P-selectin mediates metastatic progression through binding to sulfatides on tumor cells

Hematogenous carcinoma metastasis is associated with tumor cell emboli formation, which is now known to be facilitated by selectins. P-selectin-mediated interactions of platelets with cancer cells are based mostly on mucin- and glycosaminoglycan-type selectin ligands. We previously showed that mouse colon carcinoma cells (MC-38) carry P-selectin ligands of nonmucin origin, which were not identified. Here we show that P-selectin ligands recognized on MC-38 cells are sulfated glycolipids, thereby facilitating experimental metastasis in a syngeneic mouse model. Metabolic inhibition of sulfation by incubation of cells with sodium chlorate almost completely abrogated P-selectin binding. Metabolic labeling of MC-38 cells with (35)S sulfate revealed only a single band as detected by high-performance thin layer chromatography analysis of a total lipid extract. Matrix-assisted laser desorption/ionization tandem time-of-flight/time-of-flight analysis (MALDI-TOF-TOF) analysis of the purified sulfate-containing lipid fraction identified the selectin ligand to be a sulfated galactosylceramide SM4 (HSO(3)-3Galbeta-1Cer). Modulation of glycolipid biosynthesis in MC-38 cells altered P-selectin binding, thereby confirming sulfoglycolipids to be major P-selectin ligands. In addition, P-selectin was also found to recognize lactosylceramide sulfate SM3 (HSO(3)-3Galbeta-4Glcbeta-1Cer) and gangliotriaosylceramide sulfate SM2 [GalNAcbeta-4(HSO(3)-3)Galbeta-4Glcbeta-1Cer] in human hepatoma cells. Finally, the enzymatic removal of sulfation from the cell surface of MC-38 cells resulted in decreased P-selectin binding and led to attenuation of metastasis. Thus, SM4 sulfatide serves as a native ligand for P-selectin contributing to cell-cell interactions and to facilitation of metastasi

Production, purification and crystallization of a trans-sialidase from Trypanosoma vivax

Sialidases and trans-sialidases play important roles in the life cycles of various microorganisms. These enzymes can serve nutritional purposes, act as virulence factors or mediate cellular interactions (cell evasion and invasion). In the case of the protozoan parasite Trypanosoma vivax, trans-sialidase activity has been suggested to be involved in infection-associated anaemia, which is the major pathology in the disease nagana. The physiological role of trypanosomal trans-sialidases in host-parasite interaction as well as their structures remain obscure. Here, the production, purification and crystallization of a recombinant version of T. vivaxtrans-sialidase 1 (rTvTS1) are described. The obtained rTvTS1 crystals diffracted to a resolution of 2.5 angstrom and belonged to the orthorhombic space group P2(1)2(1)2(1), with unit-cell parameters a = 57.3, b = 78.4, c = 209.0 angstrom

3D Microstructured Carbon Nanotube Electrodes for Trapping and Recording Electrogenic Cells

Electrogenic cells such as cardiomyocytes and neurons rely mainly on electrical signals for intercellular communication. Microelectrode arrays (MEAs) have been developed for long-term recording of cell signals and stimulation of electrogenic cells under low-cell-stress conditions, providing new insights in the behavior of electrogenic cells and the operation of the brain. To date, MEAs are relying on flat or needle-shaped electrode surfaces, mainly due to limitations in the lithographic processes. This paper relies on a previously reported elasto-capillary aggregation process to create 3D carbon nanotube (CNT) MEAs. This study shows that CNTs aggregate in well-shaped structures of similar size as cardiomyocytes are particularly interesting for MEA applications. This is because i) CNT microwells of the right diameter preferentially trap individual cardiomyocytes, which facilitates single cell recording without the need for clamping cells or signal deconvolution, and ii) once the cells are trapped inside of the CNT wells, this 3D CNT structure is used as an electrode surrounding the cell, which increases the cell-electrode contact area. As a result, this study finds that the recorded output voltages increase significantly (more than 200%). This fabrication process paves the way for future study of complex interactions between electrogenic cells and 3D recording electrodes.This work was supported by the Research Foundation—Flanders (FWO, Belgium) under Project No. 11S1214N. Michael De Volder was supported by the ERC Starting Grant (337739)—HIENA and the Marie Curie Grant CANA (618250). Davor Copic was supported by the Marie Curie Grant EmuCam (660351)

Influence of Temperature on Post-Breakage Behaviour of Laminated Glass Beams : Experimental Approach

The assessment of the post-breakage performances of laminated glass elementsused in construction need to take into account the sensitivity to the temperature ofthe mechanical behaviour and properties of the product, in particular of theinterlayer material. A general problem statement and an overview of differentexperimental approaches are firstly presented. Then results of specific orientationtests on pre-cracked laminated glass beams with a stiff interlayer of DuPont carriedat three different temperatures (23, 45 and 60°C) are presented and commented. Acomparison of the mechanical behaviour at the different temperatures is done,aiming to give a comprehensive order of magnitude of the sensitivity totemperature of the post-breakage behaviour observed during the tests

Wettability and osteoblast cell response modulation through UV laser processing of nylon 6,6

With an ageing population the demand for cheap, efficient implants is ever increasing. Laser surface treatment offers a unique means of varying biomimetic properties to determine generic parameters to predict cell responses. This paper details how a KrF excimer laser can be employed for both laser-induced patterning and whole area irradiative processing to modulate the wettability characteristics and osteoblast cell response following 24 hour and 4 day incubation. Through white light interferometry (WLI) it was found that the surface roughness had considerably increased by up to 1.5 µm for the laser-induced patterned samples and remained somewhat constant at around 0.1 µm for the whole area irradiative processed samples. A sessile drop device determined that the wettability characteristics differed between the surface treatments. For the patterned samples the contact angle, θ, increased by up to 25° which can be attributed to a mixed-state wetting regime. For the whole area irradiative processed samples θ decreased owed to an increase in polar component, γP. For all samples θ was a decreasing function of the surface energy. The laser whole area irradiative processed samples gave rise to a distinct correlative trend between the cell response, θ and γP. However, no strong relationship was determined for the laser-induced patterned samples due to the mixed-state wetting regime. As a result, owed to the relationships and evidence of cell differentiation one can deduce that laser whole area irradiative processing is an attractive technology for employment within regenerative medicine to meet the demands of an ageing population

A CMOS Time to Digital Converter IC with 2 Level Analog CAM

A time to charge converter IC with an analog memory unit (TCCAMU) has been designed and fabricated in HP\u27s CMOS 1.2-µm n-well process. The TCCAMU is an event driven system designed for front end data acquisition in high energy physics experiments. The chip includes a time to charge converter, analog Level 1 and Level 2 associative memories for input pipelining and data filtering, and an A/D converter. The intervals measured and digitized range from 8-24 ns. Testing of the fabricated chip resulted in an LSB width of 107 ps, a typical differential nonlinearity of \u3c 35 ps, and a typical integral nonlinearity of \u3c 200 ps. The average power dissipation is 8.28 mW per channel. By counting the reference clock, a time resolution of 107 ps over ~ 1 s range could be realized