Easy and cost-effective stable positioning of suspension cells during live-cell imaging

Dynamic processes of cells can be best monitored when living cells are analyzed by imaging. While it is easy to observe adherent living cells it has been extremely challenging to analyze suspension cells. This cell type floats freely in the culture dish, and it is only a question of time when the focus or the observation field is lost. In order to keep the cells in focus, an easy and inexpensive method allowing the observation of living suspension cells during confocal laser scanning microscope imaging was developed

Identification of Differentially Expressed Genes during Bacillus subtilis Spore Outgrowth in High-Salinity Environments Using RNA Sequencing

In its natural habitat, the soil bacterium Bacillus subtilis often has to cope with fluctuating osmolality and nutrient availability. Upon nutrient depletion it can form dormant spores, which can revive to form vegetative cells when nutrients become available again. While the effects of salt stress on spore germination have been analyzed previously, detailed knowledge on the salt stress response during the subsequent outgrowth phase is lacking. In this study, we investigated the changes in gene expression during B. subtilis outgrowth in the presence of 1.2 M NaCl using RNA sequencing. In total, 402 different genes were upregulated and 632 genes were downregulated during 90 min of outgrowth in the presence of salt. The salt stress response of outgrowing spores largely resembled the osmospecific response of vegetative cells exposed to sustained high salinity and included strong upregulation of genes involved in osmoprotectant uptake and compatible solute synthesis. The σB-dependent general stress response typically triggered by salt shocks was not induced, whereas the σW regulon appears to play an important role for osmoadaptation of outgrowing spores. Furthermore, high salinity induced many changes in the membrane protein and transporter transcriptome. Overall, salt stress seemed to slow down the complex molecular reorganization processes (“ripening”) of outgrowing spores by exerting detrimental effects on vegetative functions such as amino acid metabolism

The Discovery of Gold Florins in Zalewo (formerly Saalfeld) in the Light of the Report from 1572

Artykuł dotyczy odkrycia skarbu złotych monet w Zalewie (dawniej Saalfeld). Książę Prus, Albrecht Fryderyk, został o nim poinformowany przez starostę przezmarskiego (Preussisch Mark), który wysłał mu 144 monety w 1572 r. Starosta opisał również okoliczności odkrycia. W kancelarii książęcej określono je jako florentinische Guldtgulden, tj. „florenckie złote guldeny”. Autorzy twierdzą, że jest to opis pierwszego odkrycia złotych florenów w Zalewie. Podobny skarb zarejestrowano tam w 1991 r. Monety wysłane w 1572 r. przypuszczalnie stały się częścią kolekcji księcia Prus. Niektóre z florenów odnalezionych w 1991 r. zostały przekazane Muzeum Warmii i Mazur.This article discusses the discovery of a hoard of gold coins in Zalewo (formerly Saalfeld). The duke of Prussia, Albrecht Frederick, was informed about the discovery by the Hauptmann of Przezmark (Preussisch Mark), who sent him 144 coins in 1572. The Hauptmann also described the process of how the coins were obtained. These coins were described by the archivist at the dukes‘s court as florentinische Guldtgulden – which means ”florentine gold florins“. The authors claim, that this is the first described discovery of gold florins from Zalewo. A similar discovery was recorded in Zalewo in 1991. The coins sent in 1572 possibly became a part of the collection of the Duke of Prussia. Some of the gold florins found in 1991 were donated to the Museum of Warmia and Mazury

New Source for the Location of the Manor of Teutonic Order Officials in Mortąg

Mortąg (dawne Mortung), to obecnie wieś położona w powiecie sztumskim (woj. pomorskie) wzmiankowana w źródłach po raz pierwszy w 1312 r. W XIV w. z Mortągiem związana jest siedziba prokuratora krzyżackiego (Pfleger / provisor) później mistrza rybickiego (Fischmeister). Urzędnicy krzyżaccy potwierdzeni są w Mortągu do 1455 r. Przy dworze urzędników w Mortągu funkcjonował także folwark, prokuratorom i mistrzom rybickim podlegały także wsie Stary Dzierzgoń (Alt Christburg), Monasterzysko (Munsterberg) oraz majątek i folwark Matule (Mothalen). W niniejszym artykule autorzy w oparciu o zachowane źródła wskazują potencjalne miejsce lokalizacji krzyżackich zabudowań związanych z funkcjonującym tu dworem oraz / lub folwarkiem.Mortąg (former Mortung), is presently a village situated in the county of Sztum (Pomeranian Voivodeship) mentioned in sources for the first time in 1312. In the 14th century the seat of the Teutonic Procurator (Pfleger / provisor) and later of the Master of Fishery (Fischmeister) is connected with Mortag. Teutonic officials are confirmed in Mortąg until 1455. There was also a manor farm (folwark) adjacent to the officials' manor in Mortag; the villages of Stary Dzierzgoń (Alt Christburg), Monasterzysko (Munsterberg) and the estate and manor of Matule (Mothalen) were also subject to the procurators and masters of fishery. In this article the authors, on the basis of the preserved sources, indicate the potential location of the Teutonic Knights' buildings connected with the manor and/or folwark functioning here

Propagation of scrapie in peripheral nerves after footpad infection in normal and neurotoxin exposed hamsters

As is known from various animal models, the spread of agents causing transmissible spongiform encephalopathies (TSE) after peripheral infection affects peripheral nerves before reaching the central nervous system (CNS) and leading to a fatal end of the disease. The lack of therapeutic approaches for TSE is partially due to the limited amount of information available on the involvement of host biological compartments and processes in the propagation of the infectious agent. The in vivo model presented here can provide information on the spread of the scrapie agent via the peripheral nerves of hamsters under normal and altered axonal conditions. Syrian hamsters were unilaterally footpad (f.p.) infected with scrapie. The results of the spatiotemporal ultrasensitive immunoblot-detection of scrapie-associated prion protein (PrP(Sc)) in serial nerve segments of both distal sciatic nerves could be interpreted as a centripetal and subsequent centrifugal neural spread of PrP(Sc) for this route of infection. In order to determine whether this propagation is dependent on main components in the axonal cytoskeleton (e.g. neurofilaments, also relevant for the component ;a' of slow axonal transport mechanisms), hamsters were treated -in an additional experiment- with the neurotoxin beta,beta-iminodiproprionitrile (IDPN) around the beginning of the scrapie infection. A comparison of the Western blot signals of PrP(Sc) in the ipsilateral and in the subsequently affected contralateral sciatic nerve segments with the results revealed from IDPN-untreated animals at preclinical and clinical stages of the TSE disease, indicated similar amounts of PrP(Sc). Furthermore, the mean survival time was unchanged in both groups. This in vivo model, therefore, suggests that the propagation of PrP(Sc) along peripheral nerves is not dependent on an intact neurofilament component of the axonal cytoskeleton. Additionally, the model indicates that the spread of PrP(Sc) is not mediated by the slow component ;a' of the axonal transport mechanism

Phytochrome and endogenous gibberellin-like substances in etiolated and irradiated oat seedlings

The level of gibberellin-like substances was investigated in oat coleoptiles with different stationary states of phytochrome and in leaf segments which had been etiolated and irradiated with red light. Etiolated coleoptiles and leaves containing maximum amounts of the PR form of phytochrome were characterized by an increased level of bound gibberellins. Irradiation with red light resulting in the photoconversion of phytochrome into the Pt R form caused the appearance of a high content of free gibberellins. It seems that the releasing the hormones from bound forms correlated with the formation of phytochrome PFR may be an important aspect of the mechanism of phytochrome action in the processes of seedling deetiolation. The interrelation between phytochrome and plant hormones in the control of photomorphogenesis of young monocotyledonous seedlings is also discussed

Easy and cost-effective stable positioning of suspension cells during live-cell imaging

Dynamic processes of cells can be best monitored when living cells are analyzed by imaging. While it is easy to observe adherent living cells it has been extremely challenging to analyze suspension cells. This cell type floats freely in the culture dish, and it is only a question of time when the focus or the observation field is lost. In order to keep the cells in focus, an easy and inexpensive method allowing the observation of living suspension cells during confocal laser scanning microscope imaging was developed

tANCHOR: A Novel Mammalian Cell Surface Peptide Display System

A novel tool for the presentation of peptides and small proteins on the surface of human cells has been developed. Our tANCHOR system utilizes tetraspanin anchors containing heterologous amino acid sequences inserted instead of the large extracellular loop. This technology allows a highly effective extracellular display of epitopes for antibody binding studies and many other potential applications.Peer Reviewe