Araucaria angustifolia, a Brazilian endangered native conifer. In low concentrations GSH (0.01 and 0.1mM) is a potential NO scavenger in the culture medium. Furthermore, it can increase the number of early SE formed in cell suspension culture media in a few days. However, the maintenance in this low redox state lead to a loss of early somatic embryos polarization. In gelled culture medium, high levels of GSH (5mM) allows the development of globular embryos presenting a high NO emission on embryo apex, stressing its importance in the differentiation and cell division. Taken together these results indicate that the modification of the embryogenic cultures redox state might be an effective strategy to develop more efficient embryogenic systems in A. angustifolia.

Bimodal actions of selenium essential for antioxidant and toxic pro-oxidant activities: The selenium paradox (Review)

Selenium is an essential biological trace element. Adult daily intake of selenium should be approximately 100 µg per day. This compound has a two-sided effect depending on its concentration. A selenium-deficient diet is associated with various endemic diseases, including cardiomuscular malfunctions, osteoarthritis, cancer and viral infections that lead to premature death. These defects are prevented when dietary intake of selenium is adequate. The preventive biological effect of selenium is considered to be due to the antioxidant function of selenoproteins with a selenocysteine in the active site of the catalytic domain. Antioxidant selenoproteins maintain the intracellular redox status and, as a result, normal physiological processes in the cell. Conversely, an overdose of selenium generates oxygen radicals and leads to apoptotic cell death by inducing oxidation and cross-linking of protein thiol groups essential for cell survival. A lower redox state caused by selenium may be implicated in toxic diseases, such as alkali disease and blind staggers. Collectively, selenium seems to have both harmful and beneficial attributes. The aim of this review is to summarize the various biological functions of selenium and to illustrate its opposite roles as a pro-oxidant and an antioxidant.

Murine epidermal progenitor cells have a low redox state

The epidermis is a stratified cornified epithelium that protects the organism against physical and microbial insults. These environmental exposures can result in oxidative stress and affect cellular proliferation and differentiation. However, very few studies have evaluated the base line levels of reactive oxygen species (ROS) and antioxidant proteins in the epidermis or in the context of stem/progenitor cells, cellular types critical to the homeostasis of the epidermis. Here, we show that ROS are mainly produced by epidermal cells and that antioxidant proteins (MnSOD, GPx1, catalase) are differentially expressed in the epidermis. Using our previously sorted epidermal progenitor cells (side population, EpSPs) and more differentiated cells (non‐SPs), we show that catalase expression and activity are significantly decreased in EpSPs compared to non‐SPs. Furthermore, we found fewer peroxisomes per cell and decreased mitochondrial surface area in the EpSPs compared to non‐SPs. Using dihydroethidium (DHE), we show that EpSPs have less DHE signal compared to more differentiated cells. In summary, our data suggest that EpSPs have decreased levels of antioxidant proteins and ROS. Together, these studies indicate that the intracellular redox environment of EpSPs is reduced relative to their more differentiated progeny.Grant Funding SourceNational Institute of Health

Redox state of quinone affects sensitivity of Acanthamoeba castellanii mitochondrial uncoupling protein to purine nucleotides

We studied FFA (free fatty acid)-induced uncoupling activity in Acanthamoeba castellanii mitochondria in the non-phosphorylating state. Either succinate or external NADH was used as a respiratory substrate to determine the proton conductance curves and the relationships between respiratory rate and the quinone reduction level. Our determinations of the membranous quinone reduction level in non-phosphorylating mitochondria show that activation of UCP (uncoupling protein) activity leads to a PN (purine nucleotide)-sensitive decrease in the quinone redox state. The gradual decrease in the rate of quinone-reducing pathways (using titration of dehydrogenase activities) progressively leads to a full inhibitory effect of GDP on LA (linoleic acid) induced proton conductance. This inhibition cannot be attributed to changes in the membrane potential. Indeed, the lack of GDP inhibitory effect observed when the decrease in respiratory rate is accompanied by an increase in the quinone reduction level (using titration of the quinol-oxidizing pathway) proves that the inhibition by nucleotides can be revealed only for a low quinone redox state. It must be underlined that, in A. castellanii non-phosphorylating mitochondria, the transition of the inhibitory effect of GDP on LA-induced UCP-mediated uncoupling is observed for the same range of quinone reduction levels (between 50% and 40%) as that observed previously for phosphorylating conditions. This observation, drawn from the two different metabolic states of mitochondria, indicates that quinone could affect UCP activity through sensitivity to PNs.