Chemical defence by sterols in the freshwater ciliate Stentor polymorphus

Heterotrich ciliates typically retain toxic substances in specialized ejectable organelles, called extrusomes, which are used in predator-prey interactions. In this study, we analysed the chemical defence strategy of the freshwater heterotrich ciliate Stentor polymorphus against the predatory ciliate Coleps hirtus, and the microturbellarian flatworm Stenostomum sphagnetorum. The results showed that S. polymorphus is able to defend itself against these two predators by deploying a mix of bioactive sterols contained in its extrusomes. Sterols were isolated in vivo and characterized by liquid chromatography-mass spectrometry (LC-MS), and nuclear magnetic resonance (NMR), as ergosterol, 7-dehydroporiferasterol, and their two peroxidized analogues. The assessment of the toxicity of ergosterol and ergosterol peroxide against various organisms, indicated that these sterols are essential for the effectiveness of the chemical defence in S. polymorphus

lmeEEG: Mass linear mixed-effects modeling of EEG data with crossed random effects

Background: Mixed-effects models are the current standard for the analysis of behavioral studies in psycholinguistics and related fields, given their ability to simultaneously model crossed random effects for subjects and items. However, they are hardly applied in neuroimaging and psychophysiology, where the use of mass univariate analyses in combination with permutation testing would be too computationally demanding to be practicable with mixed models. New method: Here, we propose and validate an analytical strategy that enables the use of linear mixed models (LMM) with crossed random intercepts in mass univariate analyses of EEG data (lmeEEG). It avoids the unfeasible computational costs that would arise from massive permutation testing with LMM using a simple solution: removing random-effects contributions from EEG data and performing mass univariate linear analysis and permutations on the obtained marginal EEG. Results: lmeEEG showed excellent performance properties in terms of power and false positive rate. Comparison with existing methods: lmeEEG overcomes the computational costs of standard available approaches (our method was indeed more than 300 times faster). Conclusions: lmeEEG allows researchers to use mixed models with EEG mass univariate analyses. Thanks to the possibility offered by the method described here, we anticipate that LMM will become increasingly important in neuroscience. Data and codes are available at osf.io/kw87a. The codes and a tutorial are also available at github.com/antovis86/lmeEEG

Evidence for Gene Duplication and Allelic Codominance (not Hierarchical Dominance) at the Mating-Type Locus of the Ciliate, Euplotes crassus.

The high-multiple mating system of Euplotes crassus is known to be controlled by multiple alleles segregating at a single locus and manifesting relationships of hierarchical dominance, so that heterozygous cells would produce a single mating-type substance (pheromone). In strain L-2D, now known to be homozygous at the mating-type locus, we previously identified two pheromones (Ec-α and Ec-1) characterized by significant variations in their amino acid sequences and structure of their macronuclear coding genes. In this study, pheromones and macronuclear coding genes have been analyzed in strain POR-73 characterized by a heterozygous genotype and strong mating compatibility with L-2D strain. It was found that POR-73 cells contain three distinct pheromone coding genes and, accordingly, secrete three distinct pheromones. One pheromone revealed structural identity in amino acid sequence and macronuclear coding gene to the Ec-α pheromone of L-2D cells. The other two pheromones were shown to be new and were designated Ec-2 and Ec-3 to denote their structural homology with the Ec-1 pheromone of L-2D cells. We interpreted these results as evidence of a phenomenon of gene duplication at the E. crassus mating-type locus, and lack of hierarchical dominance in the expression of the macronuclear pheromone genes in cells with heterozygous genotypes

Evolution of mating systems in Euplotes

Ciliates control their sexual phenomenon of conjugation (or mating) through a genetic mechanism of mating types, which may either be only two within a species (recalling the duality of sexes in animals), or multiple (recalling self/non-self compatibility systems in plants and fungi). The nearly one hundred species of the most ubiquitously distributed ciliate, Euplotes, all evolved multiple mating types. Based on analyses of Mendelian genetics, these mating types have for long been assumed to be determined by multi-allelic series of genes inherited at a single genetic locus (i.e., the mating-type or mat locus) and responsible for the synthesis of mating type-pecific signaling proteins. The chemical characterization of these signaling proteins (known as pheromones) from an array of Euplotes species has now permitted us to evolve in the study of Euplotes mating types from an approach of Mendelian genetics to an approach of molecular genetics. In this new experimental context, we have cloned and characterized structurally the pheromone (mating-type) gene families of Euplotes species that take different positions in the phylogenetic tree of the genus Euplotes. It appeared that, in accord with the prediction of the Mendelian genetics, early branching species (e.g., E. polaris, E. raikovi and E. nobilii) inherit their mating types at a single multi-allelic locus. However, in disagreement with the prediction of the Mendelian genetics, late branching species (e.g., E. crassus and E. focardii) inherit their mating types at two distinct loci that are likely the result of an event of gene duplication in the germinal (micronuclear) genome. One locus appears to be structurally and functionally homologous with the multi-allelic locus of the early branching species, while the second locus appears to be structurally homologous but functionally divergent

Francisella-like endosymbionts, potentially harmful to human health, are transported by the universally distributed species of the ciliate Euplotes.

Genome analyses of wild-type strains of two ecologically separated Euplotes species, E. raikovi living in temperate sea waters and E. petzi living in the polar seas, revealed that both host bacteria in their cytoplasm. These bacteria have been identified with facultative intracellular gamma-proteobacteria of the genus Francisella, which includes a number of closely related species well known as extremely infectious to a great variety of organisms. Francisella tularensis, with its four subspecies, is a specialized intracellular pathogen capable of infecting both invertebrate and vertebrate hosts, humans included; F. noatunensis is the etiological agent of the fish disease known as francisellosis, and its two subspecies well adapt to different temperatures of their hosts; the Francisella-like endosymbionts Wolbachia persica, together with the freely living generalists F. philomiragia and F. novicida cause diseases in humans with a compromised immune system. The Francisella endosymbionts of E. raikovi and E. petzi have been successfully isolated and their genomes completely sequenced. They are genetically distant from one another and form two different clades in the Francisella phylogenetic tree, which are distinct from the all other well-established Francisella clades. The finding that Francisella has equally colonized polar and temperate-water species provides evidence that this bacterium is more common and widespread than previously hypothesized, and confirms that free-living Euplotes species and ciliates in general, with their worldwide distribution, may represent a natural reservoir of Francisella in every aquatic environment

Neural Substrates of Mounting Temporal Expectation

A cognitive and neuroanatomical perspective on how timing and expectation are represented in the human brain

Experimental Study of the Effects of Three Types of Meat on Endothelial Function in a Group of Healthy Volunteers

Background: There is a relationship between atherosclerotic risk factors and increased vascular production of reactive oxygen species (ROS). Oxidized LDL and ROS may directly cause endothelial dysfunction by reducing endothelial nitric oxide (NO) bioavailability. The semi-essential amino acid L-arginine is the only substrate for NO synthesis in vascular endothelial cells. Therefore, this amino acid improves endothelial function and plays a role in the prevention and/or treatment of multiple cardiovascular diseases: atherosclerosis, hypertension, diabetes and so on. To determine the effects of three different protein matrices (250 g Fillet of Beef, FB; Chicken Raised on the Ground, CRG; Free-Range Chicken, FRC) with a known content of arginine on the cardiovascular workload, vascular compliance and urinary excretion of some parameters of endothelial function as TGF–Beta, NO (nitrate e nitrite) in a group of healthy volunteers. Materials and methods: We enrolled 10 men to study the behavior of Systolic, Diastolic, Mean, and Pulse Blood Pressure, of Vascular Resistances, of Macro and Micro Vascular Elasticity, of urinary excretion of TGF-β and Nitric Oxide as ratio of creatinine before and after two hours of each meal. The cardiovascular parameters are determined by HDI/Pulse Wave CR 2000 (Hypertension Diagnostic Inc, Eagan, MN); TGF-β is analysed by Elisa method (R&D Systems) and NO by colorimetric method (Cayman). Results and Conclusion: The protein meal packed with CRG causes a significant decrease in diastolic blood pressure mean pressure and vascular resistance in urinary excretion of TGF. FB resulted in a significant decrease in vascular resistance and urinary excretion of NO, while significantly increasing the Pulse Pressure, heart rate and urinary excretion of TGF-β. FRC resulted in a significant reduction of macrovascular elasticity; increase the urinary excretion of TGF and Pulse Pressure. We can conclude that CRG meat looks better both in terms of metabolic and cardiovascular load especially at endothelial level

Ready ... Go: Amplitude of the fMRI Signal Encodes Expectation of Cue Arrival Time

What happens when the brain awaits a signal of uncertain arrival time, as when a sprinter waits for the starting pistol? And what happens just after the starting pistol fires? Using functional magnetic resonance imaging (fMRI), we have discovered a novel correlate of temporal expectations in several brain regions, most prominently in the supplementary motor area (SMA). Contrary to expectations, we found little fMRI activity during the waiting period; however, a large signal appears after the “go” signal, the amplitude of which reflects learned expectations about the distribution of possible waiting times. Specifically, the amplitude of the fMRI signal appears to encode a cumulative conditional probability, also known as the cumulative hazard function. The fMRI signal loses its dependence on waiting time in a “countdown” condition in which the arrival time of the go cue is known in advance, suggesting that the signal encodes temporal probabilities rather than simply elapsed time. The dependence of the signal on temporal expectation is present in “no-go” conditions, demonstrating that the effect is not a consequence of motor output. Finally, the encoding is not dependent on modality, operating in the same manner with auditory or visual signals. This finding extends our understanding of the relationship between temporal expectancy and measurable neural signals