1,611 research outputs found

    The effect of changes in plasma membrane lipid composition on the heat sensitivity of Hepatoma tissue culture cells and selected plasma membrane enzymes

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    Hepatoma Tissue Culture (HTC) cells grown in the presence of 60µM arachidonic acid for 24, 36 and 48 hours became progressively more thermosensitive than control cells. However, this difference in thermal sensitivity was only detectable with the clonogenic assay and not with the colorimetric assay. Attempts were also made to manipulate cellular cholesterol levels. Firstly, some cells were incubated with phosphatidylcholine liposomes to deplete the plasma membrane of cholesterol: Secondly, another group of cells were treated with 25 hydroxycholesterol, an inhibitor of cholesterol synthesis, to lower cholesterol levels: Finally, a third group of cells were supplemented with cholesterol hemisuccinate, a hydrophilic ester of cholesterol. The first two approaches did not enhance the thermal sensitivity of HTC cells. Supplementation with cholesterol hemisuccinate, which was predicted to partition in to the plasma membrane and reduce membrane fluidity, resulted in increased thermal sensitivity of the cells. Thus, the thermal sensitivity of HTC cells could be enhanced by supplementation with either arachidonate or cholesterol hemisuccinate. A rapid plasma membrane isolation procedure was developed which generated plasma membranes in relatively high yield and purity. The plasma membrane- enriched fraction was also assayed for contaminating intracellular membranes by determining marker enzyme activities associated with these membranes. Using this method, plasma membranes were prepared from HTC cells grown m 60µM arachidonic acid for 36 hours and from cells grown in normal medium. Analysis of the plasma membrane showed that the arachidonic acid content of the phospholipid fatty acyl groups had been significantly increased in cells grown in the presence of this fatty acid. There was no change in the cholesterol/phospholipid molar ratio or cholesterol concentration relative to amount of protein in the plasma membranes from the two cell populations. The measurement of fluidity using DPH fluorescence polarisation revealed that the increase in the arachidomc acid content of the plasma membrane phospholipid acyl groups was associated with enhanced plasma membrane fluidity when compared to control plasma membranes. This increase in plasma membrane fluidity correlated with the enhanced thermal sensitivity of the cells grown in arachidonic acid-containing medium when compared to cells grown in normal medium. Furthermore, the thermal sensitivity of Na(^+), K(^+) –ATPase and alkaline phosphodiesterase I were assessed in plasma membranes derived from arachidonic acid-supplemented and control cells. The enhanced fluidity of plasma membranes derived from arachidonate-supplemented cells also correlated with increased thermosensitivity of alkaline phosphodiesterase I

    Second Generation Immigrant Adaptation: Construction of a Hybrid Cultural Identity

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    This study uses a postcolonial perspective to examine the construction of cultural identities in second generation South Asian women. It critiques traditional strategies of immigrant incorporation, including assimilation and cultural pluralism, for their androcentric and essentialist tendencies. It was found that the women constructed a cultural hybrid identity, and using Homi Bhabha\u27s notion of third space, I discuss the process of how this hybrid identity is constructed. A phenomenological approach, in which the subjective voices of the participants are privileged, was used to analyze nine interviews for themes relating to the construction of a hybrid identity

    Evaluation of an open source method for calculating physical activity in dogs from harness and collar based sensors

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    Abstract Background The ability to make objective measurements of physical activity in dogs has both clinical and research applications. Accelerometers offer a non-intrusive and convenient solution. Of the commercialy available sensors, measurements are commonly given in manufacturer bespoke units and calculated with closed source approaches. Furthermore, the validation studies that exist for such devices are mounting location dependant, not transferable between brands or not suitable for handling modern raw accelerometry type data. Methods This paper describes a validation study of n = 5 where 4 sensors were placed on each dog; 2 on a harness and 2 on a collar. Each position held two sensors from different manufacturers; Actigraph (which has previously been validated for use on the collar) and VetSens (which provides un-filtered accelerometry data). The aims of the study was to firstly evaluate the performance of an open-design method of converting raw accelerometry data into units that have previously been validated. Secondly, comparison was made between sensors mounted at each location for determining physical activity state. Results Once the raw actigraphy data had been processed with the open-design method, results from a 7 day measurement revealed no significant difference in physical activity estimates via a cutpoint approach between the sensor manufacturers. A second finding was a low inter-site variability between the ventral collar and dorsal harness locations (Pearsons r2 = 0.96). Conclusions Using the open-design methodology, raw, un-filtered data from the VetSens sensors can be compared or pooled with data gathered from Actigraph sensors. The results also provide strong evidence that ventral collar and dorsal harness sites may be used interchangeably. This enables studies to be designed with a larger inclusion criteria (encompassing dogs that are not well suited for wearing an instrumented collar) and ensures high levels of welfare while maintaining measurement validity
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