Distinct properties of semiquinone species detected at the ubiquinol oxidation Q_{o} site of cytochrome bc_{1} and their mechanistic implications

The two-electron ubiquinol oxidation or ubiquinone reduction typically involves semiquinone (SQ) intermediates. Natural engineering of ubiquinone binding sites of bioenergetic enzymes secures that SQ is sufficiently stabilized, so that it does not leave the site to membranous environment before full oxidation/reduction is completed. The ubiquinol oxidation Q(o) site of cytochrome bc(1) (mitochondrial complex III, cytochrome b(6)f in plants) has been considered an exception with catalytic reactions assumed to involve highly unstable SQ or not to involve any SQ intermediate. This view seemed consistent with long-standing difficulty in detecting any reaction intermediates at the Q(o) site. New perspective on this issue is now offered by recent, independent reports on detection of SQ in this site. Each of the described SQs seems to have different spectroscopic properties leaving space for various interpretations and mechanistic considerations. Here, we comparatively reflect on those properties and their consequences on the SQ stabilization, the involvement of SQ in catalytic reactions, including proton transfers, and the reactivity of SQ with oxygen associated with superoxide generation activity of the Q(o) site

Mitochondrial disease-related mutation G167P in cytochrome b of Rhodobacter capsulatus cytochrome bc_{1} (S151P in human) affects the equilibrium distribution of 2Fe-2S cluster and generation of superoxide

Cytochrome bc(1) is one of the key enzymes of many bioenergetic systems. Its operation involves a large scale movement of a head domain of iron-sulfur protein (ISP-HD), which functionally connects the catalytic quinol oxidation Q(o) site in cytochrome b with cytochrome c(1). The Q(o) site under certain conditions can generate reactive oxygen species in the reaction scheme depending on the actual position of ISP-HD in respect to the Q(o) site. Here, using a bacterial system, we show that mutation G167P in cytochrome b shifts the equilibrium distribution of ISP-HD toward positions remote from the Q(o) site. This renders cytochrome bc(1) non-functional in vivo. This effect is remediated by addition of alanine insertions (1Ala and 2Ala) in the neck region of the ISP subunit. These insertions, which on their own shift the equilibrium distribution of ISP-HD in the opposite direction (i.e. toward the Q(o) site), also act in this manner in the presence of G167P. Changes in the equilibrium distribution of ISP-HD in G167P lead to an increased propensity of cytochrome bc(1) to generate superoxide, which becomes evident when the concentration of quinone increases. This result corroborates the recently proposed model in which “semireverse” electron transfer back to the Q(o) site, occurring when ISP-HD is remote from the site, favors reactive oxygen species production. G167P suggests possible molecular effects of S151P (corresponding in sequence to G167P) identified as a mitochondrial disease-related mutation in human cytochrome b. These effects may be valid for other human mutations that change the equilibrium distribution of ISP-HD in a manner similar to G167P

Ectomycorrhizal fungal communities on seedlings and conspecific trees of Pinus mugo grown on the coastal dunes of the Curonian Spit in Lithuania

Ectomycorrhizal (ECM) communities of mature trees and regenerating seedlings of a non-native tree species Pinus mugo grown in a harsh environment of the coastal region of the Curonian Spit National Park in Lithuania were assessed. We established three study sites (S1, S2, and S3) that were separated from each other by 15 km. The ECM species richness was rather low in particular for mature, 100-year-old trees: 12 ectomycorrhizal taxa were identified by molecular analysis from 11 distinguished morphotypes. All 12 taxa were present on seedlings and on mature trees, with between 8–11 and 9–11 taxa present on seedlings and mature trees, respectively. Cenococcum geophilum dominated all ECM communities, but the relative abundance of C. geophilum mycorrhizas was nearly two times higher on seedlings than on mature trees. Mycorrhizal associations formed by Wilcoxina sp., Lactarius rufus, and Russula paludosa were also abundant. Several fungal taxa were only occasionally detected, including Cortinarius sp., Cortinarius obtusus, Cortinarius croceus, and Meliniomyces sp. Shannon’s diversity indices for the ECM assemblages of P. mugo ranged from 0.98 to 1.09 for seedling and from 1.05 to 1.31 for mature trees. According to analysis of similarity, the mycorrhizal communities were similar between the sites (R = 0.085; P = 0.06) and only slightly separated between seedlings and mature trees (R = 0.24; P < 0.0001). An incidental fruiting body survey that was conducted weakly reflected the below-ground assessment of the ECM fungal community and once again showed that ECM and fruiting body studies commonly supply different partial accounts of the true ECM fungal diversity. Our results show that P. mugo has moved into quite distinct habitats and is able to adapt a suite of ECM symbionts that sufficiently support growth and development of this tree and allow for natural seedling regeneration

Diagnosis and management of hyperglycaemia in patients treated with antipsychotic drugs

Research results indicate the presence of an association between mental disorders, certain antipsychotics, and the risk of developing prediabetes (preDM) and specific type diabetes mellitus (DM). However, there are no precise recommendations for their diagnosis and treatment. The obtained data suggest the necessity to perform diagnostics of carbohydrate disorders at the onset of the first symptoms of psychosis, even before the implementation of antipsychotic drugs, and the oral glucose tolerance test (OGTT) seems to be the optimal tool. There is a lot of controversy regarding the timing of control tests addressing the development of dysglycaemia during the use of antipsychotic drugs. We suggest that it should be carried out during the first 4–8 weeks, and in the absence of disorders it should be repeated once a year or with a change in antipsychotic treatment. The diagnostic regimen should then include the need for OGTT supportedby routine determination of the percentage of glycated haemoglobin. If dysglycaemia is diagnosed, the therapeutic management should include non-pharmacological management and hypoglycaemic agents. These recommendations should be individually tailored to each patient and take into account the presence of obesity, which is often found in this group of patients. Weight reduction can be achieved with a properly balanced diet, physical effort, and in justified situations also with drugs effectively reducing body weight. For this reason, drugs are recommended that, if preDM and DM are diagnosed, simultaneously lower glucose levels and reduce body weight. So far, effectiveness in this area has been demonstrated for 2 incretinomimetics: exenatide and liraglutide. Due to the mechanism of preDM/DM development in patients using antipsychotics, the usefulness of other hypoglycaemic drugs with insulin-sensitizing potential — metformin and pioglitazone — has also been suggested. To date, there has been no research on the benefits of other hypoglycaemic drugs in this group of patients

High-resolution cryo-EM structures of plant cytochrome b6fb_{6}f at work

Plants use solar energy to power cellular metabolism. The oxidation of plastoquinol and reduction of plastocyanin by cytochrome $b_{6}f$ (Cyt $b_{6}f$) is known as one of the key steps of photosynthesis, but the catalytic mechanism in the plastoquinone oxidation site ($Q_{p}$) remains elusive. Here, we describe two high-resolution cryo-EM structures of the spinach Cyt $b_{6}f$ homodimer with endogenous plastoquinones and in complex with plastocyanin. Three plastoquinones are visible and line up one after another head to tail near $Q_{p}$ in both monomers, indicating the existence of a channel in each monomer. Therefore, quinones appear to flow through Cyt $b_{6}f$ in one direction, transiently exposing the redox-active ring of quinone during catalysis. Our work proposes an unprecedented one-way traffic model that explains efficient quinol oxidation during photosynthesis and respiration. Structures of cytochrome $b_{6}f$ with and without plastocyanin imply a one-way traffic of quinones for efficient photosynthesis

The coincidence of diabetes mellitus and asthma, their probable causal relationships and therapeutic opportunities

Both the epidemiological data and the everyday medical practice demonstrate the coincidence of various types of diabetes mellitus (DM) in patients with asthma. Specific correlations between the risk of DM in pregnancy, asthma and the consequences of these diseases to the mother and her baby are also explored. The discussion concerning, on the one hand, the impact of asthma-related inflammatory condition on the metabolism of carbohydrates, and, on the other, the presence of chronic hyperglycemia and inflammatory markers observed in patients with asthma, is still ongoing. In the case of asthma and type 1 diabetes mellitus (T1DM), a correlation with the dysfunction of the immune system and the genetic background has been suggested, and in the case of type 2 (T2DM), the vital role of obesity and insulin resistance (IR) to promote excessive proinflammatory immune response. The data indicate that both asthma and DM affect mutually their clinical presentations, including the prognostic values and therapeutic possibilities. The ongoing controversy concerning the effective and safe anti-asthma and hypoglycemizing therapy does not allow for a definitive therapeutic consensus in this group of patients, despite the suggested role of metformin and hyperglycemizing effects of glucocorticoids. Therefore, the objective of the presented paper is a review of the knowledge in the field of DM and asthma coincidence, their probable causal relationships and therapeutic opportunities

How do xanthophylls protect lipid membranes from oxidative damage?

Here, we address the problem of the antioxidant activity of carotenoids in biomembranes. The activity of lutein and zeaxanthin in the quenching of singlet oxygen generated by photosensitization was monitored in lipid vesicles using a singlet oxygen-sensitive fluorescent probe and with the application of fluorescence lifetime imaging microscopy. The antioxidant activity of xanthophylls was interpreted on the basis of electron paramagnetic resonance oximetry results showing that xanthophylls constitute a barrier to the penetration of molecular oxygen into lipid membranes: to a greater extent in the 13-cis configuration than in all-trans. These results are discussed in relation to the trans-cis photoisomerization of xanthophylls observed in the human retina. It can be concluded that photoisomerization of xanthophylls is a regulatory mechanism that is important for both the modulation of light filtration through the macula and photoprotection by quenching singlet oxygen and creating a barrier to oxygen permeation to membranes

Complement inhibitor CSMD1 acts as tumor suppressor in human breast cancer

Human CUB and Sushi multiple domains 1 (CSMD1) is a membrane-bound complement inhibitor suggested to act as a putative tumor suppressor gene, since allelic loss of this region encompassing 8p23 including CSMD1 characterizes various malignancies. Here, we assessed the role of CSMD1 as a tumor suppressor gene in the development of breast cancer in vitro and in vivo. We found that human breast tumor tissues expressed CSMD1 at lower levels compared to that in normal mammary tissues. The decreased expression of CSMD1 was linked to a shorter overall survival of breast cancer patients. We also revealed that expression of CSMD1 in human breast cancer cells BT-20 and MDA-MB-231 significantly inhibited their malignant phenotypes, including migration, adhesion and invasion. Conversely, stable silencing of CSMD1 expression in T47D cells enhanced cancer cell migratory, adherent and clonogenic abilities. Moreover, expression of CSMD1 in the highly invasive MDA-MB-231 cells diminished their signaling potential as well as their stem cell-like properties as assessed by measurement of aldehyde dehydrogenase activity. In a xenograft model, expression of CSMD1 blocked the ability of cancer cells to metastasize to secondary sites in vivo, likely via inhibiting local invasion but not the extravasation into distant tissues. Taken together, these findings demonstrate the role of CSMD1 as a tumor suppressor gene in breast cancer