Glycosylation pattern of brush border-associated glycoproteins in enterocyte-like cells: involvement of complex-type N-glycans in apical trafficking

We have previously reported that galectin-4, a tandem repeat-type galectin, regulates the raft-dependent delivery of glycoproteins to the apical brush border membrane of enterocyte-like HT-29 cells. N-Acetyllactosamine-containing glycans, known as galectin ligands, were found enriched in detergent-resistant membranes. Here, we analyzed the potential contribution of N-and/ or O-glycans in this mechanism. Structural studies were carried out on the brush border membrane-enriched fraction using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) and nano-ESI-QTOF-MS/MS. The pattern of N-glycans was very heterogeneous, with the presence of high mannose- and hybrid-type glycans as well as a multitude of complex-type glycans. In contrast, the pattern of O-glycans was very simple with the presence of two major core type 1 O-glycans, sialylated and bisialylated T-antigen structures {[}Neu5Ac alpha 2-3Gal beta 1-3GalNAc-ol and Neu5Ac alpha 2-3Gal beta 1 -3(Neu5Ac alpha 2-6)GalNAc-ol]. Thus, N-glycans rather than O-glycans contain the N-acetyllactosamine recognition signals for the lipid raft-based galectin-4-dependent apical delivery. In the presence of 1-deoxymannojirimycin, a drug which inhibits the generation of hybrid-type or complex type N-glycans, the extensively O-glycosylated mucin-like MUC1 glycoprotein was not delivered to the apical brush border but accumulated inside the cells. Altogether, our data demonstrate the crucial role of complex N-glycans in the galectin-4-dependent delivery of glycoproteins to the apical brush border membrane of enterocytic HT-29 cells

Gedanken experiments with Casimir forces, vacuum energy, and gravity

Gedanken experiments are used to explore properties of quantum vacuum energy that are currently challenging to explore experimentally. A constant lateral Casimir force is predicted to exist between two overlapping finite parallel plates at 0 K, otherwise it would be possible to extract an arbitrary amount of energy from the quantum vacuum. A rigid unpowered object cannot be accelerated by the quantum vacuum because of the translational symmetry of space. By considering systems in which vacuum energy and other forms of energy are exchanged, we demonstrate that a change {\Delta}E in vacuum energy, whether positive or negative with respect to the free field, corresponds to an equivalent inertial mass and equivalent gravitational mass {\Delta}M={\Delta}E/c^2. We consider the possibility of a gravitational shield, and show that, if it exists, the energy to operate it would have to cancel the net energy extracted from the gravitational field, otherwise we could extract an arbitrary amount of energy from the field.Comment: 24 pages, 3 figure

Imagination and Film

This chapter addresses the application of contemporary theories of the imagination—largely drawn from cognitive psychology—to our understanding of film. Topics include the role of the imagination in our learning what facts hold within a fictional film, including what characters’ motivations, beliefs, and feelings are; how our perceptual experience of a film explains our imaginative visualizing of its contents; how fictional scenarios in films generate certain affective and evaluative responses; and how such responses compare to those we have toward analogous circumstances in real life

Glycocalyx Restricts Adenoviral Vector Access to Apical Receptors Expressed on Respiratory Epithelium In Vitro and In Vivo: Role for Tethered Mucins as Barriers to Lumenal Infection

Inefficient adenoviral vector (AdV)-mediated gene transfer to the ciliated respiratory epithelium has hindered gene transfer strategies for the treatment of cystic fibrosis lung disease. In part, the inefficiency is due to an absence of the coxsackie B and adenovirus type 2 and 5 receptor (CAR) from the apical membranes of polarized epithelia. In this study, using an in vitro model of human ciliated airway epithelium, we show that providing a glycosylphosphatidylinositol (GPI)-linked AdV receptor (GPI-CAR) at the apical surface did not significantly improve AdV gene transfer efficiency because the lumenal surface glycocalyx limited the access of AdV to apical GPI-CAR. The highly glycosylated tethered mucins were considered to be significant glycocalyx components that restricted AdV access because proteolytic digestion and inhibitors of O-linked glycosylation enhanced AdV gene transfer. To determine whether these in vitro observations are relevant to the in vivo situation, we generated transgenic mice expressing GPI-CAR at the surface of the airway epithelium, crossbred these mice with mice that were genetically devoid of tethered mucin type 1 (Muc1), and tested the efficiency of gene transfer to murine airways expressing apical GPI-human CAR (GPI-hCAR) in the presence and absence of Muc1. We determined that AdV gene transfer to the murine airway epithelium was inefficient even in GPI-hCAR transgenic mice but that the gene transfer efficiency improved in the absence of Muc1. However, the inability to achieve a high gene transfer efficiency, even in mice with a deletion of Muc1, suggested that other glycocalyx components, possibly other tethered mucin types, also provide a significant barrier to AdV interacting with the airway lumenal surface

Pretense and Imagination

Issues of pretense and imagination are of central interest to philosophers, psychologists, and researchers in allied fields. In this entry, we provide a roadmap of some of the central themes around which discussion has been focused. We begin with an overview of pretense, imagination, and the relationship between them. We then shift our attention to the four specific topics where the disciplines' research programs have intersected or where additional interactions could prove mutually beneficial: the psychological underpinnings of performing pretense and of recognizing pretense, the cognitive capacities involved in imaginative engagement with fictions, and the real-world impact of make-believe. In the final section, we discuss more briefly a number of other mental activities that arguably involve imagining, including counterfactual reasoning, delusions, and dreaming

Verbal Reports and "Real' Reasons" : Confabulation and Conflation

This document is the Accepted Manuscript version of a published work that appeared in final form in Ethical Theory and Moral Practice after peer review and technical editing by the publisher. Constantine Sandis, ‘Verbal Reports and “Real” Reasons: Confabulation and Conflation’, Ethical Theory and Moral Practice, Vol. 18(2): 267-280, first published online 18 March 2015. The final publication is available at Springer via http://dx.doi.org/10.1007/s10677-015-9576-6 © Springer Science+Business Media Dordrecht 2015This paper examines the relation between the various forces which underlie human action and verbal reports about our reasons for acting as we did. I maintain that much of the psychological literature on confabulations rests on a dangerous conflation of the reasons for which people act with a variety of distinct motivational factors. In particular, I argue that subjects frequently give correct answers to questions about the considerations they acted upon while remaining largely unaware of why they take themselves to have such reasons to act. Pari passu, experimental psychologists are wrong to maintain that they have shown our everyday reason talk to be systematically confused. This is significant because our everyday reason-ascriptions affect characterizations of action (in terms of intention, knowledge, foresight, etc.) that are morally and legally relevant. I conclude, more positively, that far from rendering empirical research on confabulations invalid, my account helps to reveal its true insights into human nature.Peer reviewe

MUC1 alters oncogenic events and transcription in human breast cancer cells

INTRODUCTION: MUC1 is an oncoprotein whose overexpression correlates with aggressiveness of tumors and poor survival of cancer patients. Many of the oncogenic effects of MUC1 are believed to occur through interaction of its cytoplasmic tail with signaling molecules. As expected for a protein with oncogenic functions, MUC1 is linked to regulation of proliferation, apoptosis, invasion, and transcription. METHODS: To clarify the role of MUC1 in cancer, we transfected two breast cancer cell lines (MDA-MB-468 and BT-20) with small interfering (si)RNA directed against MUC1 and analyzed transcriptional responses and oncogenic events (proliferation, apoptosis and invasion). RESULTS: Transcription of several genes was altered after transfection of MUC1 siRNA, including decreased MAP2K1 (MEK1), JUN, PDGFA, CDC25A, VEGF and ITGAV (integrin α(v)), and increased TNF, RAF1, and MMP2. Additional changes were seen at the protein level, such as increased expression of c-Myc, heightened phosphorylation of AKT, and decreased activation of MEK1/2 and ERK1/2. These were correlated with cellular events, as MUC1 siRNA in the MDA-MB-468 line decreased proliferation and invasion, and increased stress-induced apoptosis. Intriguingly, BT-20 cells displayed similar levels of apoptosis regardless of siRNA, and actually increased proliferation after MUC1 siRNA. CONCLUSION: These results further the growing knowledge of the role of MUC1 in transcription, and suggest that the regulation of MUC1 in breast cancer may be more complex than previously appreciated. The differences between these two cell lines emphasize the importance of understanding the context of cell-specific signaling events when analyzing the oncogenic functions of MUC1, and caution against generalizing the results of individual cell lines without adequate confirmation in intact biological systems