Protocol of single cells preparation for time of flight secondary ion mass spectrometry

There are several techniques like time of flight secondary ion mass spectrometry (ToF SIMS) that require a special protocol for preparation of biological samples, in particular, those containing single cells due to high vacuum conditions that must be kept during the experiment. Frequently, preparation methodology involves liquid nitrogen freezing what is not always convenient. In our studies, we propose and validate a protocol for preparation of single cells. It consists of four steps: (i) paraformaldehyde fixation, (ii) salt removal, (iii) dehydrating, and (iv) sample drying under ambient conditions. The protocol was applied to samples with single melanoma cells i.e. WM115 and WM266-4 characterized by similar morphology. The surface and internal structures of cells were monitored using atomic force, scanning electron and fluorescent microscopes, used to follow any potential protocol-induced alterations. To validate the proposed methodology for sample preparation, ToF SIMS experiments were carried out using C₆₀⁺ cluster ion beam. The applied principal component analysis (PCA) revealed that chemical changes on cell surface of melanoma cells were large enough to differentiate between primary and secondary tumor sites.     

Bone abnormalities in the middle Anisian marine sauropsids from Winterswijk

While the occurrence of skeletal pathologies in Middle Triassic marine reptiles has been poorly documented until now, massive accumulations of bone remains from the Germanic Basin provide the opportunities for documentation. Herein, we describe skeletal abnormalities in the Middle Triassic bone material from the Vossenveld Formation of Winterswijk, the Netherlands. The aim of the study is to distinguish in the studied bones pathologies resulting from malady or trauma and taphonomic alterations. Furthermore, an attempt was made to assess on how the pathologies also represent paleoecological data. Our survey led to the identification of one broken and healed bone, one case of abnormal coossification, and one case of posttraumatic fibro-osseous dysplasia. While the latter two pathologies give little insight into the ecology and function of the affected animals, the fractured dentary is attributed to Nothosaurus marchicus, a common sauropterygian macropredator. It proves that the individual survived long enough to heal, despite the injury hampering its hunting potential. One abnormally shaped humerus is interpreted as postmortem taphonomic deformation, emphasizing the necessity of utilization of detailed diagnostics to distinguish actual paleopathologies from nonbiological distortion.     

Cytonuclear discordance and the species status of Myotis myotis and Myotis blythii (Chiroptera)

We used both highly variable mitochondrial and nuclear loci to investigate the large mouse‐eared bat species complex in the Western Palaearctic to clarify their systematic position. Although mitochondrial lineages show no species segregation and some haplotypes are shared between Myotis myotis and Myotis blythii sensu lato, Bayesian clustering methods based on multilocus genotypes indicate highly concordant nuclear and morphological species assignment. These multilocus, nuclear analyses detected only a single putative F₁ hybrid in the extensive areas of sympatry sampled, thus confirming the biological species status of M. myotis and M. blythii s.l. We propose that the strong cytonuclear discordance in these species complex results from a combination of prior spatial isolation of the two species in different glacial refugia, followed by a succession of mitochondrial introgression events that occurred during the eastward and westward expansions of M. myotis and of M. blythii, respectively. The nuclear markers further indicate the presence of a notable genetic discontinuity within M. myotis that broadly separates populations into an eastern and a western component with an overlap zone in the Balkans. This eastern and western discontinuity is also apparent in the mitochondrial lineages with the D haplogroup largely confined to samples found in Thrace and Asia Minor. None of these genetic discontinuities correspond to the distribution of the two commonly recognized M. myotis subspecies (myotis and macrocephalicus). We also show that distinct morphological subspecies within M. blythii (oxygnathus, omari, risorius and lesviacus) in Europe and the near‐East do not correlate with significant evolutionarily units, whether identified by mitochondrial or nuclear data and thus only represent local morphological variants with little taxonomic relevance.     

An airborne actinobacteria Nocardiopsis alba isolated from bioaerosol of a mushroom compost facility

Actinobacteria are widely distributed in many environments and represent the most important trigger to the occupant respiratory health. Health complaints, including hypersensitivity pneumonitis of the workers, were recorded in a mushroom compost facility (MCF). The studies on the airborne bacteria were carried out to find a possible microbiological source of these symptoms. Culture analysis of compost bioaerosols collected in different location of the MCF was performed. An assessment of the indoor microbial exposure revealed bacterial flora of bioaerosol in the mushroom compost facility represented by Bacillus, Geobacillus, Micrococcus, Pseudomonas, Staphylococcus spp., and actinobacterial strain with white aerial mycelium. The thermotolerant actinobacterial strain of the same morphology was repeatedly isolated from many locations in MCF: air, compost sample, and solid surface in production hall. On the base of complex morphological, chemotaxonomic, and phylogenetic characteristics, the isolate has been classified as Nocardiopsis alba. Dominant position of N. alba in microbial environment of the mushroom compost facility may represent an indicator microorganism in compost bioaerosol. The bioavailability of N. alba in mushroom compost facility creates potential risk for the health of workers, and the protection of respiratory tract and/or skin is strongly recommended.     

Seed maturation and post-harvest ripening negatively affect arabidopsis somatic embryogenesis

Plant Cell, Tissue and Organ Culture (PCTOC) - Plant development is highly malleable, as evidenced by the ability of cultured cells, tissues and organs to regenerate into whole plants in vitro. The...     

Histidine analogues of oxytocin and vasopressin as efficient ligands for Zn ions - Potentiometric and NMR studies

We have characterized the interaction between the Zn²⁺ ions and the histidine analogues of oxytocin and arginine-vasopressin. Potentiometric methods were used for the determination of the stoichiometry of the complexes formed and the calculation of their stability constants. The NMR measurements revealed detailed structures of the complexes and confirmed the binding mode at physiological pH.     

Neonatal asphyxia and hyperthermia and cognitive deficits in adult rats: Role of iron

We compared effects of a critical neonatal anoxia, applied in Wistar rats at body temperatures of 33, 37 and 39 °C, on memory performance in adulthood. Because hyperthermic–anoxic neonates suffer from hyperferremia an additional group of rats, exposed to anoxia at 39 °C, was injected with deferoxamine, a chelator of iron. At the age of 4 and 12 months all rats were examined in hole board, typical maze and Morris maze.     

Uninterrupted growth in a non-polar hadrosaur explains the gigantism among duck-billed dinosaurs

Duck-billed dinosaurs (Hadrosauridae) were the most common ornithopods of the Late Cretaceous. Second only to sauropods and in many cases exceeding the sizes of the largest land mammals (such as indricotheres or proboscideans), they are among the largest terrestrial herbivores to have walked the Earth. Despite their gigantic size, diversity and abundance, their growth strategies remain poorly understood. Here, we examine the bone microstructure of several Mongolian hadrosauroids of varied adult sizes. The small and middle-sized species have lines of arrested growth (LAGs). On the other hand, one of the largest duck-billed dinosaurs, Saurolophus angustirostris, shows uninterrupted growth, comparable with other big hadrosaurs for which the lack of cyclical growth arrests was interpreted as a result of living in the polar region. Since both of the studied taxa inhabited warmer, continental, monsoon-influenced environments of the Late Cretaceous Mongolia, we propose that the absence of LAGs is not a climatic-driven condition but rather connected with the animal's size (i.e. ontogeny). Our results show that, like sauropods, hadrosaurs changed their growth dynamics from cyclical to continuous during their evolution, which made it possible for them to achieve comparable body sizes.