Involvement of an Alternative Oxidase in Oxidative Stress and Mycelium-to-Yeast Differentiation in Paracoccidioides brasiliensis

Paracoccidioides brasiliensis is a thermodimorphic human pathogenic fungus that causes paracoccidioidomycosis (PCM), which is the most prevalent systemic mycosis in Latin America. Differentiation from the mycelial to the yeast form (M-to-Y) is an essential step for the establishment of PCM. We evaluated the involvement of mitochondria and intracellular oxidative stress in M-to-Y differentiation. M-to-Y transition was delayed by the inhibition of mitochondrial complexes III and IV or alternative oxidase (AOX) and was blocked by the association of AOX with complex III or IV inhibitors. The expression of P. brasiliensis aox (Pbaox) was developmentally regulated through M-to-Y differentiation, wherein the highest levels were achieved in the first 24 h and during the yeast exponential growth phase; Pbaox was upregulated by oxidative stress. Pbaox was cloned, and its heterologous expression conferred cyanide-resistant respiration in Saccharomyces cerevisiae and Escherichia coli and reduced oxidative stress in S. cerevisiae cells. These results reinforce the role of PbAOX in intracellular redox balancing and demonstrate its involvement, as well as that of other components of the mitochondrial respiratory chain complexes, in the early stages of the M-to-Y differentiation of P. brasiliensis.

Photoreactions of n-alkyl-3-nitrophenyl ethers with aromatic amines in SDS micelles: A laser flash photolysis study

The quenching of the triplet state of three n-alkyl 3-nitrophenyl ethers: 3-nitroanisol (3-NA), n-butyl 3-nitrophenyl ether (3-NB) and n-decyl 3-nitrophenyl ether (3-ND) by four aniline derivatives: aniline (AN), N,N-dimethylaniline (DMA), 2,4,6-trimethylaniline (TMA), and 4-tetradecylaniline (TDA), was investigated in aqueous micellar SDS solutions by laser flash photolysis. The transient absorption spectra for 3-NA and 3-NB reveal the formation of long-lived intermediate species in the presence of all four quenchers. while for 3-ND no amine-induced intermediates are observed. Comparison of the transient absorption spectra of the probe 3-NA in the presence of DMA in aqueous and micellar solutions shows that the intermediate species are favored by the SDS micelles. With DMA and TMA as quenchers the intermediates are suggested to be the ion radicals generated by single electron transfer from the amine to the probe in the triplet excited state. For the quenchers AN and TDA, the intermediates may be a-complexes. The relative quenching efficiencies generally decrease as the affinity of the quencher for the micellar phase (AN < DMA < TMA < TDA) increases and the mobility of the excited probe (3-NA > 2-NB) decreases. (C) 2011 Elsevier B.V. All rights reserved.

The roles played by Aspergillus nidulans apoptosis-inducing factor (AIF)-like mitochondrial oxidoreductase (AifA) and NADH-ubiquinone oxidoreductases (NdeA-B and NdiA) in farnesol resistance

Farnesol (FOH) is a nonsterol isoprenold produced by dephosphorylanon of farnesyl pyrophosphate a catabolite of the cholesterol biosynthetic pathway These isoprenoids inhibit proliferation and induce apoptosis Here we show that Aspergillus nidulans MA encoding the apoptosis-Inducing factor (AIF)-like mitochondrial oxidoreductase plays a role in the function of the mitochondrial Complex I Additionally we demonstrated that ndeA B and ndiA encode external and internal alternative NADH dehydrogenases respectively that have a function in FOH resistance When exposed to FOH the Delta aifA and Delta ndeA strains have increased ROS production while Delta ndeB Delta ndeA Delta ndeB and Andul mutant strains showed the same ROS accumulation than in the absence of FOH We observed several compensatory mechanisms affecting the differential survival of these mutants to FOH (C) 2010 Elsevier Inc All rights reserved

Investigation of the electrical and electrochemical properties of nanocomposites from V2O5, polypyrrole, and polyaniline

In this work, an investigation of the electrical and electrochemical properties responsible for the energy storage capability of nanocomposites has been carried out. We demonstrate that, in the case of the V2O5 xerogel and the nanocomposites polypyrrole (Ppy)/V2O5 and polyaniline (PANI)/V2O5, the quadratic logistic equation (QLE) can be used to fit the inverse of the resistance values as a function of the injected charge in non-steady-state conditions. This contributes to a phenomenological understanding of the lithium ion and electron transport. The departure of the experimental curve from the fitting observed for the V2O5 xerogel can be attributed to the trapping sites formed during the lithium electroinsertion, which was observed by electrochemical impedance spectroscopy. The amount of trapping sites was obtained on the basis of the QLE. Similar values used to fit the inverse of the resistance were also used to fit the absorbance changes, which is also associated with the small polaron hopping from the V(IV) to the V(V) sites. On the other hand, there was good agreement between the experimental and the theoretical data when the profile of the inverse of the resistance as a function of the amount of inserted lithium ions of the nanocomposites Ppy/V2O5 and PANI/ V2O5 was concerned. We suggest that the presence of the conducting polymers is responsible for the different electrical profile of the V2O5 xerogel compared with those of the nanocomposites. In the latter case, interactions between the lithium ions and oxygen atoms from V2O5 are shielded, thus decreasing the trapping effect of lithium ions in the V2O5 sites. The different values of the lithium ion diffusion coefficient into these intercalation materials are in agreement with this hypothesis.

Effects of N supply on the accumulation of photosynthetic pigments and photoprotectors in Gracilaria tenuistipitata (Rhodophyta) cultured under UV radiation

We have studied the effects of nitrate supply under photosynthetic active radiation (PAR) plus ultraviolet radiation (UVR) exposure on photosynthetic pigments (chlorophyll a and carotenoids), photoprotective UV screen mycosporine-like amino acids (MAAs), and photosynthetic parameters, including the maximum quantum yield (F(v)/F(m)) and electron transport rate (ETR) on the red agarophyte Gracilaria tenuistipitata. Apical tips of G. tenuistipitata were cultivated under ten different concentrations of NO(3)(-) for 7 days. It has been shown that G. tenuistipitata cultured under laboratory conditions has the ability to accumulate high amounts of MAAs following a nitrate concentration-dependent manner under PAR+UVR. Two MAAs were identified, shinorine and porphyra-334. The relative concentration of the first increased under high concentrations of nitrate, while the second one decreased. The presence of antheraxanthin is reported for the first time in this macro-algae, which also contains zeaxanthin, lutein, and beta-carotene. The accumulation of pigments, photoprotective compounds, and photosynthetic parameters of G. tenuistipitata is directly related to N availability. All variables decreased under low N supplies and reached constant maximum values with supplements higher than 0.5 mM NO(3)(-). Our results suggest a high potential to acclimation and photoprotection against stress factors (including high PAR and UVR) directly related to N availability for G. tenuistipitata.

The effects of UV radiation on photosynthesis estimated as chlorophyll fluorescence in Zygnemopsis decussata (Chlorophyta) growing in a high mountain lake (Sierra Nevada, Southern Spain)

The effect of increased UV radiation on photosynthesis estimated as in vivo chlorophyll fluorescence i.e. optimal quantum yield (F(v)/F(m)) and electron transport rate (ETR) in the green filamentous alga Zygnemopsis decussata (Streptophyta, Zygnematales) growing in the high mountain lake ""La Caldera"" (Sierra Nevada, Spain) at 3050 m altitude was evaluated. Two sets of in situ experiments were conducted: (1) On July 2006, F(v)/F(m) was measured throughout the day at different depths (0.1, 0.25, 0.5 and 1 m) and in the afternoon. ETR and phenolic compounds were determined. In addition, in order to analyze the effect of UV radiation, F(v)/F(m) was determined in algae incubated for 3 days at 0.5m under three different light treatments: PAR+UVA+UVB (PAB). PAR+UVA (PA) and PAR (P). (2) On August 2007, F(v)/F(m) was determined under PAB, PA and P treatments and desiccation/rehydration conditions. F(v)/F(m) decreased in algae growing in surface waters (0.1 m) but also at 1 m depth compared to that at 0.5 in depth. The decrease of F(v)/F(m) at noon due to photoinhibition was small (less than 10%) except in algae growing at 1 m depth (44%). The maximal electron transport rate was 3.5-5 times higher in algae growing at 0.25-0.5 m respectively than that at 0.1 and 1 m depth. These results are related to the accumulation of phenolic compounds: i.e. the algae at 0.25-0.5 in presentedrespectively about a 3-5 times higher concentration of phenolic compounds than that of algae at 0.1-1 m depth. The protection mechanisms seem to be stimulated by UVB radiation, since F(v)/F(m) was higher in the presence of UVB (PAB treatment) compared to PA or P treatments. UVA exerts the main photoinhibitory effect, not Only at midday, but also in the afternoon. UVB radiation also had a protective effect in algae grown under desiccation conditions for three days. During re-hydration, the rapid increase of F(v)/F(m) (after 1 h) was higher in the UVB-grown algae than in algae grown under UVA radiation. After 5 h. F(v)/F(m) values were similar in algae submitted to desiccation/rehydration under PAB and P treatments as they were in the control (submerged algae). The combined effect of desiccation and UVA produced the greatest decrease of photosynthesis in Z. decussata. Thifs UVB, in contrast to other species, may support the recovery process. Z. decussata can acclimate to severe stress, conditions in this high mountain lake by the photoprotection mechanism induced by UVB radiation through dynamic photoinhibition and the accumulation of phenolic compounds (UV screen and antioxidant substances).

Molecular Characterization of Propolis-Induced Cell Death in Saccharomyces cerevisiae

Propolis, a natural product of plant resins, is used by the bees to seal holes in their honeycombs and protect the hive entrance. However, propolis has also been used in folk medicine for centuries. Here, we apply the power of Saccharomyces cerevisiae as a model organism for studies of genetics, cell biology, and genomics to determine how propolis affects fungi at the cellular level. Propolis is able to induce an apoptosis cell death response. However, increased exposure to propolis provides a corresponding increase in the necrosis response. We showed that cytochrome c but not endonuclease G (Nuc1p) is involved in propolis-mediated cell death in S. cerevisiae. We also observed that the metacaspase YCA1 gene is important for propolis-mediated cell death. To elucidate the gene functions that may be required for propolis sensitivity in eukaryotes, the full collection of about 4,800 haploid S. cerevisiae deletion strains was screened for propolis sensitivity. We were able to identify 138 deletion strains that have different degrees of propolis sensitivity compared to the corresponding wild-type strains. Systems biology revealed enrichment for genes involved in the mitochondrial electron transport chain, vacuolar acidification, negative regulation of transcription from RNA polymerase II promoter, regulation of macroautophagy associated with protein targeting to vacuoles, and cellular response to starvation. Validation studies indicated that propolis sensitivity is dependent on the mitochondrial function and that vacuolar acidification and autophagy are important for yeast cell death caused by propolis.

Over-expression of COQ10 in Saccharomyces cerevisiae inhibits mitochondrial respiration

COQ10 deletion in Saccharomyces cerevisiae elicits a defect in mitochondrial respiration correctable by addition of coenzyme Q(2). Rescue of respiration by Q(2) is a characteristic of mutants blocked in coenzyme Q(6) synthesis. Unlike Q(6) deficient mutants, mitochondria of the coq10 null mutant have wild-type concentrations Of Q(6). The physiological significance of earlier observations that purified Coq10p contains bound Q(6) was examined in the present study by testing the in vivo effect of over-expression of Coq10p on respiration. Mitochondria with elevated levels of Coq10p display reduced respiration in the bc1 span of the electron transport chain, which can be restored with exogenous Q(2). This suggests that in vivo binding of Q(6) by excess Coq10p reduces the pool of this redox carrier available for its normal function in providing electrons to the bc1 complex. This is confirmed by observing that extra Coq8p relieves the inhibitory effect of excess Coq10p. Coq8p is a putative kinase, and a high-copy suppressor of the coq10 null mutant. As shown here, when over-produced in coq mutants, Coq8p counteracts turnover of Coq3p and Coq4p subunits of the Q-biosynthetic complex. This can account for the observed rescue by COQ8 of the respiratory defect in strains over-producing Coq10p. (C) 2010 Elsevier Inc. All rights reserved.

Intraerythrocytic stages of Plasmodium falciparum biosynthesize menaquinone

Herein, we show that intraerythrocytic stages of Plasmodium falciparum have an active pathway for biosynthesis of menaquinone. Kinetic assays confirmed that plasmodial menaquinone acts at least in the electron transport. Similarly to Escherichia coli, we observed increased levels of menaquinone in parasites kept under anaerobic conditions. Additionally, the mycobacterial inhibitor of menaquinone synthesis Ro 48-8071 also suppressed menaquinone biosynthesis and growth of parasites, although off-targets may play a role in this growth-inhibitory effect. Due to its absence in humans, the menaquinone biosynthesis can be considered an important drug target for malaria. (c) 2010 Federation of European Biochemical Societies. Published by Elsevier B. V. All rights reserved.

Tissue-, substrate-, and site-specific characteristics of mitochondrial reactive oxygen species generation

Reactive oxygen species are a by-product of mitochondrial oxidative phosphorylation, derived from a small quantity of superoxide radicals generated during electron transport. We conducted a comprehensive and quantitative study of oxygen consumption, inner membrane potentials, and H(2)O(2) release in mitochondria isolated from rat brain, heart, kidney, liver, and skeletal muscle, using various respiratory substrates (alpha-ketoglutarate, glutamate, succinate, glycerol phosphate, and palmitoyl carnitine). The locations and properties of reactive oxygen species formation were determined using oxidative phosphorylation and the respiratory chain modulators oligomycin, rotenone, myxothiazol, and antimycin A and the Uncoupler CCCP. We found that in mitochondria isolated from most tissues incubated under physiologically relevant conditions, reactive oxygen release accounts for 0.1-0.2% of O(2) consumed. Our findings support an important participation of flavoenzymes and complex III and a substantial role for reverse electron transport to complex I as reactive oxygen species sources. Our results also indicate that succinate is an important substrate for isolated mitochondrial reactive oxygen production in brain, heart, kidney, and skeletal muscle, whereas fatty acids generate significant quantities of oxidants in kidney and liver. Finally, we found that increasing respiratory rates is an effective way to prevent mitochondrial oxidant release under many, but not all, conditions. Altogether, our data uncover and quantify many tissue-, substrate-, and site-specific characteristics of mitochondrial ROS release. (C) 2009 Elsevier Inc. All rights reserved.

Co-stressors chilling and high light increase photooxidative stress in diuron-treated red alga Kappaphycus alvarezii but with lower involvement of H(2)O(2)

Diuron is one of the most commonly found N-phenylurea herbicides in marine/estuarine waters that promotes toxic effects by inhibiting photosynthesis and affecting the production of reactive oxygen species (ROS) in autotrophs. Since photo- and thermoacclimation are also ROS-mediated processes, this work evaluates a hypothetical additive effect of high light (HL) and chilling (12 degrees C) on 50 nM diuron toxicity to the highly-photosynthetically active apices of the red alga Kappaphycus alvarezii. Additive inhibition of photosynthesis was mainly evidenced by significant decreases of quantum yield of photosystem II and electron transfer rates upon co-stressors exposure to diuron-treated algae. Under extreme 12 degrees C/HL/diuron conditions, unexpected lower correlations between H(2)O(2) concentrations in seawater and radical-sensitive protein thiols were concomitantly measured with the highest indexes of photoinhibition (parameter beta). Altogether, these data support the hypothesis that co-stressors chilling/HL additively inhibit photosynthesis in diuron-exposed K. alvarezii but with less involvement of H(2)O(2) in injury effects than with only chilling or HL. (C) 2010 Elsevier Inc. All rights reserved.

Saccharomyces cerevisiae coq10 null mutants are responsive to antimycin A

Deletion of COQ10 in Saccharomyces cerevisiae elicits a respiratory defect characterized by the absence of cytochrome c reduction, which is correctable by the addition of exogenous diffusible coenzyme Q(2). Unlike other coq mutants with hampered coenzyme Q(6) (Q(6)) synthesis, coq10 mutants have near wild-type concentrations of Q(6). In the present study, we used Q-cycle inhibitors of the coenzyme QH(2)-cytochrome c reductase complex to assess the electron transfer properties of coq10 cells. Our results show that coq10 mutants respond to antimycin A, indicating an active Q-cycle in these mutants, even though they are unable to transport electrons through cytochrome c and are not responsive to myxothiazol. EPR spectroscopic analysis also suggests that wild-type and coq10 mitochondria accumulate similar amounts of Q(6) semiquinone, despite a lower steady-state level of coenzyme QH(2)-cytochrome c reductase complex in the coq10 cells. Confirming the reduced respiratory chain state in coq10 cells, we found that the expression of the Aspergillus fumigatus alternative oxidase in these cells leads to a decrease in antimycin-dependent H(2)O(2) release and improves their respiratory growth.

Transport properties of single vacancies in nanotubes

We present density of states and electronic transport calculations of single vacancies in carbon nanotubes. We confirm that the defect reconstructs into a pentagon and a nonagon, following the removal of a single carbon atom. This leads to the formation of a dangling bond. Finally, we demonstrate that care must be taken when calculating the density of states of impurities in one-dimensional systems in general. Traditional treatments of these systems using periodic boundary conditions leads to the formation of minigaps even in the limit of large unit cells.

Electrical transport in single-crystalline Li(0.9)Mo(6)O(17): A two-band Luttinger liquid exhibiting Bose metal behavior

Temperature-dependent electrical resistance in quasi-one-dimensional Li(0.9)Mo(6)O(17) is described by two Luttinger liquid anomalous exponents alpha, each associated with a distinct one dimensional band. The band with alpha < 1 is argued to crossover to a higher dimension below the temperature T(M'), leading to superconductivity. Disorder and magnetic fields are shown to induce the Bose metal behavior in this bulk compound.

Quantitative constraints on the transport properties of hot partonic matter from semi-inclusive single high transverse momentum pion suppression in Au plus Au collisions at root S(NN)=200 GeV

The PHENIX experiment has measured the suppression of semi-inclusive single high-transverse-momentum pi(0)'s in Au+Au collisions at root s(NN) = 200 GeV. The present understanding of this suppression is in terms of energy loss of the parent (fragmenting) parton in a dense color-charge medium. We have performed a quantitative comparison between various parton energy-loss models and our experimental data. The statistical point-to-point uncorrelated as well as correlated systematic uncertainties are taken into account in the comparison. We detail this methodology and the resulting constraint on the model parameters, such as the initial color-charge density dN(g)/dy, the medium transport coefficient <(q) over cap >, or the initial energy-loss parameter epsilon(0). We find that high-transverse-momentum pi(0) suppression in Au+Au collisions has sufficient precision to constrain these model-dependent parameters at the +/- 20-25% (one standard deviation) level. These constraints include only the experimental uncertainties, and further studies are needed to compute the corresponding theoretical uncertainties.