Possible Role of Horizontal Gene Transfer in the Colonization of Sea Ice by Algae

Diatoms and other algae not only survive, but thrive in sea ice. Among sea ice diatoms, all species examined so far produce ice-binding proteins (IBPs), whereas no such proteins are found in non-ice-associated diatoms, which strongly suggests that IBPs are essential for survival in ice. The restricted occurrence also raises the question of how the IBP genes were acquired. Proteins with similar sequences and ice-binding activities are produced by ice-associated bacteria, and so it has previously been speculated that the genes were acquired by horizontal transfer (HGT) from bacteria. Here we report several new IBP sequences from three types of ice algae, which together with previously determined sequences reveal a phylogeny that is completely incongruent with algal phylogeny, and that can be most easily explained by HGT. HGT is also supported by the finding that the closest matches to the algal IBP genes are all bacterial genes and that the algal IBP genes lack introns. We also describe a highly freeze-tolerant bacterium from the bottom layer of Antarctic sea ice that produces an IBP with 47% amino acid identity to a diatom IBP from the same layer, demonstrating at least an opportunity for gene transfer. Together, these results suggest that the success of diatoms and other algae in sea ice can be at least partly attributed to their acquisition of prokaryotic IBP genes

Present state and future perspectives of using pluripotent stem cells in toxicology research

The use of novel drugs and chemicals requires reliable data on their potential toxic effects on humans. Current test systems are mainly based on animals or in vitro–cultured animal-derived cells and do not or not sufficiently mirror the situation in humans. Therefore, in vitro models based on human pluripotent stem cells (hPSCs) have become an attractive alternative. The article summarizes the characteristics of pluripotent stem cells, including embryonic carcinoma and embryonic germ cells, and discusses the potential of pluripotent stem cells for safety pharmacology and toxicology. Special attention is directed to the potential application of embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) for the assessment of developmental toxicology as well as cardio- and hepatotoxicology. With respect to embryotoxicology, recent achievements of the embryonic stem cell test (EST) are described and current limitations as well as prospects of embryotoxicity studies using pluripotent stem cells are discussed. Furthermore, recent efforts to establish hPSC-based cell models for testing cardio- and hepatotoxicity are presented. In this context, methods for differentiation and selection of cardiac and hepatic cells from hPSCs are summarized, requirements and implications with respect to the use of these cells in safety pharmacology and toxicology are presented, and future challenges and perspectives of using hPSCs are discussed

Expert consensus document:Cholangiocarcinoma: current knowledge and future perspectives consensus statement from the European Network for the Study of Cholangiocarcinoma (ENS-CCA)

Cholangiocarcinoma (CCA) is a heterogeneous group of malignancies with features of biliary tract differentiation. CCA is the second most common primary liver tumour and the incidence is increasing worldwide. CCA has high mortality owing to its aggressiveness, late diagnosis and refractory nature. In May 2015, the "European Network for the Study of Cholangiocarcinoma" (ENS-CCA: www.enscca.org or www.cholangiocarcinoma.eu) was created to promote and boost international research collaboration on the study of CCA at basic, translational and clinical level. In this Consensus Statement, we aim to provide valuable information on classifications, pathological features, risk factors, cells of origin, genetic and epigenetic modifications and current therapies available for this cancer. Moreover, future directions on basic and clinical investigations and plans for the ENS-CCA are highlighted

In situ expression of eukaryotic ice-binding proteins in microbial communities of Arctic and Antarctic sea ice

Ice-binding proteins (IBPs) have been isolated from various sea-ice organisms. Their characterisation points to a crucial role in protecting the organisms in sub-zero environments. However, their in situ abundance and diversity in natural sea-ice microbial communities is largely unknown. In this study, we analysed the expression and phylogenetic diversity of eukaryotic IBP transcripts from microbial communities of Arctic and Antarctic sea ice. IBP transcripts were found in abundances similar to those of proteins involved in core cellular processes such as photosynthesis. Eighty-nine percent of the IBP transcripts grouped with known IBP sequences from diatoms, haptophytes and crustaceans, but the majority represented novel sequences not previously characterized in cultured organisms. The observed high eukaryotic IBP expression in natural eukaryotic sea ice communities underlines the essential role of IBPs for survival of many microorganisms in communities living under the extreme conditions of polar sea ice

Characterization of Ice-Binding Proteins from Sea Ice Algae

ICE BINDING PROTEINS FROM SEA ICE ALGAE Sea ice is mainly a two-phase system, and its porous structure is largely determinant for biological activity within ice. During ice formation, solutes in the seawater are excluded from the ice matrix and segregate into brine droplets or brine channels, generally defined as brine inclusions inside sea ice. Outflow of high salinity brine and inflow of seawater of lower salinity, as well as further cooling, cause brine inclusions to narrow and eventually separate into individual pockets divided by ice bridges. Despite the harsh conditions that govern within sea ice, where temperatures range from about -1.8°C on the bottom to -20°C or less on the top, and brine salinities can be as high as 200 on the Practical Salinity Scale, brine inclusions offer a habitat for a variety of microalgae. These algae play a crucial role for the ecology of the Polar Oceans, since they represent a concentrated food source in the low-productivity ice-covered sea, and in the months of melting they initiate blooms by seeding the water column. Algae have been found distributed within brine inclusions throughout the entire thickness of the ice column. The strategies adopted by ice microorganisms to cope with conditions in sea ice remain to be unraveled. Recent studies showed that several organisms that populate sea ice, spreading from bacteria to diatoms and a crustacean species, have ice binding proteins (IBPs). These proteins are common in polar species, but lack in temperate organisms, suggesting that IBPs play a key role in adaptation to subzero conditions. The nomenclature of these proteins varies, depending on authors, from ice binding to antifreeze or ice structuring. In the generally accepted adsorption–inhibition model describing the mechanism of action of IBPs, proteins bind to the ice lattice and locally inhibit ice growth by the Gibbs-Thomson effect. Recent publications showed that some IBPs organize water molecules into an ice-like structure that matches defined planes of the ice crystal and is then gradually frozen to the ice lattice. One of the most prominent and best described effects of IBPs is thermal hysteresis, which describes the lowering of the freezing point of a solution below the melting point. Another effect which defines IBPs is inhibition of recrystallization, which is the grain boundary migration resulting in a growth of larger crystals at the expenses of small grains. The biological role of IBPs from sea ice microalgae remains an open question. The importance of some IBP families, as observed in fishes or insects, lies in lowering the freezing point below environmental temperature, in order to avoid ice formation in cells or organs. Other IBPs have the function to inhibit recrystallization, as it has been suggested for plant IBPs. In the context of sea ice, it seems unlikely that the biological role of IBPs may be thermal hysteresis (measured in the order of 1°C) or recrystallization inhibition. Most of the IBPs from sea ice algae are active extracellularly. It has been suggested that they are trapped and accumulate within a layer of extracellular polysaccharide substances (EPS) secreted by several sea ice organisms. Microalgal IBPs produced recombinantly or collected from spent growth medium affect the structure of ice surface, causing pitting and characteristic microstructural features. This suggests that the proteins shape their frozen environment in order to increase their habitable space within sea ice. However, the characterization of IBPs is of relevance not only to understand their functional role in sea ice, but also in the frame of possible applications of IBPs in the medical field, in the food industry and in other fields related to a control of ice crystals. In the following we present some standard techniques to determine the protein activity in terms of thermal hysteresis (TH) and recrystallization inhibition (RI), which define the proteins as ice binding. Also, we present further methods (ice pitting assay, determination of the nucleating temperature) to characterize the activity of IBPs