Regulation of oxidative phosphorylation,the mitochondrial membrane potential,and their role in human disease

Thirty years after Peter Mitchell was awarded the Nobel Prize for the chemiosmotic hypothesis, which links the mitochondrial membrane potential generated by the proton pumps of the electron transport chain to ATP production by ATP synthase, the molecular players involved once again attract attention. This is so because medical research increasingly recognizes mitochondrial dysfunction as a major factor in the pathology of numerous human diseases, including diabetes, cancer, neurodegenerative diseases, and ischemia reperfusion injury. We propose a model linking mitochondrial oxidative phosphorylation (OxPhos) to human disease, through a lack of energy, excessive free radical production, or a combination of both. We discuss the regulation of OxPhos by cell signaling pathways as a main regulatory mechanism in higher organisms, which in turn determines the magnitude of the mitochondrial membrane potential: if too low, ATP production cannot meet demand, and if too high, free radicals are produced. This model is presented in light of the recently emerging understanding of mechanisms that regulate mammalian cytochrome c oxidase and its substrate cytochrome c as representative enzymes for the entire OxPhos system.     

Betacellulin overexpression in the mouse ovary leads to MAPK3/MAPK1 hyperactivation and reduces litter size by impairing fertilization

The epidermal growth factor receptor (EGFR) and its ligands are emerging as key molecules in regulating female reproduction. Here, we used a transgenic mouse model to evaluate whether and at which level of the reproduction cascade higher-than-normal levels of the EGFR ligand betacellulin (BTC) in the reproductive organs affect fertility. Western blots and immunohistochemistry revealed increased BTC levels in uterus and ovaries from transgenic females, particularly evident in granulosa cells of antral follicles. Onset of puberty, estrous cyclicity, and the anatomy and histology of reproductive organs at puberty were not altered as compared to control females. Fertility tests revealed a reduction (~50%) in litter size as the major reproductive deficit of transgenic females. Embryo implantation was delayed in transgenic females, but this was not the reason for the reduced litter size. Transgenic females produced a normal number of oocytes after natural ovulation. The in vivo fertilization rate was significantly reduced in untreated transgenic females but returned to normal levels after superovulation. Impaired oocyte fertilization in the absence of superovulation treatment was associated with MAPK3/MAPK1 hyperactivation in BTC transgenic ovaries, whereas similar levels of MAPK3/MAPK1 activation were detected in transgenic and control ovaries after superovulation treatment. Thus, tight regulation of MAPK3/MAPK1 activity appears to be essential for appropriate granulosa cell function during oocyte maturation. Our study identified hitherto unknown effects of BTC overabundance in reproduction and suggests BTC as a novel candidate protein for the modulation of fertility.     

Ellenberg’s water table experiment put to the test: species optima along a hydrological gradient

An important aspect of niche theory is the position of species’ optima along ecological gradients. It is widely believed that a species’ ecological optimum takes its shape only under competitive pressure. The ecological optimum, therefore, is thought to differ from the physiological optimum in the absence of interspecific competition. Ellenberg’s Hohenheim water table experiment has been very influential in this context. However, the water table gradient in Ellenberg’s experiment was produced by varying the soil thickness above the water table, which confounded the potentially disparate impacts of water table depth (WTD) and soil depth on species growth. Accordingly, here we have re-evaluated Ellenberg’s work. Specifically, we tested the hypothesis that physiological and ecological optima are identical and unaffected by interspecific interaction. We used the same six grasses as in Ellenberg’s experiments, but in our mesocosms, WTD was varied but soil depth kept constant. The design included both an additive component (with/without plant interaction) and a substitutive component (monocultures vs. species mixtures). The results show that the physiological optima along the hydrological gradient varied greatly between species, even in the absence of interspecific interaction. Within species, however, physiological and ecological optima appeared identical in most cases, irrespective of the competition treatment. We conclude that the ‘physiological capacity’ of species largely determines where they are able to persist and that any impact of interspecific interaction is only marginal. These findings are at variance with Ellenberg’s rule, where competition is considered to shift the distribution of a species away from its physiological optimum.     

Hierarchical Unilamellar Vesicles of Controlled Compositional Heterogeneity

Eukaryotic life contains hierarchical vesicular architectures (i.e. organelles) that are crucial for material production and trafficking, information storage and access, as well as energy production. In order to perform specific tasks, these compartments differ among each other in their membrane composition and their internal cargo and also differ from the cell membrane and the cytosol. Man-made structures that reproduce this nested architecture not only offer a deeper understanding of the functionalities and evolution of organelle-bearing eukaryotic life but also allow the engineering of novel biomimetic technologies. Here, we show the newly developed vesicle-in-water-in-oil emulsion transfer preparation technique to result in giant unilamellar vesicles internally compartmentalized by unilamellar vesicles of different membrane composition and internal cargo, i.e. hierarchical unilamellar vesicles of controlled compositional heterogeneity. The compartmentalized giant unilamellar vesicles were subsequently isolated by a separation step exploiting the heterogeneity of the membrane composition and the encapsulated cargo. Due to the controlled, efficient, and technically straightforward character of the new preparation technique, this study allows the hierarchical fabrication of compartmentalized giant unilamellar vesicles of controlled compositional heterogeneity and will ease the development of eukaryotic cell mimics that resemble their natural templates as well as the fabrication of novel multi-agent drug delivery systems for combination therapies and complex artificial microreactors.     

Die einjährigen Arten der GattungArnebia Forsk.

Zusammenfassung Die einjährigen Arten der GattungenArnebia werden, teilsJohnston, teilsPopov folgend, in die zwei UntergattungenArnebia undStrobila, letztere die SektionenStrobila undCornutae umfassend, gegliedert und die insgesamt 16 Arten, von denenA. johnstonii H.Riedl,A. waziristanica H.Riedl undA. simulatrix H.Riedl neu sind, ihrer Morphologie und systematischen Stellung nach besprochen.Durchgeführt mit Unterstützung durch National Science Foundation Grant G 16325.Die Zeichnungen wurden von FrauErika Kubelka ausgeführt, wofür ihr auch an dieser Stelle aufrichtigster Dank gesagt sei.     

A model proposed for the process of evolution with special reference to plants

Summary A spiral running along the surface of a cone standing on top is proposed as a model for evolution, the progress of which is considered as composed of a linear progression following the direction of time and representing the linear increase in the number of taxa, while a circular component stands for the ever recurring functional types of organisms.     

Lack of structure in the gene pool of the highly polyploid ornamental chrysanthemum

Selection of clonally propagated chrysanthemums is mostly performed on F1 hybrids using phenotypic characteristics without the use of molecular information. We applied 448 amplified fragment length polymorphism markers to a set of 81 accessions, mainly from the European gene pool, covering the different horticultural types (cut, pot and garden varieties) and originating from the most important European chrysanthemum breeders. The average pairwise genetic similarity of 0.69 was moderate to rather high. Neighbour-joining clustering resulted in no grouping of the accessions, either by their common origin or their horticultural type, or by similarities in important phenotypic characteristics. The structure of the dendrogram could not be supported by bootstrap analysis. Furthermore, network analysis using SplitsTree, principal coordinate analysis via DARwin or analysis of the population with structure did not differentiate reliable and invariable clusters. Therefore, we tested the marker saturation by plotting the mean coefficient of variation for every pairwise similarity of the bootstrap analysis against the different numbers of markers. We showed that the number of markers is sufficient for a precise estimate of genetic similarity and that the lack of bootstrap support is not due to a low genetic diversity or a lack of marker information, but most likely resulted from the breeding history of the cultivars, involving repeated backcrosses and the exchange of genotypes between breeders.     

Influence of PhoP and Intra-Species Variations on Virulence of Yersinia pseudotuberculosis during the Natural Oral Infection Route

The two-component regulatory system PhoP/PhoQ has been shown to (i) control expression of virulence-associated traits, (ii) confer survival and growth within macrophages and (iii) play a role in Yersinia infections. However, the influence of PhoP on virulence varied greatly between different murine models of infection and its role in natural oral infections with frequently used representative isolates of Y. pseudotuberculosis was unknown. To address this issue, we constructed an isogenic set of phoP ⁺ and phoP ⁻ variants of strain IP32953 and YPIII and analyzed the impact of PhoP using in vitro functionality experiments and a murine oral infection model, whereby we tested for bacterial dissemination and influence on the host immune response. Our results revealed that PhoP has a low impact on virulence, lymphatic and systemic organ colonization, and on immune response modulation by IP32953 and YPIII, indicating that PhoP is not absolutely essential for oral infections but may be involved in fine-tuning the outcome. Our work further revealed certain strain-specific differences in virulence properties, which do not strongly rely on the function of PhoP, but affect tissue colonization, dissemination and/or persistence of the bacteria. Highlighted intra-species variations may provide a potential means to rapidly adjust to environmental changes inside and outside of the host.     

The Human Enzyme That Converts Dietary Provitamin A Carotenoids to Vitamin A Is a Dioxygenase

β-Carotene 15–15′-oxygenase (BCO1) catalyzes the oxidative cleavage of dietary provitamin A carotenoids to retinal (vitamin A aldehyde). Aldehydes readily exchange their carbonyl oxygen with water, making oxygen labeling experiments challenging. BCO1 has been thought to be a monooxygenase, incorporating oxygen from O₂ and H₂O into its cleavage products. This was based on a study that used conditions that favored oxygen exchange with water. We incubated purified recombinant human BCO1 and β-carotene in either ¹⁶O₂-H₂¹⁸O or ¹⁸O₂-H₂¹⁶O medium for 15 min at 37 °C, and the relative amounts of ¹⁸O-retinal and ¹⁶O-retinal were measured by liquid chromatography-tandem mass spectrometry. At least 79% of the retinal produced by the reaction has the same oxygen isotope as the O₂ gas used. Together with the data from ¹⁸O-retinal-H₂¹⁶O and ¹⁶O-retinal-H₂¹⁸O incubations to account for nonenzymatic oxygen exchange, our results show that BCO1 incorporates only oxygen from O₂ into retinal. Thus, BCO1 is a dioxygenase.