Francisella tularensis elicits IL-10 via a PGE₂-inducible factor, to drive macrophage MARCH1 expression and class II down-regulation

Francisella tularensis is a bacterial pathogen that uses host-derived PGE? to subvert the host's adaptive immune responses in multiple ways. Francisella-induced PGE? acts directly on CD4 T cells to blunt production of IFN-γ. Francisella-induced PGE? can also elicit production of a >10 kDa soluble host factor termed FTM?SN (F. tularensismacrophage supernatant), which acts on IFN-γ pre-activated M? to down-regulate MHC class II expression via a ubiquitin-dependent mechanism, blocking antigen presentation to CD4 T cells. Here, we report that FTM?SN-induced down-regulation of M? class II is the result of the induction of MARCH1, and that M? expressing MARCH1 "resistant" class II molecules are resistant to FTM?SN-induced class II down-regulation. Since PGE? can induce IL-10 production and IL-10 is the only reported cytokine able to induce MARCH1 expression in monocytes and dendritic cells, these findings suggested that IL-10 is the active factor in FTM?SN. However, use of IL-10 knockout M? established that IL-10 is not the active factor in FTM?SN, but rather that Francisella-elicited PGE? drives production of a >10 kDa host factor distinct from IL-10. This factor then drives M? IL-10 production to induce MARCH1 expression and the resultant class II down-regulation. Since many human pathogens such as Salmonella typhi, Mycobacterium tuberculosis and Legionella pneumophila also induce production of host PGE?, these results suggest that a yet-to-be-identified PGE?-inducible host factor capable of inducing IL-10 is central to the immune evasion mechanisms of multiple important human pathogens.     

A new series of pyridinyl-alkynes as antagonists of the metabotropic glutamate receptor 5 (mGluR5)

Synthesis and some structure-activity relationships for a new series of propargyl ethers as mGluR5 antagonists are reported.     

Sapwood estimates of pedunculate oak (Quercus robur L.) in eastern Baltic

Pedunculate oak (Quercus robur L.) is one of the widely used and dendrochronologically investigated species in Europe. Still, it is a problematical dating object if its outermost section is missing partly or totally. Thus, we need sapwood estimation of living trees. As sapwood amount varies geographically, numbers of sapwood rings have been published for different regions in Europe but no such estimation has been done for the Baltic States yet. Therefore, this paper deals with the estimation of pedunculate oak sapwood growing in the eastern Baltic region, i.e. in Finland, Estonia, Latvia, and Lithuania.In total, 668 oak core samples of living trees from 43 stands were investigated. Ring widths were measured and the number of sapwood rings was determined according to two criteria: difference of colour and absence of tyloses in earlywood vessels. The samples were divided into two sets, according to the t_H-values between site chronologies and the major geobotanical sub-provinces. Thus, the nine Finnish and western Estonian sites were attributed to the western region and the 34 eastern Estonian, Latvian and Lithuanian sites to the eastern region.As the result of a statistical analysis, we explain that the number of oak sapwood rings ranges from 4.09 to 20.85 and 6.45 to 18.02 within 95% confidence limits in the western and eastern regions, respectively. For the three Baltic countries and southern Finland in general, we recommend to consider a sapwood estimate of 6.18–18.71 rings. Regarding earlier studies, the general European trend of decreasing sapwood ring number towards the east was confirmed. A geographical pattern of eastward decrease of the median sapwood ring number was noticed in the Baltics as well. The chronology based upon 668 samples of living oak trees from all sites covered the period of 1631–2008.     

Ablation of RIC8A Function in Mouse Neurons Leads to a Severe Neuromuscular Phenotype and Postnatal Death

Resistance to inhibitors of cholinesterase 8 (RIC8) is a guanine nucleotide exchange factor required for the intracellular regulation of G protein signalling. RIC8 activates different Gα subunits via non-canonical pathway, thereby amplifying and prolonging the G protein mediated signal. In order to circumvent the embryonic lethality associated with the absence of RIC8A and to study its role in the nervous system, we constructed Ric8a conditional knockout mice using Cre/loxP technology. Introduction of a synapsin I promoter driven Cre transgenic mouse strain (SynCre) into the floxed Ric8a (Ric8a^F/F) background ablated RIC8A function in most differentiated neuron populations. Mutant SynCre^+/-Ric8^lacZ/F mice were born at expected Mendelian ratio, but they died in early postnatal age (P4-P6). The mutants exhibited major developmental defects, like growth retardation and muscular weakness, impaired coordination and balance, muscular spasms and abnormal heart beat. Histological analysis revealed that the deficiency of RIC8A in neurons caused skeletal muscle atrophy and heart muscle hypoplasia, in addition, the sinoatrial node was misplaced and its size reduced. However, we did not observe gross morphological changes in brains of SynCre^+/-Ric8a^lacZ/F mutants. Our results demonstrate that in mice the activity of RIC8A in neurons is essential for survival and its deficiency causes a severe neuromuscular phenotype.     

Expression Pattern and Localization Dynamics of Guanine Nucleotide Exchange Factor RIC8 during Mouse Oogenesis

Targeting of G proteins to the cell cortex and their activation is one of the triggers of both asymmetric and symmetric cell division. Resistance to inhibitors of cholinesterase 8 (RIC8), a guanine nucleotide exchange factor, activates a certain subgroup of G protein α-subunits in a receptor independent manner. RIC8 controls the asymmetric cell division in Caenorhabditis elegans and Drosophila melanogaster, and symmetric cell division in cultured mammalian cells, where it regulates the mitotic spindle orientation. Although intensely studied in mitosis, the function of RIC8 in mammalian meiosis has remained unknown. Here we demonstrate that the expression and subcellular localization of RIC8 changes profoundly during mouse oogenesis. Immunofluorescence studies revealed that RIC8 expression is dependent on oocyte growth and cell cycle phase. During oocyte growth, RIC8 is abundantly present in cytoplasm of oocytes at primordial, primary and secondary preantral follicle stages. Later, upon oocyte maturation RIC8 also populates the germinal vesicle, its localization becomes cell cycle dependent, and it associates with chromatin and the meiotic spindle. After fertilization, RIC8 protein converges to the pronuclei and is also detectable at high levels in the nucleolus precursor bodies of both maternal and paternal pronucleus. During first cleavage of zygote RIC8 localizes in the mitotic spindle and cell cortex of forming blastomeres. In addition, we demonstrate that RIC8 co-localizes with its interaction partners Gα_i1/2:GDP and LGN in meiotic/mitotic spindle, cell cortex and polar bodies of maturing oocytes and zygotes. Downregulation of Ric8 by siRNA leads to interferred translocation of Gα_i1/2 to cortical region of maturing oocytes and reduction of its levels. RIC8 is also expressed at high level in female reproductive organs e.g. oviduct. Therefore we suggest a regulatory function for RIC8 in mammalian gametogenesis and fertility.     

Multi-Parametric MRI and Texture Analysis to Visualize Spatial Histologic Heterogeneity and Tumor Extent in Glioblastoma

Background

Genetic profiling represents the future of neuro-oncology but suffers from inadequate biopsies in heterogeneous tumors like Glioblastoma (GBM). Contrast-enhanced MRI (CE-MRI) targets enhancing core (ENH) but yields adequate tumor in only ~60% of cases. Further, CE-MRI poorly localizes infiltrative tumor within surrounding non-enhancing parenchyma, or brain-around-tumor (BAT), despite the importance of characterizing this tumor segment, which universally recurs. In this study, we use multiple texture analysis and machine learning (ML) algorithms to analyze multi-parametric MRI, and produce new images indicating tumor-rich targets in GBM.

Methods

We recruited primary GBM patients undergoing image-guided biopsies and acquired pre-operative MRI: CE-MRI, Dynamic-Susceptibility-weighted-Contrast-enhanced-MRI, and Diffusion Tensor Imaging. Following image coregistration and region of interest placement at biopsy locations, we compared MRI metrics and regional texture with histologic diagnoses of high- vs low-tumor content (≥80% vs <80% tumor nuclei) for corresponding samples. In a training set, we used three texture analysis algorithms and three ML methods to identify MRI-texture features that optimized model accuracy to distinguish tumor content. We confirmed model accuracy in a separate validation set.

Results

We collected 82 biopsies from 18 GBMs throughout ENH and BAT. The MRI-based model achieved 85% cross-validated accuracy to diagnose high- vs low-tumor in the training set (60 biopsies, 11 patients). The model achieved 81.8% accuracy in the validation set (22 biopsies, 7 patients).

Conclusion

Multi-parametric MRI and texture analysis can help characterize and visualize GBM’s spatial histologic heterogeneity to identify regional tumor-rich biopsy targets.     

Common growth signal and spatial synchrony of the chronologies of tree-rings from pines in the Baltic Sea region over the last nine centuries

The common growth signal and spatial synchrony of nine chronologies of tree rings (Pinus sylvestris L.) from the Baltic Sea region were studied using moving correlations with time windows of 50 and 100 years. Drifts from synchrony to asynchrony and again back to synchrony across several centuries were observed. The chronologies showed higher (or lower) coefficients of correlations and correspondingly gentler (or steeper) declines in spatial synchrony in certain periods. In accordance with research into other ecological phenomena, the origin of the spatial synchrony was found to relate to the intensity of the westerly circulation over the Atlantic and Europe as described by the North Atlantic Oscillation (NAO) index. The tree rings from all of the analysed subregions were found to be wider, identifying faster periods of growth, when there was a positive NAO during the extended winter season from December to March that preceded the period of growth. Previous studies showed that the NAO index can be linked with the growth of tree rings because of factors in the local climate that affect growth, such as temperature and precipitation. Moreover, the spatial synchrony showed multidecadal fluctuations that were correlated with variations of a similar scale in the reconstructed NAO index. However, a high degree of spatial synchrony was observed during the 20^th century in the presence of values of NAO index that were strongly negative. A more detailed view was provided by maps of the correlations in which spatial changes were demonstrated in the growth of pines between the 12^th and the 20^th century. These results could be used as background information in the development of improved proxy-based reconstructions of the NAO index (or other similar meteorological variables), which could also be inferred from tree ring data relating to earlier centuries.     

Temperature regime of planted roofs compared with conventional roofing systems

This paper analyses the temperature regime of an existing green roof and a sod roof, compared with a modified bituminous membrane roof and a steel sheet roof. The measurement period was from June 2004 to December 2007 at three different measurement locations. Results are given both seasonally and daily; indexes to characterize the effects of the temperature of planted roofs are also proposed. In summer, temperatures under both the green roof (100 mm) and the sod roof (150 mm) showed a similar temperature run; undesirable higher temperatures on the surfaces did not cause a notable increase in temperature under the substrate layers. The difference between the temperature amplitude under the substrate layers of the planted roofs and the surfaces of the conventional roofs was on average 20 °C. In autumn and spring, the sod roof's soil layer showed higher temperatures and lower amplitude than the green roof's substrate layer, which cooled more. In winter, temperatures under the substrate layers of the planted roofs were higher than the surfaces of the conventional roofs; average amplitude was 1 °C and 7–8 °C, respectively.