Erythrocyte ghost cell-alkaline phosphatase: construction and characterization of a vesicular system for use in biomineralization studies

Alkaline phosphatase is required for the mineralization of bone and cartilage. This enzyme is localized in the matrix vesicle, which plays a role key in calcifying cartilage. In this paper we standardize a method to construction a resealed ghost cell-alkaline phosphatase system to mimic matrix vesicles and examine the kinetic behavior of the incorporated enzyme. Polidocanol-solubilized alkaline phosphatase, free of detergent, was incorporated into resealed ghost cells. This process was time-dependent and practically 50% of the enzyme was incorporated into the vesicles in 40 h of incubation, at 25 degreesC. Alkaline phosphatase-ghost cell systems were relatively homogeneous with diameters of about 300 nm and were more stable when stored at -20 degreesC.Alkaline phosphatase was completely released from the resealed ghost cell-system using only phospholipase C. These experiments confirm that the interaction between alkaline phosphatase and the lipid bilayer of resealed ghost cell is exclusively via glycosylphosphatidylinositol (GPI) anchor of the enzyme.An important point shown is that an enzyme bound to resealed ghost cell does not lose the ability to hydrolyze ATP, pyrophosphate and p-nitrophenyl phosphate (PNPP), but the presence of a ghost membrane, as a support of the enzyme, affects its kinetic properties. Moreover, calcium ions stimulate and phosphate ions inhibit the PNPPase activity of alkaline phosphatase present in resealed ghost cells. (C) 2002 Elsevier B.V. B.V. All rights reserved.

Mammalian brain plasma membranes: Isolation, enzymatic, and chemical characterization

Two variants of a simplified procedure for the isolation of plasma membrane fractions from monkey and rat brains, are described. The preparations show marked enrichments in the marker enzymes, (Na+-K+) adenosine triphosphatase, acetylcholinesterase, 5′-nucleotidase and adenylate cyclase. Lipid analysis and a protein electrophoretic pattern are presented. An enzymatic check has been made to assess for contamination by other cellular organelles. The amino acid composition of brain membrane proteins show a resemblance to the reported composition of erythrocyte ghost proteins but differ from myelin proteins.

CHARACTERISTICS OF ZINC-BINDING TO HUMAN RED-BLOOD-CELL MEMBRANES

The objective of the present study was to standardize the analysis of zinc binding on human red blood cell (RBC) membranes in 20 normal adults. The displacement studies revealed that at the maximal stable zinc concentration tested (600 muM), 57% (mean) of the bound Zn-65 was displaced and to displace half maximal Zn-65, the stable zinc concentration was 300 muM. Scatchard plots revealed two classes of binding sites for zinc on RBC membranes: one with higher affinity, Kd = 1.20 x 10(-5) M (site I), and the other with lower affinity, Kd = 2.77 x 10(-4) M (site II). Binding sites occupancy was 97% means and 58.5% means for sites I and 11, respectively. The displacement was affected by temperature, membrane protein concentration, freezing, thawing, and dialysis. Other metal cations, including Co++, Fe++, and Mn++, had very little effect on Zn-65 displacement, in contrast copper displaced Zn-65 from its binding sites on RBC membranes. Zinc binding to RBC membranes was rapid and readily reversible in a dynamic equilibrium with its binding sites. It is anticipated that this method will be applicable to studies of a wide variety of diseases specifically related to zinc metabolism in humans as well as in animals. (C) 1994 Wiley-Liss, Inc.

Speaking to each other at last? the ghost of tv past, present and to come...

John Hartley discusses TV past, present and future and concludes that 'This brave new world does have a couple of dystopian elements. One is that no-one knows how to fund non-universal TV production. Another is that any future 'imagined community' will have to get used to the fact that most people aren't inside it.

Melbourne on film dossier: 'Ghost Rider'

Despite being set in an unnamed Texan city, Ghost Rider (Mark Steven Johnson, 2007) was a landmark film for Melbourne. It was the first international production to be made at the Central City (now Docklands) Studios, much to the relief of the heavily-invested state government. And it demonstrated to the world (or more importantly, to producers resident in a small part of southern California), that the city was willing and available to be made over and made up to fit even (especially!) the most stupid filmmaking fantasies. Announcing the imminent arrival of the production in October 2004, the Herald Sun boldly predicted that '[c]ity lanes, Telstra Dome and the Yarra River will be the stars of the new action film'. In fact Melbourne is, as the filmmakers intended, (virtually) unidentifiable onscreen. For the Victorian State government, Ausfilm, Film Victoria and the Melbourne Film Office, this lack of specificity was something to be celebrated; a few months before Ghost Rider went in to production, the then State Minister for Innovation, John Brumby, declared that 'it's almost inconceivable that [Ghost Rider] won't put Melbourne on the map internationally'.

Ghost Wash

‘Ghost Wash’ unveils the past in a contemporary context. It is a blending of video projection, sound, music and performance that reconstructs the anger, the angularity, and the angst of Brisbane music from the late 70s through the 80s. The music is contained within an ongoing story about Brisbane music history.

Ghost hunting with lollies, chess and Lego : appreciating the ‘messy’ complexity (and costs) of doing difficult research in education

This paper contributes to conversations about the funding and quality of education research. The paper proceeds in two parts. Part I sets the context by presenting an historical analysis of funding allocations made to Education research through the ARC’s Discovery projects scheme between the years 2002 and 2014, and compares these trends to allocations made to another field within the Social, Behavioural and Economic Sciences assessment panel: Psychology and Cognitive Science. Part II highlights the consequences of underfunding education research by presenting evidence from an Australian Research Council Discovery project that is tracking the experiences of disaffected students who are referred to behaviour schools. The re-scoping decisions that became necessary and the incidental costs that accrue from complications that occur in the field are illustrated and discussed through vignettes of research with “ghosts” who don’t like school but who do like lollies, chess and Lego.

Broken gates and leaky graves : spectral language and Australia's ghost stories

Ghost stories are unusual amongst supernatural literatures in their modelling of a recognisable, mimetic reality interrupted or infiltrated by immaterial forces. In its discussion of Australian ghost stories, this thesis advances a new approach to ghost narratives which seeks to model and articulate the mechanics of ghosts and hauntings as something reliant on and engaged with the material and the mundane.

The rise and fall of the international law of maritime terrorism : the ghost of piracy is still hunting!

Maritime terrorism is a serious threat to global security. A major debate in this regard is the treating of acts of maritime terrorism as piracy by some scholars and a rejection of this view by others. Moreover, the international law of maritime terrorism suffers from fundamental definitional issues, much like the international law of terrorism. This article examines the current international law of maritime terrorism with a particular emphasis on the debate regarding the applicability of the international law of piracy in the case of maritime terrorism. It argues that the international law of piracy is not applicable in the enforcement and prosecution of maritime terrorists on the high seas. International treaties on terrorism and the post-September 11 developments relating to international laws on terrorism have created a workable international legal framework for combating maritime terrorism, despite some bottlenecks.

Interaction of erythrocyte eicosapentaenoic acid and physical activity predicts reduced risk of mild cognitive impairment

Objectives To evaluate relationships between self-reported physical activity, proportions of long-chain omega-3 polyunsaturated fatty acids (LCn3) in erythrocyte content (percentage of total fatty acids) and risk of mild cognitive impairment (MCI) in older adults. Method A cross-sectional study was conducted. Community-dwelling male and female (n = 84) participants over the age of 65 years with and without MCI were tested for erythrocyte proportions of the LCn3s eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). Physical activity was measured using a validated questionnaire. Results The interaction between erythrocyte EPA, but not DHA, and increased physical activity was associated with increased odds of a non-MCI classification. Conclusion An interaction between physical activity and erythrocyte EPA content (percentage of fatty acids) significantly predicted MCI status in older adults. Randomised control trials are needed to examine the potential for supplementation with EPA in combination with increased physical activity to mitigate the risk of MCI in ageing adults.

The ghost in the machine: Legal challenges of neural interface devices

Neural interface devices and the melding of mind and machine, challenge the law in determining where civil liability for injury, damage or loss should lie. The ability of the human mind to instruct and control these devices means that in a negligence action against a person with a neural interface device, determining the standard of care owed by him or her will be of paramount importance. This article considers some of the factors that may influence the court’s determination of the appropriate standard of care to be applied in this situation, leading to the conclusion that a new standard of care might evolve.

Ghost fishing in the pot fishery for blue swimmer crabs Portunus pelagicus in Queensland, Australia

Blue swimmer crabs (Portunus pelagicus) are an economically important crab caught in baited traps throughout the Indo-west Pacific and Mediterranean. In Australia they are traditionally caught using rigid wire traps (approximate to pots) but there has been a recent increase in the use of collapsible pots constructed from polyethylene trawl mesh. Two experiments were conducted in Moreton Bay, Queensland, to determine the ghost fishing potential of lost crab pots on both target and bycatch species and to evaluate the differences between traditional and contemporary pot designs. A lost contemporary, collapsible trawl mesh pot will catch between 3 and 223 R pelagicus per year after the bait has been exhausted, while a traditional wire mesh pot would catch 11-74 crabs peryear. As most fishers now use the collapsible trawl mesh pots, ghost fishing mortality could be as high as 111,811-670,866 crabs per year. Bycatch retention was also higher in contemporary designs. Periods of strong winds appeared to increase the ghost fishing potential of lost pots. The use of escape gaps, larger mesh sizes and construction options that allow for the deterioration of entrance funnels to minimise ghost fishing are recommended to reduce environmental impacts.

Sequence variants in three loci influence monocyte counts and erythrocyte volume

Blood cells participate in vital physiological processes, and their numbers are tightly regulated so that homeostasis is maintained. Disruption of key regulatory mechanisms underlies many blood-related Mendelian diseases but also contributes to more common disorders, including atherosclerosis. We searched for quantitative trait loci (QTL) for hematology traits through a whole-genome association study, because these could provide new insights into both hemopoeitic and disease mechanisms. We tested 1.8 million variants for association with 13 hematology traits measured in 6015 individuals from the Australian and Dutch populations. These traits included hemoglobin composition, platelet counts, and red blood cell and white blood cell indices. We identified three regions of strong association that, to our knowledge, have not been previously reported in the literature. The first was located in an intergenic region of chromosome 9q31 near LPAR1, explaining 1.5% of the variation in monocyte counts (best SNP rs7023923, p=8.9x10(-14)). The second locus was located on chromosome 6p21 and associated with mean cell erythrocyte volume (rs12661667, p=1.2x10(-9), 0.7% variance explained) in a region that spanned five genes, including CCND3, a member of the D-cyclin gene family that is involved in hematopoietic stem cell expansion. The third region was also associated with erythrocyte volume and was located in an intergenic region on chromosome 6q24 (rs592423, p=5.3x10(-9), 0.6% variance explained). All three loci replicated in an independent panel of 1543 individuals (p values=0.001, 9.9x10(-5), and 7x10(-5), respectively). The identification of these QTL provides new opportunities for furthering our understanding of the mechanisms regulating hemopoietic cell fate.

Common variants in TMPRSS6 are associated with iron status and erythrocyte volume

We report a genome-wide association study to iron status. We identify an association of SNPs in TPMRSS6 to serum iron (rs855791, combined P = 1.5 x 10(-20)), transferrin saturation (combined P = 2.2 x 10(-23)) and erythrocyte mean cell volume (MCV, combined P = 1.1 x 10(-10)). We also find suggestive evidence of association with blood hemoglobin levels (combined P = 5.3 x 10(-7)). These findings demonstrate the involvement of TMPRSS6 in control of iron homeostasis and in normal erythropoiesis.