EphB receptors coordinate migration and proliferation in the intestinal stem cell niche

More than 10(10) cells are generated every day in the human intestine. Wnt proteins are key regulators of proliferation and are known endogenous mitogens for intestinal progenitor cells. The positioning of cells within the stem cell niche in the intestinal epithelium is controlled by B subclass ephrins through their interaction with EphB receptors. We report that EphB receptors, in addition to directing cell migration, regulate proliferation in the intestine. EphB signaling promotes cell-cycle reentry of progenitor cells and accounts for approximately 50% of the mitogenic activity in the adult mouse small intestine and colon. These data establish EphB receptors as key coordinators of migration and proliferation in the intestinal stem cell niche.     

Towards hemerythrin-based blood substitutes: Comparative performance to hemoglobin on human leukocytes and umbilical vein endothelial cells

Hemerythrin is a dioxygen-carrying protein whose oxidative/nitrosative stress-related reactivity is lower than that of hemoglobin, which may warrant investigation of hemerythrin as raw material for artificial oxygen carriers (‘blood substitutes’). We report here the first biological tests for hemerythrin and its chemical derivatives, comparing their performance with that of a representative competitor, glutaraldehyde-polymerized bovine hemoglobin. Hemerythrin (native or derivatized) exhibits a proliferative effect on human umbilical vein endothelial cell (HUVEC) cultures, as opposed to a slight inhibitory effect of hemoglobin. A similar positive effect is displayed on human lymphocytes by glutaraldehyde-polymerized hemerythrin, but not by native or polyethylene glycol-derivatized hemerythrin.     

Sema4C-Plexin B2 signalling modulates ureteric branching in developing kidney

Semaphorins, originally identified as axon guidance molecules, have also been implicated in angiogenesis, function of the immune system and cancerous growth. Here we show that deletion of Plexin B2 (Plxnb2), a semaphorin receptor that is expressed both in the pretubular aggregates and the ureteric epithelium in the developing kidney, results in renal hypoplasia and occasional double ureters. The rate of cell proliferation in the ureteric epithelium and consequently the number of ureteric tips are reduced in the kidneys lacking Plexin B2 (Plxnb2-/-). Semaphorin 4C, a ligand for Plexin B2, stimulates branching of the ureteric epithelium in wild type and Plxnb2+/- kidney explants, but not in Plxnb2-/- explants. As shown by co-immunoprecipitation Plexin B2 interacts with the Ret receptor tyrosine kinase, the receptor of Glial-cell-line-derived neurotrophic factor (Gdnf), in embryonic kidneys. Isolated Plxnb2-/- ureteric buds fail to respond to Gdnf by branching, but this response is rescued by Fibroblast growth factor 7 and Follistatin as well as by the metanephric mesenchyme. The differentiation of the nephrogenic mesenchyme, its morphology and the rate of apoptosis in the Plxnb2-/- kidneys are normal. Plexin B2 is co-expressed with Plexin B1 (Plxnb1) in the kidney. The double homozygous Plxnb1-Plxnb2-deficient mice show high embryonic lethality prior to onset of nephrogenesis. The only double homozygous embryo surviving to E12 showed hypoplastic kidneys with ureteric branches and differentiating mesenchyme. Taken together, our results show that Sema4C-Plexin B2 signalling regulates ureteric branching, possibly through modulation of Gdnf signalling by interaction with Ret, and suggest non-redundant roles for Plexin B1 and Plexin B2 in kidney development.     

Ungulate browsing shapes climate change impacts on forest biodiversity in Hungary

Climate change can result in a slow disappearance of forests dominated by less drought-tolerant native European beech (Fagus sylvatica) and oak species (Quercus spp.) and further area expansion of more drought-tolerant non-native black locust (Robinia pseudoacacia) against those species in Hungary. We assumed that the shift in plant species composition was modified by selective ungulate browsing. Thus, we investigated which woody species are selected by browsing game. We have collected data on the species composition of the understory and the browsing impact on it in five different Hungarian even-aged forests between 2003 and 2005. Based on these investigations the non-native Robinia pseudoacacia living under more favourable climatic conditions was generally preferred (Jacobs’ selectivity index: D = 0.04 ± 0.77), while the native Fagus sylvatica and Quercus spp. (Q. petraea, Q. robur), both more vulnerable to increasing aridity, were avoided (D = ?0.37 ± 0.11; ?0.79 ± 0.56; ?0.9 ± 0.16; respectively) among target tree species. However, economically less or not relevant species, e.g. elderberry (Sambucus spp.), blackberry (Rubus spp.) or common dogwood (Cornus sanguinea) were the most preferred species (D = 0.01 ± 0.71; ?0.12 ± 0.58; ?0.2 ± 0.78, respectively). Our results imply that biodiversity conservation, i.e. maintaining or establishing a multi-species understory layer, can be a good solution to reduce the additional negative game impact on native target tree species suffering from drought. Due to preference for Robinia pseudoacacia selective browsing can decelerate the penetration of this species into native forest habitats. We have to consider the herbivorous pressure of ungulates and their feeding preferences in planning our future multifunctional forests in the light of climate change impacts.     

TrAp: a tree approach for fingerprinting subclonal tumor composition

Revealing the clonal composition of a single tumor is essential for identifying cell subpopulations with metastatic potential in primary tumors or with resistance to therapies in metastatic tumors. Sequencing technologies provide only an overview of the aggregate of numerous cells. Computational approaches to de-mix a collective signal composed of the aberrations of a mixed cell population of a tumor sample into its individual components are not available. We propose an evolutionary framework for deconvolving data from a single genome-wide experiment to infer the composition, abundance and evolutionary paths of the underlying cell subpopulations of a tumor. We have developed an algorithm (TrAp) for solving this mixture problem. In silico analyses show that TrAp correctly deconvolves mixed subpopulations when the number of subpopulations and the measurement errors are moderate. We demonstrate the applicability of the method using tumor karyotypes and somatic hypermutation data sets. We applied TrAp to Exome-Seq experiment of a renal cell carcinoma tumor sample and compared the mutational profile of the inferred subpopulations to the mutational profiles of single cells of the same tumor. Finally, we deconvolve sequencing data from eight acute myeloid leukemia patients and three distinct metastases of one melanoma patient to exhibit the evolutionary relationships of their subpopulations.     

Epidermal Growth Factor and Basic Fibroblast Growth Factor Have Independent Actions on Mesencephalic Dopamine Neurons in Culture

Abstract: Epidermal growth factor (EGF) and basic fibro-blast growth factor (bFGF) are both trophic for dopamine neurons s in cultures of dissociated embryonic rat mesen-cephaion, but the significance of this apparent overlap in neurotrophic activity is not yet known. In this study, we investigated the mechanisms of action of these two growth factors and the potential relationship between them, Using a nuclease protection assay, we determined that bFGF mRNA was expressed in the cultures. Double-label immunocytochemistry revealed that bFGF immunore-active material could be detected in tyrosine hydroxylase immunoreactive neurons and glial fibrillary acidic protein immunoreactive astrocytes. EGF treatment increased bFGF mRNA content per culture dish. As we have previously demonstrated that EGF exerts its dopaminergic neurotrophic activity via an intermediate cell type, studies were designed to address whether the pathway by which EGF acts on dopaminergic neurons is mediated by the release of bFGF. However, the trophic action of EGF on dopamine neurons, represented by high-affinity neuronal dopamine uptake, could not be blocked by immunoneutralization of bFGF, suggesting that the actions of EGF were not mediated by bFGF release. The time course of the effects of EGF and bFGF on dopamine uptake were similar, with significant increases detectable only after 5 days in culture. Both growth factors were active in the picomolar-to-nannomolar range with maximal trophic activity between 0.4 and 2.5 n M. EGF, however, was the more potent mitogen under these conditions. When cultures were simultaneously incubated with maximal concentrations of EGF and bFGF, the effect on dopamine uptake was significantly greater than with either growth factor alone and, in fact, approximated the sum of the individual effects. On the basis of these results we conclude that these growth factors have independent effects on dopamine neurons of the mesencephalon.     

Synthesis of Polyfused Heteroaromatics by Palladium-Catalyzed Cross-Coupling Reaction

The palladium-catalyzed coupling of unsaturated halides or triflates with organostannanes (the Stille reaction) or with organo-boronic acids (the Suzuki reaction) has envolved as a powerful means of carbon-carbon bond formation. As an extention of our work on the synthesis and reactivity of positively charged bridge-head nitrogen containing fused azinium salts, we have been studying the formation of polyfused heteroaromatics with angular anellation pattern. The reaction of such positively charged salts with nucleophiles has proven to be a suitable tool for the preparation of heteroaryl dienes, which have been shown to be excellent intermediates for several ring closure reaction. A selection of results will be presented, including applications to natural product synthesis.     

Release of adenosine from human neutrophils stimulated by platelet activating factor, leukotriene B(4) and opsonized zymosan

Isolated human polymorphonuclear leukocytes (PMNL) stimulated by platelet activating factor (PAF), leukotriene B(4) (LTB(4)) or opsonized zymosan (OZ) released adenosine measured by thermospray high performance liquid chromatography mass spectrometry in the cell-free supernatants. Stimulation by PAF or LTB(4) resulted in a bellshaped concentration-effect curve; 5 x 10(-7) M PAF, 10(-8) M LTB(4) and 500 mug ml(-1) OZ induced peak adenosine release, thus cytotoxic concentrations did not elevate adenosine level in the supernatants. Therefore adenosine release was characteristic of viable cells. As calculated from concentration-effect curves, the rank order of potency for adenosine release was PAF > LTB > OZ. These resuits suggest that adenosine, when bound specifically to membrane receptor sites, may initiate signal transduction, and, in co-operation with other inflammatory mediators, may modulate phagocyte function, e.g. production of chemoluminescence (CL).