In bacteria, the so-called stringent response is responsible for adaptation to changing environmental conditions. This response is mediated by guanosine derivatives [(p)ppGpp], synthesized by either large mono-functional RelA or bi-functional SpoT (synthesis and hydrolysis) enzymes in β- and γ-proteobacteria, such as Escherichia coli. In Firmicutes and α-, δ- and ε-proteobacteria, large bifunctional Rel-SpoT-Homologs (RSH), often accompanied by small (p)ppGpp synthetases and/or hydrolases devoid of regulatory domains, are responsible for (p)ppGpp turnover. Here, we report on surprising in vitro and in vivo properties of an RSH enzyme from Methylobacterium extorquens (RSHMex). We find that this enzyme possesses some unique features, e.g. it requires cobalt cations for the most efficient (p)ppGpp synthesis, in contrast to all other known specific (p)ppGpp synthetases that require Mg2+. In addition, it can synthesize pppApp, which has not been demonstrated in vitro for any Rel/SpoT/RSH enzyme so far. In vivo, our studies also show that RSHMex is active in E. coli cells, as it can complement E.coli ppGpp0 growth defects and affects rrnB P1-lacZ fusion activity in a way expected for an RSH enzyme. These studies also led us to discover pppApp synthesis in wild type E.coli cells (not carrying the RSHMex enzyme), which to our knowledge has not been demonstrated ever before. In the light of our recent discovery that pppApp directly regulates E. coli RNAP transcription in vitro in a manner opposite to (p)ppGpp, this leads to a possibility that pppApp is a new member of the nucleotide second-messenger family that is widely present in bacterial species.

Heat shock proteins (Hsp) are constitutive and stress-induced molecules which have been reported to impact innate and adaptive immune responses. Here, we evaluated the role of Hsp70 as a treatment target in the imiquimod-induced, psoriasis-like skin inflammation mouse model and related in vitro assays. We found that immunization of mice with Hsp70 resulted in decreased clinical and histological disease severity associated with expansion of T cells in favor of regulatory subtypes (CD4⁺FoxP3⁺/CD4⁺CD25⁺ cells). Similarly, anti-Hsp70 antibody treatment led to lowered disease activity associated with down-regulation of pro-inflammatory Th17 cells. A direct stimulating action of Hsp70 on regulatory T cells and its anti-proliferative effects on keratinocytes were confirmed in cell culture experiments. Our observations suggest that Hsp70 may be a promising therapeutic target in psoriasis and potentially other autoimmune dermatoses.

Extracellular heat shock proteins (Hsp) influence the adaptive immune response and may ameliorate pathogenesis of autoimmune diseases. While some preclinical observations suggest that highly conserved bacterial and/or murine Hsp70 peptides have potential utility in treatment of rheumatoid arthritis (RA) via induction of T regulatory cells (Treg), the role of extracellular inducible human Hsp70 in adaptive immune processes requires further investigation. The present study evaluated Hsp70 influence on inflammatory cytokine-mediated modulation of T cell immunophenotype in ways that influence RA onset and severity. Initial experiments in the present investigation revealed that serum levels of Hsp70 are approximately 2-fold higher in RA patients versus healthy control subjects. To explore the effect of extracellular Hsp70 on key processes underlying the adaptive immune system, the effects of a highly pure, substrate-, and endotoxin-free human Hsp70 on polarization of the T helper cell subpopulations, including CD4⁺IL-17⁺ (Th17), CD4⁺FoxP3⁺ (Treg), CD4⁺IFN-γ⁺ (Th1), and CD4⁺IL-4⁺ (Th2), were studied in naïve human peripheral blood mononuclear cell (PBMC) cultures stimulated with anti-CD3/28 mAb. Major findings included an observation that while Hsp70 treatment increased Th17 frequencies and Th17/Treg ratio, the frequency of Th1 cells and the Th1/Th2 ratio were significantly decreased in the Hsp70-treated PBMC cultures. Moreover, data shown here provides preliminary suggestion that major contributing Hsp70-mediated immunomodulation includes interleukin 6 (IL-6) influence on Th17/Treg and Th1/Th2, since expression of this inflammatory cytokine is enhanced by in vitro Hsp70 treatment. These results are nevertheless preliminary and require further investigation to validate the above model.

Many landscapes bear the marks of historical land use. These marks can be the basis for a reconstruction of a historical land use structure as some of them are typical of different types of human activity. The aim of this paper is to determine whether Austrian cadastral maps from the 19th century present the image of the most transformed environment in the Western Carpathians as a result of agricultural activity. Land use structure and terrain forms were detected based on Austrian cadastral maps from 1848, airborne laser scanning and field studies. In two of the test areas, the percentage of arable fields was higher among the plots with stone mounds than the percentage among the plots without them. In the third test area, the relationship was reversed. Also, lynchets, terraces and stone walls sometimes occur in plots that were not arable fields in 1848. Thus, the Austrian cadastral maps from 1848 could not reflect the maximal range of arable fields in the Carpathians in the 19th century. However, it is impossible to determine the historical structure of land use precisely. Nevertheless, an inventory of terrain forms can be used to assess land use when historical maps have not preserved or when available maps do not present land use in detail.

The recent increase in urban areas has stimulated landscape urbanization. One of the ways to study this process is an analysis based on the structure of land cover. The aim of this paper is to assess the intensity of the urban landscape on the basis of the CORINE in the seven largest metropolitan areas in Poland and in the Ruhr Metropolis in Germany. To this end, an urban landscape intensity indicator (ULII) was used based on Corine Land Cover at three levels of detail: the metropolitan area, municipalities and hexagons. There are similarities in landscape structure in areas with similar origin (industrial function) and spatial organization (mono- and polycentric agglomerations). The landscape of the Upper Silesia-Zagłębie Metropolis differs from the landscape of other metropolitan areas in Poland and simultaneously shows similarities to the landscape of the Ruhr Metropolis. The results of the ULII also revealed a dependency: the dominance of rural and transitional landscapes in a majority of the study areas. Urban landscapes occur only in the central zones of the metropolitan areas. This proves that determining the range of a metropolitan area in terms of landscape factors is different from doing it with formal or legal ones.