GOLPH3 Regulates EGFR in T98G Glioblastoma Cells by Modulating Its Glycosylation and Ubiquitylation

Protein trafficking is altered when normal cells acquire a tumor phenotype. A key subcellular compartment in regulating protein trafficking is the Golgi apparatus, but its role in carcinogenesis is still not well defined. Golgi phosphoprotein 3 (GOLPH3), a peripheral membrane protein mostly localized at the trans-Golgi network, is overexpressed in several tumor types including glioblastoma multiforme (GBM), the most lethal primary brain tumor. Moreover, GOLPH3 is currently considered an oncoprotein, however its precise function in GBM is not fully understood. Here, we analyzed in T98G cells of GBM, which express high levels of epidermal growth factor receptor (EGFR), the effect of stable RNAi-mediated knockdown of GOLPH3. We found that silencing GOLPH3 caused a significant reduction in the proliferation of T98G cells and an unexpected increase in total EGFR levels, even at the cell surface, which was however less prone to ligand-induced autophosphorylation. Furthermore, silencing GOLPH3 decreased EGFR sialylation and fucosylation, which correlated with delayed ligand-induced EGFR downregulation and its accumulation at endo-lysosomal compartments. Finally, we found that EGF failed at promoting EGFR ubiquitylation when the levels of GOLPH3 were reduced. Altogether, our results show that GOLPH3 in T98G cells regulates the endocytic trafficking and activation of EGFR likely by affecting its extent of glycosylation and ubiquitylation.     

User interfaces for communication bridges across the digital divide

Connecting people across the “digital divide” is as much a social effort as a technological one. We are developing a community-centred approach to learn how interaction techniques can compensate for poor communication across the digital divide. We have incorporated the lessons learned regarding social intelligence design in an abstraction and in a device called the SoftBridge. The SoftBridge allows communication to flow from endpoints through adapters, getting converted if necessary, and out to destination endpoints. Field trials are underway with two communities in South Africa: disadvantaged Deaf users and an isolated rural community. Initial lessons learned show that we have to design user interfaces that allow users to understand and cope with delay. We also learned that social concerns are often more important than the technical issues in designing such systems.     

Analysis of Combined Systems of Two Endoreversible Engines

A single endoreversible engine can operate as a cooler, a true heat engine, a heat pump, or a refrigerator. We investigate how many different modes of operation a combined system of two endoreversible engines may display. Special attention is paid to the independent combined system which neither consumes nor supplies power.     

GD3/proteosome vaccines induce consistent IgM antibodies against the ganglioside GD3

The gangliosides of melanoma and other tumours of neuroectodermal origin are suitable targets for immune intervention with tumour vaccines. The optimal vaccines in current use contain ganglioside plus bacillus Calmette-Guérin and induce considerable morbidity. We have screened a variety of new adjuvants in the mouse, and describe one antigen-delivery system, proteosomes, which is especially effective. Highly hydrophobic Neisserial outer membrane proteins (OMP) form multimolecular liposome-like vesicular structures termed proteosomes which can readily incorporate amphiphilic molecules such as GD3 ganglioside. The optimal GD3/proteosome vaccine formulation for induction of GD3 antibodies in the mouse is determined. Interestingly, the use of potent immunological adjuvants in addition to proteosomes augments the IgM and IgG antibody titres against OMP in these vaccines but GD3 antibody titres are unaffected. The application of proteosomes to enhance the immune response to GD3 extends the concept of the proteosome immunopotentiating system from lipopeptides to amphipathic carbohydrate epitopes such as cell-surface gangliosides. The demonstrated safety of meningococcal OMP in humans and the data in mice presented here suggest that proteosome vaccines have potential for augmenting the immunogenicity of amphipathic tumour antigens in humans.     

An alcohol expectancy-challenge prevention program for at risk college women.

Alcohol expectancy-challenge programs are effective in changing expectancies and reducing drinking in college men (J. Darker & M. S. Goldman. 1993. 1998); however, recent evidence suggests this protocol might not be effective for women (M. E. Dunn, C. Lau, & I. Y. Cruz, 2000). This study was designed to reevaluate the effectiveness of a social/sexual expectancy-challenge intervention in college women reporting moderate to heavy alcohol use. Forty-six undergraduate women were randomly assigned to the prevention or control condition. Participants reported alcohol expectancies at pre- and posttest and monitored their drinking patterns daily for 6 weeks. The program was effective in changing some expectancies but did not reduce drinking. This further confirms differences in the mechanisms by which expectancy-challenge programs function for men and women. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Neuropeptide Y and luteinizing hormone releasing hormone synergize to stimulate the development of cellular follicle-stimulating hormone in the hamster adenohypophysis

Luteinizing hormone releasing hormone (LHRH) stimulates the development of cellular FSH immunoreactivity in the perinatal hamster adenohypophysis. Because neuropeptide Y (NPY) can act directly on rat adenohypophysial cells to stimulate FSH and LH release and potentiate the stimulatory effect of LHRH on FSH and LH release, we investigated the effects of NPY alone and in combination with a low, ineffective dose of LHRH on inducing cellular FSH immunoreactivity in the neonatal hamster adenohypophysis. Neonatal female pituitary glands were grafted beneath the right renal capsules of hypophysectomized-ovariectomized adult hamster hosts with a catheter implanted in the external jugular vein. After treatment, hosts were decapitated and graft tissue was stained for FSH and LH immunoreactivity. The mean percentage of adenohypophysial cells that stained for FSH was low (2.8%) in grafts in hosts infused continuously with heparinized saline vehicle for 7 days. In other hosts, peptides were pulsed through the catheter every 12 h for 7 days. The mean percentage of FSH cells also was low after pulsing 6 ng LHRH or 2 micrograms NPY but increased substantially when the two peptides were pulsed simultaneously. No differences in the mean percentage of LH cells existed between any of the groups. The results demonstrate that NPY and LHRH can synergize to induce cellular FSH immunoreactivity in the neonatal female hamster.     

Two new decoding algorithms for Reed-Solomon codes

The subject of decoding Reed-Solomon codes is considered. By reformulating the Berlekamp and Welch key equation and introducing new versions of this key equation, two new decoding algorithms for Reed-Solomon codes will be presented. The two new decoding algorithms are significant for three reasons. Firstly the new equations and algorithms represent a novel approach to the extensively researched problem of decoding Reed-Solomon codes. Secondly the algorithms have algorithmic and implementation complexity comparable to existing decoding algorithms, and as such present a viable solution for decoding Reed-Solomon codes. Thirdly the new ideas presented suggest a direction for future research. The first algorithm uses the extended Euclidean algorithm and is very efficient for a systolic VLSI implementation. The second decoding algorithm presented is similar in nature to the original decoding algorithm of Peterson except that the syndromes do not need to be computed and the remainders are used directly. It has a regular structure and will be efficient for implementation only for correcting a small number of errors. A systolic design for computing the Lagrange interpolation of a polynomial, which is needed for the first decoding algorithm, is also presented.This research was supported by a grant from the Canadian Institute for Telecommunications Research under the NCE program of the Government of Canada