Heavy fuel oil-contaminated soil remediation by individual and bioaugmentation-assisted phytoremediation with Medicago sativa and with cold plasma-treated M. sativa.

Developing an optimal environmentally friendly bioremediation strategy for petroleum products is of high interest. This study investigated heavy fuel oil (HFO)-contaminated soil (4 and 6 g kg-1) remediation by individual and combined bioaugmentation-assisted phytoremediation with alfalfa (Medicago sativa L.) and with cold plasma (CP)-treated M. sativa. After 14 weeks of remediation, HFO removal efficiency was in the range between 61 and 80% depending on HFO concentration and remediation technique. Natural attenuation had the lowest HFO removal rate. As demonstrated by growth rate and biomass acquisition, M. sativa showed good tolerance to HFO contamination. Cultivation of M. sativa enhanced HFO degradation and soil quality improvement. Bioaugmentation-assisted phytoremediation was up to 18% more efficient in HFO removal through alleviated HFO stress to plants, stimulated plant growth, and biomass acquisition. Cold plasma seed treatment enhanced HFO removal by M. sativa at low HFO contamination and in combination with bioaugmentation it resulted in up to 14% better HFO removal compared to remediation with CP non-treated and non-bioaugmented M. sativa. Our results show that the combination of different remediation techniques is an effective soil rehabilitation strategy to remove HFO and improve soil quality. CP plant seed treatment could be a promising option in soil clean-up and valorization.

Seed Treatment with Electromagnetic Field Induces Different Effects on Emergence, Growth and Profiles of Biochemical Compounds in Seven Half-Sib Families of Silver Birch.

In the context of climate change, strategies aimed at enhancing trees' resistance to biotic and abiotic stress are particularly relevant. We applied an electromagnetic field (EMF) seed treatment to observe changes in the establishment and content of biochemical compounds in silver birch seedlings induced by a short (1 min) seed exposure to a physical stressor. The impact of EMF treatment was evaluated on seedling emergence and growth of one-year-old and two-year-old seedlings from seven half-sib families of silver birch. The effects on numerous biochemical parameters in seedling leaves, such as total phenolic content (TPC), total flavonoid content (TFC), amounts of photosynthetic pigments, total soluble sugars (TSS), level of lipid peroxidation level, antioxidant activity and activity of antioxidant enzymes, were compared using spectrophotometric methods. The results indicated that, in one-year-old seedlings, two of seven (60th and 73rd) half-sib families exhibited a positive response to seed treatment with EMFs in nearly all analyzed parameters. For example, in the 60th family, seed treatment with EMFs increased the percentage of emergence by 3 times, one-year-old seedling height by 71%, leaf TPC by 47%, antioxidant activity by 2 times and amount of chlorophyll a by 4.6 times. Meanwhile, the other two (86th and 179th) families exhibited a more obvious positive response to EMF in two-year-old seedlings as compared to one-year-old seedling controls. The results revealed that short-term EMF treatment of silver birch seeds can potentially be used to improve seedling emergence and growth and increase the content of secondary metabolites, antioxidant capacity and photosynthetic pigments. Understanding of the impact of EMFs as well as the influence of genetic differences on tree responses can be significant for practical applications in forestry. Genetic selection of plant genotypes that exhibit positive response trends can open the way to improve the quality of forest stands.

Comparing Non-Thermal Plasma and Cold Stratification: Which Pre-Sowing Treatment Benefits Wild Plant Emergence?

Meadow restoration and creation projects have faced a lack of local seed diversity due to the limited availability of seed sources. Non-thermal plasma technologies are being developed for agriculture and do not cause damage to heat-sensitive biological systems. This technology has shown the potential to improve agronomic seed quality by enhancing germination and promoting plant growth. However, there is almost no information about the effect of non-thermal plasma pretreatment on the seedlings' emergence of wild plant species. Therefore, this study aimed to evaluate the effect of non-thermal plasma on the emergence of 17 plant seeds originating from local meadows in Lithuania and compare it with the cold stratification pretreatment. The results obtained indicate that there were differences in emergence parameters among the species. However, NTP did not show statistically significant differences from the control. Non-thermal plasma improved the kinetic parameters of emergence for a few specific species' seeds, such as Anthyllis vulneraria and Prunella grandiflora, while the cold stratification pretreatment enhanced emergence for a broader range of plants. Significant differences were observed between non-thermal plasma and stratification pretreatment, as well as between the control and stratification groups. Both methods also had a negative impact.

Cold Plasma-Induced Changes in Stevia rebaudiana Morphometric and Biochemical Parameter Correlations.

Stevia rebaudiana Bertoni is an economically important source of natural low-calorie sweeteners, steviol glycosides (SGs), with stevioside (Stev) and rebaudioside A (RebA) being the most abundant. Pre-sowing seed treatment with cold plasma (CP) was shown to stimulate SGs biosynthesis/accumulation up to several fold. This study aimed to evaluate the possibility to predict CP-induced biochemical changes in plants from morphometric parameters. Principle component analysis (PCA) was applied to two different sets of data: morphometric parameters versus SGs concentrations and ratio, and morphometric parameters versus other secondary metabolites (total phenolic content (TPC), total flavonoid content (TFC)) and antioxidant activity (AA). Seeds were treated for 2, 5 and 7 min with CP (CP2, CP5 and CP7 groups) before sowing. CP treatment stimulated SGs production. CP5 induced the highest increase of RebA, Stev and RebA+Stev concentrations (2.5-, 1.6-, and 1.8-fold, respectively). CP did not affect TPC, TFC or AA and had a duration-dependent tendency to decrease leaf dry mass and plant height. The correlation analysis of individual plant traits revealed that at least one morphometric parameter negatively correlates with Stev orRebA+Stev concentration after CP treatment.

Changes in Content of Bioactive Compounds and Antioxidant Activity Induced in Needles of Different Half-Sib Families of Norway Spruce (Picea abies (L.) H. Karst) by Seed Treatment with Cold Plasma.

In order to ensure sufficient food resources for a constantly growing human population, new technologies (e.g., cold plasma technologies) are being developed for increasing the germination and seedling growth without negative effects on the environment. Pinaceae species are considered a natural source of antioxidant compounds and are valued for their pharmaceutical and nutraceutical properties. In this study, the seeds of seven different Norway spruce half-sib families were processed for one or two minutes with cold plasma (CP) using dielectric barrier discharge (DBD) plasma equipment. At the end of the second vegetation season, the total flavonoid content (TFC), DPPH (2,2- diphenyl-1-picryl-hydrazyl-hydrate), and ABTS (2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid)) antioxidant activity, and the amounts of six organic acids (folic, malic, citric, oxalic, succinic, and ascorbic) were determined in the needles of different half-sib families of Norway spruce seedlings. The results show that the TFC, antioxidant activity, and amounts of organic acids in the seedling needles depended on both the treatment duration and the genetic family. The strongest positive effect on the TFC was determined in the seedlings of the 477, 599, and 541 half-sib families after seed treatment with CP for 1 min (CP1). The TFC in these families increased from 118.06 mg g-1 to 312.6 mg g-1 compared to the control. Moreover, seed treatment with CP1 resulted in the strongest increase in the antioxidant activity of the needles of the 541 half-sib family seedlings; the antioxidant activity, determined by DPPH and ABTS tests, increased by 30 and 23%, respectively, compared to the control. The obtained results indicate that the CP effect on the amount of organic acids in the needles was dependent on the half-sib family. It was determined that treatment with CP1 increased the amount of five organic acids in the needles of the 541 half-sib family seedlings. The presented results show future possibilities for using cold plasma seed treatment in the food and pharmacy industries.

Effects of Non-Thermal Plasma Treatment on Plant Physiological and Biochemical Processes.

Plasma, also called the fourth state of matter, is partially or fully ionized gas [...].

Biochemical and Physiological Plant Processes Affected by Seed Treatment with Non-Thermal Plasma.

Among the innovative technologies being elaborated for sustainable agriculture, one of the most rapidly developing fields relies on the positive effects of non-thermal plasma (NTP) treatment on the agronomic performance of plants. A large number of recent publications have indicated that NTP effects are far more persistent and complex than it was supposed before. Knowledge of the molecular basis and the resulting outcomes of seed treatment with NTP is rapidly accumulating and requires to be analyzed and presented in a systematic way. This review focuses on the biochemical and physiological processes in seeds and plants affected by seed treatment with NTP and the resulting impact on plant metabolism, growth, adaptability and productivity. Wide-scale changes evolving at the epigenomic, transcriptomic, proteomic and metabolic levels are triggered by seed irradiation with NTP and contribute to changes in germination, early seedling growth, phytohormone amounts, metabolic and defense enzyme activity, secondary metabolism, photosynthesis, adaptability to biotic and abiotic stress, microbiome composition, and increased plant fitness, productivity and growth on a longer time scale. This review highlights the importance of these novel findings, as well as unresolved issues that remain to be investigated.

The Potential of Cold Plasma and Electromagnetic Field as Stimulators of Natural Sweeteners Biosynthesis in Stevia rebaudiana Bertoni.

Stevioside (Stev) and rebaudioside A (RebA) are the most abundant steviol glycosides (SGs) responsible for the sweetness of Stevia rabaudiana Bertoni. As compared to Stev, RebA has a higher sweetening potency, better taste and therefore is the most preferred component of the stevia leaf extracts. The aim of this study was to determine the effect of pre-sowing seed treatment with abiotic stressors cold plasma (CP) and electromagnetic field (EMF) on the amount and ratio of RebA and Stev in the leaves of stevia. Additionally, the effect on total phenolic content, flavonoid content and antioxidant activity was investigated. Seeds were treated 5 and 7 min with cold plasma (CP5 and CP7 groups) and 10 min with electromagnetic field (EMF10 group) six days before sowing. The germination tests in vitro demonstrated that all treatments slightly increased germination rate and percentage. HPLC analysis revealed that CP and EMF had strong stimulating effect on SGs accumulation. All treatments increased RebA concentration approximately 1.6-fold; however, the ratio of RebA/Stev decreased from 8.5 in the control to 1.9, 2.5 and 1.1 in CP5, CP7 and EMF10 groups respectively, since the concentration of Stev increased more than RebA, 7.1, 4.6 and 11.0-fold, respectively, compared to control. However, treatments had opposite effect on total phenolic content, flavonoid content, and antioxidant activity. We have demonstrated for the first time that short time pre-sowing treatment of stevia seeds with CP and EMF can be a powerful tool for the enhancement of biosynthesis of RebA and Stev, however it can have negative impact on the content of other secondary metabolites.

Impact of seed color and storage time on the radish seed germination and sprout growth in plasma agriculture.

The use of low-temperature plasma for the pre-sowing seed treatment is still in the early stage of research; thus, numerous factors affecting germination percentage, seedling growth, and yield remains unknown. This study aimed to estimate how two critical factors, such as harvest year and seed coat color, affect the percentage of germination and seedling growth after plasma treatment. Radish seeds stored for 2 and 1 year after harvesting (harvested in 2017 and 2018) were sorted into two colors (brown and grey) to investigate the plasma effect on harvest year and seed coat color. We analyzed the amounts of seed phytohormones and antioxidant (γ-tocopherol) were analyzed using mass spectrometry, and physical changes were studied using SEM, EDX, and EPR to understand the mechanism of plasma-induced changes in radish seeds. The obtained results revealed that plasma treatment on seeds affects the germination kinetics, and the maximal germination percentage depends on seed color and the time of seed storage after harvest. Through this study, for the first time, we demonstrated that physical and chemical changes in radish seeds after plasma treatment depends upon the seed color and harvest year. Positive effects of plasma treatment on growth are stronger for sprouts from seeds harvested in 2017 than in 2018. The plasma treatment effect on the sprouts germinated from grey seeds effect was stronger than sprouts from brown radish seeds. The amounts of gibberellin A3 and abscisic acid in control seeds strongly depended on the seed color, and plasma induced changes were better in grey seeds harvested in 2017. Therefore, this study reveals that Air scalar-DBD plasma's reactive oxygen and nitrogen species (RONS) can efficiently accelerate germination and growth in older seeds.

Cold Plasma Treatment of Sunflower Seeds Modulates Plant-Associated Microbiome and Stimulates Root and Lateral Organ Growth.

Cold atmospheric pressure (CP) plasma irradiation of seeds has been shown to promote plant growth, but the molecular basis of this phenomenon is poorly understood. In our study, optimum irradiation of common sunflower seeds using a dielectric barrier discharge CP device stimulated growth of sunflower lateral organs and roots by 9-14% compared to the control. Metagenomic analysis revealed that the structure of plant-associated bacterial assembly was greatly modified upon CP treatment and could be attributed to the antimicrobial effect of CP-generated reactive species. The treatment resulted in the domination of spore forming Mycobacterium sp. in the above-ground tissues of the seedlings. While the overall bacterial diversity in the roots was barely affected, the CP-induced shift in microbial composition is the likely basis for the observed seedling root growth stimulation and the long-term effect on lateral organ growth and could be mediated by increase in water uptake and/or direct root signaling. Low amplitude protein abundance differences were detected in the roots of the emerging seedlings that are characteristic to low intensity stress stimuli response and could be linked to the changes in plant-associated microbiome upon CP treatment.

Hyperthermia potentiates cisplatin cytotoxicity and negative effects on mitochondrial functions in OVCAR-3 cells.

The aim of this study was to determine the effects of hyperthermia, cisplatin and their combination on mitochondrial functions such as glutamate dehydrogenase (GDH) activity and mitochondrial respiration rates, as well as survival of cultured ovarian adenocarcinoma OVCAR-3 cells. Cells treated for 1 h with hyperthermia (40 and 43 °C) or cisplatin (IC50) or a combination of both treatments were left for recovery at 37 °C temperature for 24 h or 48 h. The obtained results revealed that 43 °C hyperthermia potentiated effects of cisplatin treatment: combinatory treatment more strongly suppressed GDH activity and expression, mitochondrial functions, and decreased survival of OVCAR-3 cells in comparison to separate single treatments. We obtained evidence that in the OVCAR-3 cell line GDH was directly activated by hyperthermia (cisplatin eliminated this effect); however, this effect was followed by GDH inhibition after 48 h recovery. A combination of 43 °C hyperthermia with cisplatin induced stronger GDH inhibition in comparison to separate treatments, and negative effects exerted on GDH activity correlated with suppression of mitochondrial respiration with glutamate + malate. Cisplatin did not induce uncoupling of oxidative phosphorylation in OVCAR-3 cells but induced impairment of the outer mitochondrial membrane in combination with 43 °C hyperthermia. Hyperthermia (43 °C) potentiated cytotoxicity of cisplatin in an OVCAR-3 cell line.

Treatment of Common Sunflower (Helianthus annus L.) Seeds with Radio-frequency Electromagnetic Field and Cold Plasma Induces Changes in Seed Phytohormone Balance, Seedling Development and Leaf Protein Expression.

Treatment of plant seeds with electromagnetic fields or non-thermal plasmas aims to take advantage of plant functional plasticity towards stimulation of plant agricultural performance. In this study, the effects of pre-sowing seed treatment using 200 Pa vacuum (7 min), 5.28 MHz radio-frequency cold plasma (CP -2, 5, and 7 min) and electromagnetic field (EMF -5, 10, 15 min) on seed germination kinetics, content of phytohormones, morphometric parameters of seedlings and leaf proteome were assessed. CP 7 min and EMF 15 min treatments caused 19-24% faster germination in vitro; germination in the substrate was accelerated by vacuum (9%) and EMF 15 min (17%). The stressors did not change the seed germination percentage, with exception of EMF 5 min treatment that caused a decrease by 7.5%. Meanwhile both CP 7 min and EMF 15 min treatments stimulated germination, but the EMF treatment resulted in higher weight of leaves. Stressor-specific changes in phytohormone balance were detected in seeds: vacuum treatment decreased zeatin amount by 39%; CP treatments substantially increased gibberellin content, but other effects strongly varied with the treatment duration; the abscisic acid content was reduced by 55-60% after the EMF treatment. Analysis of the proteome showed that short exposure of seeds to the EMF or CP induced a similar long-term effect on gene expression in leaves, mostly stimulating expression of proteins involved in photosynthetic processes and their regulation.

Impact of Gender and Age on Hyperthermia-Induced Changes in Respiration of Liver Mitochondria.

Aim: This study aimed to compare hyperthermia-induced changes in respiration and generation of reactive oxygen species (ROS) in liver mitochondria derived from animals of different gender and age. Methods: The effects of hyperthermia (40⁻47 °C) on oxidation of different substrates and ROS production were estimated in mitochondria isolated from the liver of male and female rats of the 1⁻1.5, 3⁻4, or 6⁻7 months age. Results: Gender-dependent differences in response of respiration to hyperthermia were the highest at 3⁻4 months of age, less so at 6⁻7 months of age, and only minor at juvenile age. Mild hyperthermia (40⁻42 °C) stimulated pyruvate + malate oxidation in mitochondria of females, but inhibited in mitochondria of males in the 3⁻4 month age group. The resistance of mitochondrial membrane to hyperthermia was the highest at 3⁻4 month males, and the lowest in the 6⁻7 month age group. Inhibition of glutamate + malate oxidation by hyperthermia was caused by thermal inactivation of glutamate dehydrogenase. ROS generation at 37 °C was higher at 1⁻1.5 month of age, but the increase in ROS generation with rise in temperature in this age group was the smallest, and the strongest in 6⁻7 month old animals of both genders. Conclusions: The response to hyperthermia varies during the first 6⁻7 months of life of experimental animals: stronger gender dependence is characteristic at 3⁻4 months of age, while mitochondria from 6⁻7 months animals are less resistant to hyperthermia.

Freshwater bryozoans of Lithuania (Bryozoa).

Nine species of freshwater bryozoans were recorded in Lithuania in a survey of 18 various types of freshwater bodies. Eight species were assigned to the Class Phylactolaemata and families Plumatellidae and Cristatellidae (Plumatella repens, Plumatella fungosa, Plumatella fruticosa, Plumatella casmiana, Plumatella emarginata, Plumatella geimermassardi, Hyalinella punctata and Cristatella mucedo). The ninth species, Paludicella articulata, represented the Class Gymnolaemata. Plumatella geimermassardi and P. casmiana were recorded for the first time in Lithuania. For the plumatellids, species identification was achieved partly by analysing statoblasts' morphological ultrastructures by scanning electron microscopy.

Contribution of mitochondria to injury of hepatocytes and liver tissue by hyperthermia.

OBJECTIVE: The aim of this study was to investigate functional changes of liver mitochondria within the experimentally modeled transition zone of radiofrequency ablation and to estimate possible contribution of these changes to the energy status of liver cells and the whole tissue. MATERIALS AND METHODS: Experiments were carried out on mitochondria isolated from the perfused liver and isolated hepatocytes of male Wistar rats. Hyperthermia was induced by changing the temperature of perfusion medium in the range characteristic for the transition zone (38-52°C). After 15-min perfusion, mitochondria were isolated to investigate changes in the respiration rates and the membrane potential. Adenine nucleotides extracted from isolated hepatocytes and perfused liver subjected to hyperthermic treatment were analyzed by HPLC. RESULTS: Hyperthermic liver perfusion at 42-52°C progressively impaired oxidative phosphorylation in isolated mitochondria. Significant inhibition of the respiratory chain components was observed after perfusion at 42°C, irreversible uncoupling became evident after liver perfusion at higher temperatures (46°C and above). After perfusion at 50-52°C energy supplying function of mitochondria was entirely compromised, and mitochondria turned to energy consumers. Hyperthermia-induced changes in mitochondrial function correlated well with changes in the energy status and viability of isolated hepatocytes, but not with the changes in the energy status of the whole liver tissue. CONCLUSIONS: In this study the pattern of the adverse changes in mitochondrial functions that are progressing with increase in liver perfusion temperature was established. Results of experiments on isolated mitochondria and isolated hepatocytes indicate that hyperthermic treatment significantly and irreversibly inhibits energy-supplying function of mitochondria under conditions similar to those existing in the radiofrequency ablation transition zone and these changes can lead to death of hepatocytes. However, it was not possible to estimate contribution of mitochondrial injury to liver tissue energy status by estimating only hyperthermia-induced changes in adenine nucleotide amounts on the whole tissue level.

Response of perennial woody plants to seed treatment by electromagnetic field and low-temperature plasma.

Radiofrequency (5.28 MHz) electromagnetic radiation and low-temperature plasma were applied as short-term (2-15 min) seed treatments to two perennial woody plant species, including Smirnov's rhododendron (Rhododendron smirnowii Trautv.) and black mulberry (Morus nigra L.). Potential effects were evaluated using germination indices and morphometry. The results suggest that treatment with electromagnetic field stimulated germination of freshly harvested R. smirnowii seeds (increased germination percentage up to 70%), but reduced germination of fresh M. nigra seeds (by 24%). Treatment with low-temperature plasma negatively affected germination for R. smirnowii, and positively for M. nigra. The treatment-induced changes in germination depended on seed dormancy state. Longer-term observations revealed that the effects persisted for more than a year; however, even negative effects on germination came out as positive effects on plant morphometric traits over time. Treatments characterized as distressful based on changes in germination and seedling length increased growth of R. smirnowii after 13 months. Specific changes included stem and root branching, as well as increased leaf count and surface area. These findings imply that longer-term patterns of response to seed stressors may be complex, and therefore, commonly used stressor-effects estimates, such as germination rate or seedling morphology, may be insufficient for qualifying stress response. Bioelectromagnetics. 37:536-548, 2016. © 2016 Wiley Periodicals, Inc.

Hyperthermia differently affects connexin43 expression and gap junction permeability in skeletal myoblasts and HeLa cells.

Stress kinases can be activated by hyperthermia and modify the expression level and properties of membranous and intercellular channels. We examined the role of c-Jun NH2-terminal kinase (JNK) in hyperthermia-induced changes of connexin43 (Cx43) expression and permeability of Cx43 gap junctions (GJs) in the rabbit skeletal myoblasts (SkMs) and Cx43-EGFP transfected HeLa cells. Hyperthermia (42°C for 6 h) enhanced the activity of JNK and its target, the transcription factor c-Jun, in both SkMs and HeLa cells. In SkMs, hyperthermia caused a 3.2-fold increase in the total Cx43 protein level and enhanced the efficacy of GJ intercellular communication (GJIC). In striking contrast, hyperthermia reduced the total amount of Cx43 protein, the number of Cx43 channels in GJ plaques, the density of hemichannels in the cell membranes, and the efficiency of GJIC in HeLa cells. Both in SkMs and HeLa cells, these changes could be prevented by XG-102, a JNK inhibitor. In HeLa cells, the changes in Cx43 expression and GJIC under hyperthermic conditions were accompanied by JNK-dependent disorganization of actin cytoskeleton stress fibers while in SkMs, the actin cytoskeleton remained intact. These findings provide an attractive model to identify the regulatory players within signalosomes, which determine the cell-dependent outcomes of hyperthermia.

Mitochondrial membrane barrier function as a target of hyperthermia.

BACKGROUND AND OBJECTIVE. Hyperthermia is a promising modality for cancer treatment that urgently requires detailed knowledge on molecular and cellular processes for the rational development of treatment protocols. The thorough study of the response of the inner membrane of heart and liver mitochondria to hyperthermia was performed in order to establish the pattern of the hyperthermia-induced changes in the membrane barrier function. MATERIAL AND METHODS. The isolated mitochondria from rat heart and liver (of both genders) were used for experiments, as well as mitochondria isolated from the perfused male rat liver. Changes in the membrane permeability were evaluated by mitochondrial respiration in state 2 or by estimation of the modular kinetics of the membrane leak. RESULTS. The inner membrane of isolated mitochondria from healthy tissues was found to be an extremely sensitive target of hyperthermia that exerted the response even in the febrile range. More severe hyperthermia compromised the inner mitochondrial membrane function; however, this response was tissue-specific and, to some extent, gender-dependent (for liver mitochondria). The data obtained by direct heating of isolated mitochondria were validated by experiments on the perfused liver. CONCLUSIONS. The obtained results imply a crucial importance of the evaluation of the tissue- and gender-specific differences while developing or improving the protocols for hyperthermic treatment or combinatory therapy.

Acute temperature resistance threshold in heart mitochondria: Febrile temperature activates function but exceeding it collapses the membrane barrier.

PURPOSE: Molecular mechanisms underlying hyperthermia-induced cellular injury are not fully understood. The aim of this study was to identify the components of mitochondrial oxidative phosphorylation affected by mild hyperthermia and to quantify the contribution of each component to changes in system behaviour. METHODS: Temperature effects on the oxidative phosphorylation in isolated rat-heart mitochondria were assessed using modular kinetic analysis. Mitochondrial H(2)O(2) production and lipid peroxidation were measured for estimation of temperature-induced oxidative damage. RESULTS: The increase of temperature in the febrile range (40 degrees C) slightly activated mitochondrial function through stimulation of the respiratory module, without affecting the kinetics of the proton leak and phosphorylation modules. At 42 degrees C, state 3 respiration rate remained unchanged, the proton leak across the inner mitochondrial membrane was substantially increased, the respiratory module slightly inhibited, leading to decreased membrane potential (Deltapsi) and diminished ATP synthesis (16% lower phosphorylation flux). Increase of temperature above 42 degrees C caused dissipation of Deltapsi and abolishment of ATP synthesis indicating complete uncoupling of oxidative phosphorylation. The changes in mitochondrial functions induced by incubation at 42 degrees C were completely reversible in contrast to only partial recovery after incubation at higher temperature (45 degrees C). Furthermore, hyperthermia stimulated the production of H(2)O(2) and membrane lipid peroxidation with maximal rates observed at 40 degrees C. CONCLUSIONS: We demonstrated for the first time that febrile temperature (40 degrees C) activates mitochondrial energy supplying functions, whereas further temperature increase by only a few degrees leads to severe impairment of mitochondrial ability to maintain DeltaPsi and synthesise ATP.

Modular kinetic analysis reveals differences in Cd2+ and Cu2+ ion-induced impairment of oxidative phosphorylation in liver.

Impaired mitochondrial function contributes to copper- and cadmium-induced cellular dysfunction. In this study, we used modular kinetic analysis and metabolic control analysis to assess how Cd(2+) and Cu(2+) ions affect the kinetics and control of oxidative phosphorylation in isolated rat liver mitochondria. For the analysis, the system was modularized in two ways: (a) respiratory chain, phosphorylation and proton leak; and (b) coenzyme Q reduction and oxidation, with the membrane potential (Delta psi) and fraction of reduced coenzyme Q as the connecting intermediate, respectively. Modular kinetic analysis results indicate that both Cd(2+) and Cu(2+) ions inhibited the respiratory chain downstream of coenzyme Q. Moreover, Cu(2+), but not Cd(2+) ions stimulated proton leak kinetics at high Delta psi values. Further analysis showed that this difference can be explained by Cu(2+) ion-induced production of reactive oxygen species and membrane lipid peroxidation. In agreement with modular kinetic analysis data, metabolic control analysis showed that Cd(2+) and Cu(2+) ions increased control of the respiratory and phosphorylation flux by the respiratory chain module (mainly because of an increase in the control exerted by cytochrome bc(1) and cytochrome c oxidase), decreased control by the phosphorylation module and increased negative control of the phosphorylation flux by the proton leak module. In summary, we showed that there is a subtle difference in the mode of action of Cd(2+) and Cu(2+) ions on the mitochondrial function, which is related to the ability of Cu(2+) ions to induce reactive oxygen species production and lipid peroxidation.