Unprecedented within-species chromosome number cline in the Wood White butterfly Leptidea sinapis and its significance for karyotype evolution and speciation

Background: Species generally have a fixed number of chromosomes in the cell nuclei while between-species differences are common and often pronounced. These differences could have evolved through multiple speciation events, each involving the fixation of a single chromosomal rearrangement. Alternatively, marked changes in the karyotype may be the consequence of within-species accumulation of multiple chromosomal fissions/fusions, resulting in highly polymorphic systems with the subsequent extinction of intermediate karyomorphs. Although this mechanism of chromosome number evolution is possible in theory, it has not been well documented. Results: We present the discovery of exceptional intraspecific variability in the karyotype of the widespread Eurasian butterfly Leptidea sinapis. We show that within this species the diploid chromosome number gradually decreases from 2n = 106 in Spain to 2n = 56 in eastern Kazakhstan, resulting in a 6000 km-wide cline that originated recently (8,500 to 31,000 years ago). Remarkably, intrapopulational chromosome number polymorphism exists, the chromosome number range overlaps between some populations separated by hundreds of kilometers, and chromosomal heterozygotes are abundant. We demonstrate that this karyotypic variability is intraspecific because in L. sinapis a broad geographical distribution is coupled with a homogenous morphological and genetic structure. Conclusions: The discovered system represents the first clearly documented case of explosive chromosome number evolution through intraspecific and intrapopulation accumulation of multiple chromosomal changes. Leptidea sinapis may be used as a model system for studying speciation by means of chromosomally-based suppressed recombination mechanisms, as well as clinal speciation, a process that is theoretically possible but difficult to document. The discovered cline seems to represent a narrow time-window of the very first steps of species formation linked to multiple chromosomal changes that have occurred explosively. This case offers a rare opportunity to study this process before drift, dispersal, selection, extinction and speciation erase the traces of microevolutionary events and just leave the final picture of a pronounced interspecific chromosomal difference

Modifications of a method for low energy gamma-ray incident angle reconstruction in the GAMMA-400 gamma-ray telescope

The GAMMA-400 gamma-ray telescope is designed to measure the gamma-ray fluxes in the energy range from 3c20 MeV to 3c1 TeV, performing a sensitive search for high-energy gamma-ray emission when annihilating or decaying dark matter particles. Such measurements will be also associated with the following scientific goals: searching for new and studying known Galactic and extragalactic discrete high-energy gamma-ray sources (supernova remnants, pulsars, accreting objects, microquasars, active galactic nuclei, blazars, quasars). It will be possible to study their structure with high angular resolution and measuring their energy spectra and luminosity with high-energy resolution; identify discrete gamma-ray sources with known sources in other energy ranges. The major advantage of the GAMMA-400 instrument is excellent angular and energy resolutions for gamma rays above 10 GeV. The gamma-ray telescope angular and energy resolutions for the main aperture at 100-GeV gamma rays are 3c0.01% and 3c1%, respectively. The motivation of presented results is to improve physical characteristics of the GAMMA-400 gamma-ray telescope in the energy range of 3c20-100 MeV, most unexplored range today. Such observations are crucial today for a number of high-priority problems faced by modern astrophysics and fundamental physics, including the origin of chemical elements and cosmic rays, the nature of dark matter, and the applicability range of the fundamental laws of physics. To improve the reconstruction accuracy of incident angle for low-energy gamma rays the special analysis of topology of pair-conversion events in thin layers of converter performed. Choosing the pair-conversion events with more precise vertical localization allows us to obtain significantly better angular resolution in comparison with previous and current space and ground-based experiments. For 50-MeV gamma rays the GAMMA-400 gamma-ray telescope angular resolution is better than 50

The work of Josef Váchal in art education at the elementary school

The thesis deals with the art of the modern Czech painter and graphic artist Josef Váchal and follows up its use in creative activities at primary school. The theoretical part is devoted to a detailed analysis of Váchal artistic development and brief psychological study of his personality. It also reflects the creation of other artists, whose artistic expression tends to be close to Váchal works. In fine the thesis follows the possibilities of use of artistic work in art education at the elementary school. The practical part introduce the art education project, inspired by works of Josef Váchal, and documents its complex realization at the elementrary school in Borové Lady with the final reflection

Guidelines for the use and interpretation of assays for monitoring autophagy.

In 2008 we published the first set of guidelines for standardizing research in autophagy. Since then, research on this topic has continued to accelerate, and many new scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Accordingly, it is important to update these guidelines for monitoring autophagy in different organisms. Various reviews have described the range of assays that have been used for this purpose. Nevertheless, there continues to be confusion regarding acceptable methods to measure autophagy, especially in multicellular eukaryotes. A key point that needs to be emphasized is that there is a difference between measurements that monitor the numbers or volume of autophagic elements (e.g., autophagosomes or autolysosomes) at any stage of the autophagic process vs. those that measure flux through the autophagy pathway (i.e., the complete process); thus, a block in macroautophagy that results in autophagosome accumulation needs to be differentiated from stimuli that result in increased autophagic activity, defined as increased autophagy induction coupled with increased delivery to, and degradation within, lysosomes (in most higher eukaryotes and some protists such as Dictyostelium) or the vacuole (in plants and fungi). In other words, it is especially important that investigators new to the field understand that the appearance of more autophagosomes does not necessarily equate with more autophagy. In fact, in many cases, autophagosomes accumulate because of a block in trafficking to lysosomes without a concomitant change in autophagosome biogenesis, whereas an increase in autolysosomes may reflect a reduction in degradative activity. Here, we present a set of guidelines for the selection and interpretation of methods for use by investigators who aim to examine macroautophagy and related processes, as well as for reviewers who need to provide realistic and reasonable critiques of papers that are focused on these processes. These guidelines are not meant to be a formulaic set of rules, because the appropriate assays depend in part on the question being asked and the system being used. In addition, we emphasize that no individual assay is guaranteed to be the most appropriate one in every situation, and we strongly recommend the use of multiple assays to monitor autophagy. In these guidelines, we consider these various methods of assessing autophagy and what information can, or cannot, be obtained from them. Finally, by discussing the merits and limits of particular autophagy assays, we hope to encourage technical innovation in the field