Targeting of active sialyltransferase to the plant Golgi apparatus

Glycosyltransferases in the Golgi apparatus synthesize cell wall polysaccharides and elaborate the complex glycans of glycoproteins. To investigate the targeting of this type of enzyme to plant Golgi compartments, we generated transgenic Arabidopsis plants expressing alpha-2,6-sialyltransferase, a glycosyltransferase of the mammalian trans-Golgi cisternae and the trans-Golgi network. Biochemical analysis as well as immunolight and immunoelectron microscopy of these plants indicate that the protein is targeted specifically to the Golgi apparatus. Moreover, the protein is predominantly localized to the cisternae and membranes of the trans side of the organelle. When supplied with the appropriate substrates, the enzyme has significant alpha-2,6-sialyltransferase activity. These results indicate a conservation of glycosyltransferase targeting mechanisms between plant and mammalian cells and also demonstrate that glycosyltransferases can be subcompartmentalized to specific cisternae of the plant Golgi apparatus.     

Development of a high intensity Al beam from SiC targets for accelerated beam experiments at ISAC

An intense beam of ^26gAl has been developed for accelerated beam experiments at TRIUMF’s ISAC facility. Studies of the on-line production of Al radionuclides from thick silicon carbide targets have been performed as part of a program of beam development for astrophysical reaction studies at ISAC. While the release of short-lived Al nuclides from SiC was found to be slow, development of new target material forms and high-power target containers has allowed operation of SiC targets with proton currents of up to 70 μA on target. In addition, operation with the TRIUMF resonant ionization laser ion source (TRILIS) has produced ^26gAl beam intensities of 5.1 × 10¹⁰ s⁻¹.     

The Use of Atomic Force Microscopy in Investigating Particle Caking Mechanisms

Spray‐dried materials are being used increasingly in industries such as food, detergent and pharmaceutical manufacture. Spray‐dried sodium carbonate is an important product that has a great propensity to cake; its moisture‐sorption properties are very different to the crystalline and amorphous species, with a great affinity for atmospheric moisture. This work demonstrates how the noncontact surface analysis of individual particles using atomic force microscopy can highlight the possible mechanisms of unwanted agglomeration. The nondestructive nature of this method allows cycling of localised humidity in situ and repeated scanning of the same particle area. The resulting topography and phase scans showed that humidity cycling caused changes in the distribution of material phases that were not solely dependent on topographical changes.     

Incorporating a Polyethyleneglycol Linker to Enhance the Hydrophilicity of Mitochondria-Targeted Triphenylphosphonium Constructs

The targeting of bioactive molecules and probes to mitochondria can be achieved by coupling to the lipophilic triphenyl phosphonium (TPP) cation, which accumulates several hundred-fold within mitochondria in response to the mitochondrial membrane potential (Δψ_m). Typically, a simple alkane links the TPP to its “cargo”, increasing overall hydrophobicity. As it would be beneficial to enhance the water solubility of mitochondria-targeted compounds we explored the effects of replacing the alkyl linker with a polyethylene glycol (PEG). We found that the use of PEG led to compounds that were readily taken up by isolated mitochondria and by mitochondria inside cells. Within mitochondria the PEG linker greatly decreased adsorption of the TPP constructs to the matrix-facing face of the mitochondrial inner membrane. These findings will allow the distribution of mitochondria-targeted TPP compounds within mitochondria to be fine-tuned.     

Analysis of Powder Caking in Multicomponent Powders Using Atomic Force Microscopy to Examine Particle Properties

Atomic force microscopy has been used to study the surface properties of model spray dried powders. Phase imaging, nanoindentation and force modulation microscopy have differentiated between the different surface material properties of the particles, revealing a regular dispersion of soft, oil rich areas distributed across the particles' surface. Humidity and temperature cycling effects on the caking behavior of the particles have also been investigated, with significant morphology changes and onset of caking found to occur within relatively short periods of time.     

Fibroblast growth factor receptor expression reflects cellular differentiation in human oral squamous carcinoma cell lines

This study examined the expression of fibroblast growth factor receptor 2 (FGFR 2) splice variants, IIIb and IIIc, in normal and malignant human oral keratinocytes and in normal oral fibroblasts by RT-PCR using both exon-specific primers and primers common to both FGFR 2 isoforms. Fibroblasts expressed exclusively FGFR 2/IIIc whilst the normal and malignant keratinocytes co-expressed FGFR 2/IIIb and FGFR 2/IIIc. Well-differentiated keratinocytes expressed proportionally more FGFR 2/IIIb than IIIc whereas the poorly-differentiated cells expressed more FGFR 2/IIIc than IIIb. The normal and malignant keratinocytes, but not fibroblasts, expressed an additional amplification product, which consisted of both IIIb and IIIc of FGFR 2 joined by an extra base pair and with the intronic sequence removed. The results indicate that the expression of FGFR 2 isoforms reflects the degree of cellular differentiation in normal and malignant human oral keratinocytes and that receptor complexes of FGFR 2/IIIb and IIIc may regulate ligand-receptor interactions.     

Differential scanning calorimetry of the epoxy cure reaction

From a review of the literature it is shown that kinetics from programmed heating rate experiments are often in serious disagreement with published isothermal data. In every case, including the present, the dynamic kinetic parameters measured on epoxies are significantly larger than the isothermal. Use of a dynamic rate equation provides good agreement between the two experiments. It is also shown that the Kissinger method is correct when applied to DSC of epoxies.     

When cross-modal spatial attention fails.

There is now convincing evidence that an involuntary shift of spatial attention to a stimulus in one modality can affect the processing of stimuli in other modalities, but inconsistent findings across different paradigms have led to controversy. Such inconsistencies have important implications for theories of cross-modal attention. The authors investigated why orienting attention to a visual event sometimes influences responses to subsequent sounds and why it sometimes fails to do so. They examined visual-cue-on-auditory-target effects in two paradigms--implicit spatial discrimination (ISD) and orthogonal cuing (OC)--that have yielded conflicting findings in the past. Consistent with previous research, visual cues facilitated responses to same-side auditory targets in the ISD paradigm but not in the OC paradigm. Furthermore, in the ISD paradigm, visual cues facilitated responses to auditory targets only when the targets were presented directly at the cued location, not when they appeared above or below the cued location. This pattern of results confirms recent claims that visual cues fail to influence responses to auditory targets in the OC paradigm because the targets fall outside the focus of attention. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Solid Bridge Formation Between Spray-Dried Sodium Carbonate Particles

Moisture migration caking of pharmaceutical excipients in the absence of load is a significant quality and stability issue. This study uses Atomic Force Microscopy (AFM) to examine a solid bridge formed between two 20 µm spray-dried sodium carbonate particles. The bridge is grown by repeatedly exposing the system to 70% RH and 30% RH cycles at 25°C. A comparison is made with the idealized bridge model developed by Tanaka (1978), which was previously verified using crystalline systems. The resulting system was found to be more complex and grew in two stages. The first stage consisted of linear growth to five cycles, followed by a more gradual expansion and the appearance of crystalline structures.     

Effect of processing variables and bulk composition on the surface composition of spray dried powders of a model food system

The surface compositions of food powders created from spray drying solutions containing various ratios of sodium caseinate, maltodextrin and soya oil have been analysed by Electron Spectroscopy for Chemical Analysis. The results show significant enrichment of oil at the surface of particles compared to the bulk phase and, when the non-oil components only are considered, a significant surface enrichment of sodium caseinate also. The degree of surface enrichment of both oil and sodium caseinate was found to increase with decreasing bulk levels of the respective components. Surface enrichment of oil was also affected by processing conditions (emulsion drop size and drying temperature), but surface enrichment of sodium caseinate was relatively insensitive to these. The presence of “pock marks” on the particle surfaces strongly suggests that the surface oil was caused by rupturing of emulsion droplets at the surface as the surrounding matrix contracts and hardens.