Inhibition of human lung cancer cell proliferation and survival by wine

Compounds of plant origin and food components have attracted scientific attention for use as agents for cancer prevention and treatment. Wine contains polyphenols that were shown to have anti-cancer and other health benefits. The survival pathways of Akt and extracellular signal-regulated kinase (Erk), and the tumor suppressor p53 are key modulators of cancer cell growth and survival. In this study, we examined the effects of wine on proliferation and survival of human Non-small cell lung cancer (NSCLC) cells and its effects on signaling events.Compounds of plant origin and food components have attracted scientific attention for use as agents for cancer prevention and treatment. Wine contains polyphenols that were shown to have anti-cancer and other health benefits. The survival pathways of Akt and extracellular signal-regulated kinase (Erk), and the tumor suppressor p53 are key modulators of cancer cell growth and survival. In this study, we examined the effects of wine on proliferation and survival of human Non-small cell lung cancer (NSCLC) cells and its effects on signaling events

Mapping body-building potential

Experiments in mice shed new light on an elusive population of embryonic cells called neuromesodermal progenitors

Major transcriptome re-organisation and abrupt changes in signalling, cell cycle and chromatin regulation at neural differentiation <em>in vivo</em>

Here, we exploit the spatial separation of temporal events of neural differentiation in the elongating chick body axis to provide the first analysis of transcriptome change in progressively more differentiated neural cell populations in vivo. Microarray data, validated against direct RNA sequencing, identified: (1) a gene cohort characteristic of the multi-potent stem zone epiblast, which contains neuro-mesodermal progenitors that progressively generate the spinal cord; (2) a major transcriptome reorganisation as cells then adopt a neural fate; and (3) increasing diversity as neural patterning and neuron production begin. Focussing on the transition from multi-potent to neural state cells, we capture changes in major signalling pathways, uncover novel Wnt and Notch signalling dynamics, and implicate new pathways (mevalonate pathway/steroid biogenesis and TGF beta). This analysis further predicts changes in cellular processes, cell cycle, RNA-processing and protein turnover as cells acquire neural fate. We show that these changes are conserved across species and provide biological evidence for reduced proteasome efficiency and a novel lengthening of S phase. This latter step may provide time for epigenetic events to mediate large-scale transcriptome re-organisation; consistent with this, we uncover simultaneous downregulation of major chromatin modifiers as the neural programme is established. We further demonstrate that transcription of one such gene, HDAC1, is dependent on FGF signalling, making a novel link between signals that control neural differentiation and transcription of a core regulator of chromatin organisation. Our work implicates new signalling pathways and dynamics, cellular processes and epigenetic modifiers in neural differentiation in vivo, identifying multiple new potential cellular and molecular mechanisms that direct differentiation

Lineage tracing of axial progenitors using Nkx1-2CreERT2 mice defines their trunk and tail contributions

The vertebrate body forms by continuous generation of new tissue from progenitors at the posterior end of the embryo. The study of these axial progenitors has proved challenging in vivo largely due to the lack of unique molecular markers to identify them. Here, we elucidate the expression pattern of the transcription factor Nkx1-2 in the mouse embryo and show that it identifies axial progenitors throughout body axis elongation, including neuromesodermal progenitors and early neural and mesodermal progenitors. We create a tamoxifen-inducible Nkx1-2CreERT2 37 transgenic mouse and exploit the conditional nature of this line to uncover the lineage contributions of Nkx1-2-expressing cells at specific stages. We show that early Nkx1-2-expressing epiblast cells contribute to all three germ layers, mostly neuroectoderm and mesoderm, excluding notochord. Our data are consistent with the presence of some self-renewing axial progenitors that continue to generate neural and mesoderm tissues from the tail bud. This study identifies Nkx1-2 expressing cells as the source of most trunk and tail tissues in the mouse and provides a useful tool to genetically label and manipulate axial progenitors in vivo

Assessing the efficiency of novel gene trap vectors in murine embryonic stem cells

Gene trapping is a random insertional mutagenesis strategy that aims to identify novel genes and analyse their function. It usually involves the introduction into embryonic stem (ES) cells of promoterless reporter/selector gene constructs whose expression can be activated only after integration downstream of a gene's regulatory elements. Gene trap insertions result in production of fusion transcripts consisting of the reporter and endogenous sequences and the mutated genes can be readily identified using PCR-based methods such as RACE. Furthermore the biological consequences of the integration event can be assessed after germ-line transmission. One limitation of conventional gene trapping is that it can only target genes expressed in ES cells since selection of insertional events relies on the endogenous promoter's activity to drive expression of the selectable marker and to circumvent this problem a new class of gene trap vectors called poly(A) trap vectors was developed. These constructs contain a 3' selectable marker whose expression is driven by a constitutively active internal promoter relaxing the requirement for endogenous gene expression. The selectable marker lacks a polyA signal but incorporates a splice donor (SD) signal so only integrations upstream of an endogenous gene's splice acceptor (SA) and polyA sequences can be selected thus eliminating intergenic background insertions. However, it has been recently demonstrated that poly(A) trap vectors are biased towards integrations into the 3'most-intron of their target genes due to the action of an mRNA-surveillance mechanism called nonsense-mediated mRNA decay (NMD).The aim of the study presented here was to assess the efficiency of a series of gene trap vectors that incorporate two novel features in their design: (i) the presence of an ATG-less, 5' triple fusion between egfp, P-galactosidase and neomycin/hygromycin resistance genes to function as a reporter/selector of the trapped gene's expression state and (ii) a 3' poly(A) trap cassette that contains the previously uncharacterized rabbit (3-globin exon 2/intron 2 SD junction and an AU-rich element (ARE) derived from the human GM-CSF gene. Our results provide evidence that the triple fusion functions properly and can be potentially used as a reporter of trapped locus activity. We also show that the presence of the ARE appears to improve the performance of the rabbit (3-globin SD sequence in the context of poly(A) trapping. More importantly, preliminary data suggest that our vectors may be resistant to NMD and thus potentially unbiased in their insertional preference

A human iPSC line capable of differentiating into functional macrophages expressing ZsGreen: a tool for the study and in vivo tracking of therapeutic cells

We describe the production of a human induced pluripotent stem cell (iPSC) line, SFCi55-ZsGr, that has been engineered to express the fluorescent reporter gene, ZsGreen, in a constitutive manner. The CAG-driven ZsGreen expression cassette was inserted into the AAVS1 locus and a high level of expression was observed in undifferentiated iPSCs and in cell lineages derived from all three germ layers including haematopoietic cells, hepatocytes and neurons. We demonstrate efficient production of terminally differentiated macrophages from the SFCi55-ZsGreen iPSC line and show that they are indistinguishable from those generated from their parental SFCi55 iPSC line in terms of gene expression, cell surface marker expression and phagocytic activity. The high level of ZsGreen expression had no effect on the ability of macrophages to be activated to an M(LPS + IFNγ), M(IL10) or M(IL4) phenotype nor on their plasticity, assessed by their ability to switch from one phenotype to another. Thus, targeting of the AAVS1 locus in iPSCs allows for the production of fully functional, fluorescently tagged human macrophages that can be used for in vivo tracking in disease models. The strategy also provides a platform for the introduction of factors that are predicted to modulate and/or stabilize macrophage function. This article is part of the theme issue ‘Designer human tissue: coming to a lab near you’

Understanding axial progenitor biology in vivo and in vitro

The generation of the components that make up the embryonic body axis, such as the spinal cord and vertebral column, takes place in an anterior-to-posterior (head-to-tail) direction. This process is driven by the coordinated production of various cell types from a pool of posteriorly-located axial progenitors. Here, we review the key features of this process and the biology of axial progenitors, including neuromesodermal progenitors, the common precursors of the spinal cord and trunk musculature. We discuss recent developments in the in vitro production of axial progenitors and their potential implications in disease modelling and regenerative medicine

MINERALOGICAL AND SPECTROSCOPIC STUDY OF NESQUEHONITE SYNTHESIZED BY REACTION OF GASEOUS CO2 WITH MG CHLORIDE SOLUTION

Στην παρούσα εργασία πραγματοποιήθηκε η σύνθεση νεσκεχονίτη, ενός ένυδρου ανθρακικού ορυκτού, υπό χαμηλές συνθήκες πίεσης με αντίδραση CO2 σε διάλυμα χλωριούχου μαγνησίου. Ο νεσκεχονίτης μπορεί να αξιοποιηθεί ως πρώτη ύλη σε δομικά υλικά και επιπλέον στην διαχείριση υγρών αποβλήτων. Ο νεσκεχονίτης μελετήθηκε με περιθλασιομετρία ακτίνων-Χ, υπέρυθρη φασματοσκοπία (FT-IR) και φασματοσκοπία Raman, διοφθάλμιο στερεοσκόπιο, Ηλεκτρονικό Μικροσκόπιο Σάρωσης και Ηλεκτρονικό Μικροσκόπιο Διερχόμενης Δέσμης Ηλεκτρονίων. Ο παραγόμενος νεσκεχονίτης αναπτύσσει επιμήκεις διαφανείς έως ημιδιαφανείς βελονοειδείς κρυστάλλους με υαλώδη λάμψη. Η υπέρυθρη φασματοσκοπία (FT-ΙR) και η φασματοσκοπία Raman υπέδειξαν την παρουσία ΟΗ- και HCO3 - στην κρυσταλλική δομή του νεσκεχονίτη. Η διαδικασία σύνθεσης που περιγράφεται στην παρούσα εργασία μπορεί να χρησιμοποιηθεί στην διαδικασία της ορυκτοποίησης για μόνιμη αποθήκευση των εκπομπών CO2Nesquehonite, a hydrous carbonate with promising uses such as building raw material and treatment of wastewaters, was synthesized under low pressure conditions by reaction of gaseous CO2 with Mg chloride solution and it was studied by means of X-Ray Diffraction, optical and scanning/transmission electron microscopy, and FTIR and Raman spectroscopic methods. Synthesized nesquehonite forms elongated fibers, exhibiting transparent to translucent diaphaneity and vitreous luster. It is characterized by high crystallinity. IR and Raman spectroscopy indicated the presence of OHand HCO3 - in the crystal structure of nesquehonite. The nesquehonite synthesis described herein constitutes a potential permanent storage of CO2 emissions

miR-23b regulates cytoskeletal remodeling, motility and metastasis by directly targeting multiple transcripts

Uncontrolled cell proliferation and cytoskeletal remodeling are responsible for tumor development and ultimately metastasis. A number of studies have implicated microRNAs in the regulation of cancer cell invasion and migration. Here, we show that miR-23b regulates focal adhesion, cell spreading, cell-cell junctions and the formation of lamellipodia in breast cancer (BC), implicating a central role for it in cytoskeletal dynamics. Inhibition of miR-23b, using a specific sponge construct, leads to an increase of cell migration and metastatic spread in vivo, indicating it as a metastatic suppressor microRNA. Clinically, low miR-23b expression correlates with the development of metastases in BC patients. Mechanistically, miR-23b is able to directly inhibit a number of genes implicated in cytoskeletal remodeling in BC cells. Through intracellular signal transduction, growth factors activate the transcription factor AP-1, and we show that this in turn reduces miR-23b levels by direct binding to its promoter, releasing the pro-invasive genes from translational inhibition. In aggregate, miR-23b expression invokes a sophisticated interaction network that co-ordinates a wide range of cellular responses required to alter the cytoskeleton during cancer cell motility