Characteristics of the study population according to institutionalization status.

<p>ADL = Activities of Daily Living <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0046061#pone.0046061-Katz1" target="_blank">[30]</a>; DSSI = Duke Social Support Index <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0046061#pone.0046061-Koenig1" target="_blank">[10]</a>; IADL = Instrumental Activities of Daily Living <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0046061#pone.0046061-Fillenbaum1" target="_blank">[31]</a>; PASE = Physical Activity Scale for the Elderly <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0046061#pone.0046061-Washburn1" target="_blank">[11]</a>.</p>a<p>Missing data not included in percentages.</p

The percentage of participants institutionalized with increasing age.

<p>Test for deviation from linear trend: P = 0.0003.</p

Final multivariate models for predictors of institutionalization up to 3.4 years and beyond 3.4 years of follow-up.

<p>ADL = Activities of Daily Living <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0046061#pone.0046061-Katz1" target="_blank">[30]</a>; ESB = English Speaking Background; IADL = Instrumental Activities of Daily Living <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0046061#pone.0046061-Fillenbaum1" target="_blank">[31]</a>;</p><p>MCI = Mild Cognitive Impairment; NESB = Non-English Speaking Background.</p><p>NA, not applicable = not a significant predictor during the time period.</p

Kaplan-Meier survival curves of the time until institutionalization by cognitive status groups.

<p>Kaplan-Meier survival curves of the time until institutionalization by cognitive status groups.</p

The Effects of Dietary Macronutrient Balance on Skin Structure in Aging Male and Female Mice

<div><p>Nutrition influences skin structure; however, a systematic investigation into how energy and macronutrients (protein, carbohydrate and fat) affects the skin has yet to be conducted. We evaluated the associations between macronutrients, energy intake and skin structure in mice fed 25 experimental diets and a control diet for 15 months using the Geometric Framework, a novel method of nutritional analysis. Skin structure was associated with the ratio of dietary macronutrients eaten, not energy intake, and the nature of the effect differed between the sexes. In males, skin structure was primarily associated with protein intake, whereas in females carbohydrate intake was the primary correlate. In both sexes, the dermis and subcutaneous fat thicknesses were inversely proportional. Subcutaneous fat thickness varied positively with fat intake, due to enlarged adipocytes rather than increased adipocyte number. We therefore demonstrated clear interactions between skin structure and macronutrient intakes, with the associations being sex-specific and dependent on dietary macronutrient balance.</p></div

Macronutrient intake influences subcutaneous fat.

<p>Response surfaces showing the association of macronutrient intake (protein, carbohydrate and fat in kJ/d) on subcutaneous adipocyte size (μm²) and adipocyte numbers (cells/10⁵μm²). (a-c) male adipocytes become grossly enlarged with high fat intake whist adipocytes proliferate with high protein intake (d-f; cells/10⁵μm²). (g-i) female adipocytes enlarge to a lesser extent than male adipocytes with high carbohydrate or fat intake and proliferate with increasing protein intake (j-i). For each 2D slice, the third factor is at its median. The red line indicates the ratio of macronutrients that minimizes each response. (see also <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0166175#pone.0166175.s005" target="_blank">S4 Table</a>)</p

The association between protein intake and male and female skin structure.

<p>H&E staining for male mouse skin layers (a-c) and female mouse skin layers (d-f), x20 magnification, scale bar = 200 μm, ‘d’ indicates area of dermis and ‘s’ indicates area of subcutaneous fat. High protein intake significantly increases male dermis thickness and thins the subcutaneous fat. In females, no effect of protein intake on skin structure was identified. Dietary composition of standard chow is protein (21%), carbohydrate (63%) and fat (16%). Mean skin thickness (a) d = 391 μm, s = 54 μm, (b) d = 275 μm s = 90 μm, (c) d = 228 μm, s = 171 μm, (d) d = 203 μm, s = 148 μm, (e) d = 127 μm, s = 233 μm, (f) d = 194 μm, s = 173 μm.</p

Dermis thickness (μm) and subcutaneous fat thickness (μm) are inversely proportional and correlate with body fat%.

<p>Dermis thickness increases with a thinner subcutaneous fat in both (a) males (R² = -0.448; P<0.001) then (b) females (R² = -0.362; P<0.001). Subcutaneous fat increases with increasing body fat % in (c) male and (d) female mice (R² = 0.549; P<0.001 and R² = 0.626; P<0.001, respectively).</p

Effects of manipulating rafts on porosity.

<p>The effects of 7KC, Triton X-100 and cytochalasin D on the porosity and diameter of fenestrations (determined using SEM) and generalized polarization (GP, following staining with LAURDAN imaged using two-photon microscopy) (* significantly different from control values P<0.05, each data point represents average ± SEM of 7–28 images and 83–2840 fenestrations).</p

Visualization of membrane rafts and fenestrations.

<p>(A–C) 3D-SIM of LSECs stained with Bodipy FL C5 ganglioside GM1, a marker for rafts (green) and Cell-Mask Orange, a cell membrane marker (orange). There is an inverse distribution between liver sieve plates and membrane rafts. Membrane rafts are mostly perinuclear while sieve plates are mostly peripheral. Some sieve plates are identified by an asterix (*) and fenestrations can be resolved within the sieve plates. Rafts are shown with arrows (→). The areas marked in a box (clustered rafts) are further magnified in (D–G). (D–G) Magnification of areas in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0046134#pone-0046134-g001" target="_blank">Figure 1(A–B)</a> showing clustered membrane rafts with raised perimeters. (H) TIRFM of LSEC stained with NBD-cholesterol (green), a marker for rafts, and CellMask Orange (orange) showing perinuclear distribution of rafts (arrows). Fenestrations are not resolved within the sieve plates (*) with TIRFM. (I) TIRFM of LSEC stained with Bodipy FL C5 ganglioside GM1, a marker for rafts, and CellMask Orange (orange) confirming mostly perinuclear distribution of rafts. Scale bar 5 µm (A, B, H, I), 1 µm (C–G).</p