5-Keto-3-cyano-2,4-diaminothiophenes as selective maternal embryonic leucine zipper kinase inhibitors

Maternal embryonic leucine zipper kinase (MELK) is involved in several key cellular processes and displays increased levels of expression in numerous cancer classes (colon, breast, brain, ovary, prostate and lung). Although no selective MELK inhibitors have yet been approved, increasing evidence suggest that inhibition of MELK would constitute a promising approach for cancer therapy. A weak high-throughput screening hit (17, IC₅₀?≈?5?μM) with lead-like properties was optimized for MELK inhibition. The early identification of a plausible binding mode by molecular modeling offered guidance in the choice of modifications towards compound 52 which displayed a 98?nM IC₅₀. A good selectivity profile was achieved for a representative member of the series (29) in a 486 protein kinase panel. Future elaboration of 52 has the potential to deliver compounds for further development with chemotherapeutic aims.     

Pescadillo homologue 1 and Peter Pan function during Xenopus laevis pronephros development

Background information. pes1 (pescadillo homologue 1) and ppan (Peter Pan) are multifunctional proteins involved in ribosome biogenesis, cell proliferation, apoptosis, cell migration and regulation of gene expression. Both proteins are required for early neural development in Xenopus laevis, as previously demonstrated. Results. We show that the expression of both genes in the developing pronephros depends on wnt4 and fzd3 (frizzled homologue 3) function. Loss of pes1 or ppan by MO (morpholino oligonucleotide)‐based knockdown approaches resulted in strong malformations during pronephric tubule formation. Defects were already notable during specification of pronephric progenitor cells, as shown by lhx1 expression. Moreover, we demonstrated that Xenopus pes1 and ppan interact physically and functionally and that pes1 and ppan can cross‐rescue the loss of function phenotype of one another. Interference with rRNA synthesis, however, did not result in a similar early pronephros phenotype. Conclusion. These results demonstrate that pes1 and ppan are required for Xenopus pronephros development and indicate that their function in the pronephros is independent of their role in ribosome biosynthesis.     

JAK2 mutation status, hemostatic risk factors and thrombophilic factors in essential thrombocythemia (ET) patients

The recently discovered JAK2 V617F point mutation, found in 50-60% of ET patients, has been reported to be associated with a higher risk of thrombotic events. In this study, we explored if JAK2 V617F mutation, or coexisting thrombophilic and hemostatic risk factors, contributed to these complications. We examined 32 patients with ET, and looked for pathogenetic JAK2 V617F mutation and prothrombotic genes mutations: factor V Leiden, prothrombin and MTHFR. We also evaluated plasma levels of fibrinogen, factors VIII and XII, AT, protein C, protein S and serum level of homocysteine. Urokinase concentration was assessed in patients' plasma as well as platelet lysates. There was no difference in the number of thrombotic complications between ET patients with and without JAK2 mutation. However, we found a number of thrombophilic and hemostatic risk factors that could contribute to thrombotic complications in ET patients.     

Coveting thy neighbors fitness as a means to resolve social dilemmas

In spatial evolutionary games the fitness of each individual is traditionally determined by the payoffs it obtains upon playing the game with its neighbors. Since defection yields the highest individual benefits, the outlook for cooperators is gloomy. While network reciprocity promotes collaborative efforts, chances of averting the impending social decline are slim if the temptation to defect is strong. It is, therefore, of interest to identify viable mechanisms that provide additional support for the evolution of cooperation. Inspired by the fact that the environment may be just as important as inheritance for individual development, we introduce a simple switch that allows a player to either keep its original payoff or use the average payoff of all its neighbors. Depending on which payoff is higher, the influence of either option can be tuned by means of a single parameter. We show that, in general, taking into account the environment promotes cooperation. Yet coveting the fitness of one's neighbors too strongly is not optimal. In fact, cooperation thrives best only if the influence of payoffs obtained in the traditional way is equal to that of the average payoff of the neighborhood. We present results for the prisoner's dilemma and the snowdrift game, for different levels of uncertainty governing the strategy adoption process, and for different neighborhood sizes. Our approach outlines a viable route to increased levels of cooperative behavior in structured populations, but one that requires a thoughtful implementation.     

Molecular investigation of enteroaggregative, shiga toxin-producing E. coli O104:H4 isolated in Poland during the recent international outbreak--characteristic of epidemic clone

Since early May 2011 a large food-borne outbreak caused by E. coli O104:H4 affected Germany then spread over 13 European countries, U.S.A. and Canada. The outbreak strain was found to possess an unusual combination of enteroaggregative E. coli pathotype with StxII. In this report we described the molecular investigation of epidemic clone in Poland during the international outbreak. We confirmed three cases of E. coli O104:H4 infections. The molecular characteristics of the Polish E. coli O104:H4 isolates including virulence profile, antimicrobial resistance, PFGE and plasmids profiles were corresponded with Germany outbreak strains.     

Assessment of essential oils activity used in liquid or volatile phase against biofilm cultures of Staphylococcus aureus and Candida albicans. The methodological study

Biofilm formation is a significant factor in chronic infections with fungal and bacterial pathogens. Due to the high drug resistance of biofilm populations and the frequent failures of chemotherapy i such infections, it seems necessary to take recourse to unconventional treatment methods involving e.g. the use of some phytocompounds such as essential oils or their components. In order to evaluate the effect of their action on the microbial biomass a variety of techniques are used. However, there is still a need to develop new tests or modifications of these known, for the biofilm viability assessment. They should be adapted to the physico-chemical nature of the tested compounds and should decrease the risk ofbiofilm damage during staining procedure. We described a test assessing the effect of essential oils on bacterial and fungal biofilm formed on the membrane of cell culture inserts. The proposed model provides a minimal violation of the biofilm integrity during the test. It allows easily explore the activity of essential oil volatile fraction and is useful in determination of the kinetics of their action. Using this test it is also easy to examine the relationship between antimicrobial activity and the cytotoxic effect, known as the biocompatibility index (BI, biocompatibility index). Moreover, it allows qualitative and quantitative analysis of metabolic products, released into the growth medium from biofilm's cells. In successively repeated experiments high reproducibility of results has been obtained, thus the developed methodology seems to be useful in our future studies in this field.     

Reliability of the cut-off value in the routine serodiagnosis of pertussis performed by the commercial ELISA assays

In clinical practise, serodiagnosis of pertussis is mostly based on single-sample serology using a single cut-off. The reliability of the cut-off value has the crucial influence for the sensitivity and specificity of the used tests. In this context we compared the value of cut-off used in two commercial ELISA kits (NovaLisa Bordetella pertussis-NovaTec and ELISA Bordetella pertussis ELISA-Virotech) with cut-off settled by calculation the IgA and IgG results from the 60 healthy Polish children and 100 blood donors (arithmetic mean plus 2 or 3 standard deviations). Our study indicates that IgA cutoff used in NovaTec ELISA, in contrary to Virotech ELISA, correspond better to the level x+3SD calculated in children sera and x+2SD calculated in adult blood donors sera. The value of IgG cut-off used in Virotech ELISA was lower about 20% and 30% from the cut-off settled by us respectively on the level ofx+2SD and x+3SD in all tested sera. The most inadequate value had the IgG cut-offused in NovaTec ELISA, which was over three times lower than mean cut-off value settled by calculation results from the sera obtained from healthy children and blood donors. This low cut-off value established by the NovaTec was the reason that 23.3% of healthy children and 55.0% of blood donors have the IgG antibodies on the diagnostic significant level. Our data suggest that commercial ELISAs need further improvement and standardization.     

Proteomic characterization of archaeal ribosomes reveals the presence of novel archaeal-specific ribosomal proteins

Protein synthesis occurs in macromolecular particles called ribosomes. All ribosomes are composed of RNA and proteins. While the protein composition of bacterial and eukaryotic ribosomes has been well-characterized, a systematic analysis of archaeal ribosomes has been lacking. Here we report the first comprehensive two-dimensional PAGE and mass spectrometry analysis of archaeal ribosomes isolated from the thermophilic Pyrobaculum aerophilum and the thermoacidophilic Sulfolobus acidocaldarius Crenarchaeota. Our analysis identified all 66 ribosomal proteins (r-proteins) of the P. aerophilum small and large subunits, as well as all but two (62 of 64; 97%) r-proteins of the S. acidocaldarius small and large subunits that are predicted genomically. Some r-proteins were identified with one or two lysine methylations and N-terminal acetylations. In addition, we identify three hypothetical proteins that appear to be bona fide r-proteins of the S. acidocaldarius large subunit. Dissociation of r-proteins from the S. acidocaldarius large subunit indicates that the novel r-proteins establish tighter interactions with the large subunit than some integral r-proteins. Furthermore, cryo electron microscopy reconstructions of the S. acidocaldarius and P. aerophilum 50S subunits allow for a tentative localization of the binding site of the novel r-proteins. This study illustrates not only the potential diversity of the archaeal ribosomes but also the necessity to experimentally analyze the archaeal ribosomes to ascertain their protein composition. The discovery of novel archaeal r-proteins and factors may be the first step to understanding how archaeal ribosomes cope with extreme environmental conditions.     

In vitro cultures of Drosera aliciae as a source of a cytotoxic naphthoquinone: ramentaceone

A protocol for the in vitro propagation of Drosera aliciae to increase the yield of the naphthoquinone, ramentaceone, was developed. The highest micropropagation coefficient was obtained using half-strength Murashige–Skoog medium supplemented with 0.4 μM 6-benzyladenine (BA). The genetic fidelity and stability of the regenerated plants was confirmed with RAPD markers. The activity of the isolated ramentaceone was determined against four human tumor cell lines: U937, HeLa, MCF-7, HCT-116 with the highest cytotoxic activity towards the leukemic U937 cell line with an IC₅₀ value of 3.2 μM.     

Design and synthesis of CK2 inhibitors

A series of new polybrominated benzimidazoles and benzotriazoles has been synthesized and their influence on the activity of protein kinase CK2 was evaluated. It was revealed that the most active inhibitors are those with methyl or ethyl substituent at benzene ring, namely 5,6,7-tribromo-4-methyl-1H-benzotriazole (38, IC(50) 0.51 μM) and 5,6,7-tribromo-4-ethyl-1H-benzotriazole (40, IC(50) 0.16 μM). The derivatives with large aromatic or heterocyclic substituents connected to benzimidazole or benzotriazole scaffold appeared to be less potent inhibitors.