Demographics and Cognitive Summary by ε2.

<p>All means are reported ± 1 standard error. All <b>bolded</b> parameters were significant according to ANOVA, Chi-square analysis, or Fisher’s Exact Test as appropriate. CN = cognitively normal, MCI = mild cognitive Impairment, AD = Alzheimer’s Disease, ε2 = APOE epsilon 2.</p

Freesurfer hippocampal region of interest.

<p>Coronal, sagittal, and axial (1.5T) T1-weighted MR images from the Freesurfer Image Analysis Suite of sample subject “Bert”, with color overlays depicting the extent of the Freesurfer hippocampal region of interest used to derive hippocampal volumes.</p

<i>Leishmania donovani</i> Infection Enhances Lateral Mobility of Macrophage Membrane Protein Which Is Reversed by Liposomal Cholesterol

<div><p>Background</p><p>The protozoan parasite <i>Leishmania donovani</i> (LD) reduces cellular cholesterol of the host possibly for its own benefit. Cholesterol is mostly present in the specialized compartment of the plasma membrane. The relation between mobility of membrane proteins and cholesterol depletion from membrane continues to be an important issue. The notion that leishmania infection alters the mobility of membrane proteins stems from our previous study where we showed that the distance between subunits of IFNγ receptor (R1 and R2) on the cell surface of LD infected cell is increased, but is restored to normal by liposomal cholesterol treatment.</p><p>Methodology/Principal Findings</p><p>We determined the lateral mobility of a membrane protein in normal, LD infected and liposome treated LD infected cells using GFP-tagged PLCδ1 as a probe. The mobility of PLCδ1 was computationally analyzed from the time lapse experiment using boundary distance plot and radial profile movement. Our results showed that the lateral mobility of the membrane protein, which is increased in infection, is restored to normal upon liposomal cholesterol treatment. The results of FRAP experiment lent further credence to the above notion. The membrane proteins are intimately linked with cellular actin and alteration of cellular actin may influence lateral mobility. We found that F-actin is decreased in infection but is restored to normal upon liposomal cholesterol treatment as evident from phalloidin staining and also from biochemical analysis by immunoblotting.</p><p>Conclusions/Significances</p><p>To our knowledge this is the first direct demonstration that LD parasites during their intracellular life cycle increases lateral mobility of membrane proteins and decreases F-actin level in infected macrophages. Such defects may contribute to ineffective intracellular signaling and other cellular functions.</p></div

Confocal images of <i>plcδ1-gfp</i> transfected RAW 264.7 cells followed by infection with LD (I-MΦ).

<p>The expression of PLCδ1-GFP (green) is on the cell surface. The cells were stained with Hoechst 33342 (blue). Parasites are marked by blue dot and indicated by arrow.</p

Demographic and cognitive summary by ε4.

<p>All means are reported ± 1 standard error. All <b>bolded</b> parameters were significant according to ANOVA, Chi-square analysis, or Fisher’s Exact Test as appropriate. CN = cognitively normal, MCI = mild cognitive Impairment, AD = Alzheimer’s Disease, ε4 = APOE epsilon 4.</p

Summary of effects for variables on hippocampal volume: <i>APOE</i> dose-dependent model.

<p>n = 662.</p><p>Results for model which treats APOE epsilon “X” allele effects as dose-dependent effects. APOE effect sizes of each variable are reported in terms of mm³/(unit variable), e.g., the effect of “Age” on the outcome variable, hippocampal volume, is −23.31 mm³/year of age. “:” denotes interaction terms between variables. Reference for the model is an APOE ε3/ε3 CN individual. Note the lack of a main effect of APOE epsilon 4 (“ApoE4”) in the model and thus in CN subjects.</p

Summary of unadjusted hippocampal volumes by <i>APOE</i> ε4 allele dose and diagnostic category.

<p>Unadjusted values. “ε4 dose” = copy number of APOE ε4 allele,</p><p>“SD” = standard deviation, “SE” = standard error of the mean. “Mean” units are mm³.</p

Radial distance map of a cell.

<p>A) Blue line shows cell boundary (radial distance = 1). Red line show contours of equal radial distance within the cell (radial distance <1). Green lines show contours of equal radial distance outside the cell (radial distance >1). B) Plot of the distribution of PLCδ1-GFP expression as a function of distance from center of the cell. The red line shows the average activity as a function of distance, obtained via a smoothing line. The x-axis (radial distance) is truncated at 1.5 to avoid contamination of PLCδ1-GFP expression from other cells.</p

Analysis of PLCδ1 movement.

<p>RAW264.7 cells were transfected with <i>plcδ1-gfp</i> plasmid. Average motions of the PLCδ1-GFP were measured by computational analysis as a function of distance from center of cell. Each line represents a particular cell. Motion is on a scale of 0–6 (arbitary unit, a.u.). A) N-MΦ; B) I-MΦ; and C) I-MΦ-CL.</p

Summary of unadjusted hippocampal volumes by <i>APOE</i> ε2 allele dose and diagnostic category.

<p>Unadjusted values. “ε2 dose” = copy number of APOE ε2 allele,</p><p>“SD” = standard deviation, “SE” = standard error of the mean. “Mean” units are mm³.</p