Fluorescent T7 display phages obtained by translational frameshift

Lytic phages form a powerful platform for the display of large cDNA libraries and offer the possibility to screen for interactions with almost any substrate. To visualize these interactions directly by fluorescence microscopy, we constructed fluorescent T7 phages by exploiting the flexibility of phages to incorporate modified versions of its capsid protein. By applying translational frameshift sequences, helper plasmids were constructed that expressed a fixed ratio of both wild-type capsid protein (gp10) and capsid protein fused to enhanced yellow fluorescent protein (EYFP). The frameshift sequences were inserted between the 3′ end of the capsid gene and the sequence encoding EYFP. Fluorescent fusion proteins are only formed when the ribosome makes a −1 shift in reading frame during translation. Using standard fluorescence microscopy, we could sensitively monitor the enrichment of specific binders in a cDNA library displayed on fluorescent T7 phages. The perspectives of fluorescent display phages in the fast emerging field of single molecule detection and sorting technologies are discussed

Global analysis of estrogen receptor beta binding to breast cancer cell genome reveals an extensive interplay with estrogen receptor alpha for target gene regulation

Background: Estrogen receptors alpha (ERa) and beta (ERb) are transcription factors (TFs) that mediate estrogen signaling and define the hormone-responsive phenotype of breast cancer (BC). The two receptors can be found co-expressed and play specific, often opposite, roles, with ERb being able to modulate the effects of ERa on gene transcription and cell proliferation. ERb is frequently lost in BC, where its presence generally correlates with a better prognosis of the disease. The identification of the genomic targets of ERb in hormone-responsive BC cells is thus a critical step to elucidate the roles of this receptor in estrogen signaling and tumor cell biology. Results: Expression of full-length ERb in hormone-responsive, ERa-positive MCF-7 cells resulted in a marked reduction in cell proliferation in response to estrogen and marked effects on the cell transcriptome. By ChIP-Seq we identified 9702 ERb and 6024 ERa binding sites in estrogen-stimulated cells, comprising sites occupied by either ERb, ERa or both ER subtypes. A search for TF binding matrices revealed that the majority of the binding sites identified comprise one or more Estrogen Response Element and the remaining show binding matrixes for other TFs known to mediate ER interaction with chromatin by tethering, including AP2, E2F and SP1. Of 921 genes differentially regulated by estrogen in ERb+ vs ERb- cells, 424 showed one or more ERb site within 10 kb. These putative primary ERb target genes control cell proliferation, death, differentiation, motility and adhesion, signal transduction and transcription, key cellular processes that might explain the biological and clinical phenotype of tumors expressing this ER subtype. ERb binding in close proximity of several miRNA genes and in the mitochondrial genome, suggests the possible involvement of this receptor in small non-coding RNA biogenesis and mitochondrial genome functions. Conclusions: Results indicate that the vast majority of the genomic targets of ERb can bind also ERa, suggesting that the overall action of ERb on the genome of hormone-responsive BC cells depends mainly on the relative concentration of both ERs in the cell

The past and the present: a historical outline of Polish research into translations of children\u2019s and young adult literature

The purpose of this contibution is to examine the historical outline of Polish research into translations of children\u2019s and young adult literature, especially between the XXth and the XXIst century. Thanks to several scholar and translator, in the last fifty years these studies have been increased, also in an international way. The contributors report on their research findings and identify the reasons behind the presence or absence of certain translations of children\u2019s and young adult literature in Poland; in fact, the dynamic development of Polish research into translations for young readers interwines different connections, for example, with Western children\u2019s literature, with the censorship or with the field of illustred books for children

The past and the present: a historical outline of Polish research into translations of children\u2019s and young adult literature. Introduction

The purpose of this contibution is to examine the historical outline of Polish research into translations of children\u2019s and young adult literature, especially between the XXth and the XXIst century. Thanks to several scholar and translator, in the last fifty years these studies have been increased, also in an international way. The contributors report on their research findings and identify the reasons behind the presence or absence of certain translations of children\u2019s and young adult literature in Poland; in fact, the dynamic development of Polish research into translations for young readers interwines different connections, for example, with Western children\u2019s literature, with the censorship or with the field of illustred books for children

Effect of composted municipal wastes on concentration of heavy metals in sandy soil in the first year after compost application

Przedstawiono wyniki doświadczenia polowego, którego celem była ocena wpływu zastosowanych dawek kompostowanych odpadów miejskich na właściwości gleby wytworzonej z piasku gliniastego lekkiego, a szczególnie na całkowite zawartości metali ciężkich w glebie. Zastosowano komposty z kompostowni miejskich w Katowicach oraz w Żywcu. Kompost katowicki zawierał znaczne ilości niektórych metali ciężkich: 1850 mg Zn·kg⁻¹, 980 mg Pb·kg⁻¹, 370 mg Cu·kg⁻¹, 12 mg Cd·kg⁻¹ oraz 100 mg Cr·kg⁻¹, w odniesieniu do suchej masy. Zawartości metali ciężkich w kompoście żywieckim były 3-6 razy niższe. Komposty zastosowano do nawożenia gleby w dawkach: 30, 60 i 120 t·kg⁻¹. Obiektami porównawczymi były obiekty bez nawożenia oraz z zastosowaniem nawożenia mineralnego NPK. Przedstawiono wyniki uzyskane w pierwszym roku po zastosowaniu kompostu. Stwierdzono istotny statystycznie wzrost zawartości wszystkich analizowanych metali: Pb, Zn, Cd, Cu, Cr i Ni wraz ze wzrostem dawki kompostu katowickiego oraz brak istotnego wpływu wielkości dawek kompostu żywieckiego na zawartość metali ciężkich w glebie. Podkreślono jednak, że nawet przy zastosowaniu wysokich dawek kompostu katowickiego stopnień zanieczyszczenia gleb metalami ciężkimi, wyznaczony wg wytycznych Instytutu Uprawy Nawożenia i Gleboznawstwa, praktycznie nie zmienia się, i pod względem zawartości poszczególnych metali gleba spełnia kryteria przyjęte dla gleb lekkich niezanieczyszczonych (0 stopień zanieczyszczenia) lub gleb o podwyższonej zawartości metali ciężkich (I stopień), co nie wpływa ograniczająco na produkcję zbóż.A field experiment was carried out to examine the effects of composted municipal wastes applied to sandy soil, on soil properties, in particular on concentrations of heavy metals in soil. Two composts were used, produced in municipal composting plants in Katowice and Żywiec. Compost from Katowice contained high concentrations of heavy metals: 1850 mg Zn·kg⁻¹, 980 mg Pb·kg⁻¹, 370 mg Cu·kg⁻¹, 12 mg Cd·kg⁻¹ and 100 mg Cr·kg⁻¹, whereas their concentrations in compost from Żywiec were by 3-6 fold lower. Composting ratios were: 30, 60 and 120 t per ha. Fields without fertilization and fertilized with mineral NPK were used as control objects. Results presented in the paper were obtained after first year of experiment. Concentrations of all examined heavy metals (Pb, Zn, Cd, Cu, Cr, Ni) increased significantly with increasing composting rates in case of compost from Katowice, while the compost from Żywiec did not cause any significant changes in concentrations of heavy metals in soils. It was stressed that the application of compost from Katowice did not cause any vital changes in a degree of soil pollution with heavy metals, as determined according to The Institute of Soil Science and Plant Cultivation (IUNG) guidelines, and even with a high rate of amendment (120 t per ha) the soil concentrations of metals fulfilled the criteria for unpolluted soils (0 degree of pollution) or soils with enhanced concentrations of heavy metals (I degree of pollution), without any limitations for crop production