ddPCR detection of HIV-1 HXB2 and HIV-1 RM virus DNA and Cell-associated RNA from NHBE cell cultures.

<p>ddPCR detection of HIV-1 HXB2 and HIV-1 RM virus DNA and Cell-associated RNA from NHBE cell cultures.</p

Patients with HIV have less E-cadherin in their lung epithelium.

<p><b>A.</b> There is a slight decrease in steady state E-cadherin mRNA as detected by RT-qPCR. (n = 5, *p<0.05, student T-test) <b>B</b>. Pretreating NHBE cells for 1 hour with the lysosomal inhibitors bafilomycin or chloroquine abrogated the reduction in E-Cadherin that was seen following exposure to an X4 tropic virus (IIIB) or dual tropic virus (RF). (n = 4, *p<0.05, Anova with Bonferroni correction) <b>C.</b> Pretreating NHBE cells cells with the lysosomal inhibitors bafilomycin or chloroquine abrogated the increased permeability that was seen following exposure to the X4 tropic virus IIIB (compared to vehicle-only control which was 1:1000 DMSO in media). (n = 4, *p<0.05, Anova with Bonferroni correction).</p

p24 is detected in the basolateral media following exposure to HIV-1.

<p>p24 antigen levels measured from the basolateral media of NHBE cells infected with HXB2 or RM virus at days 4, 6, 10 and 14 post-infection. p24 levels decreased between day 4 and day 14 for both HXB2 (from 0.296±0.002 [Mean±SD] at day 4 to 0.033 ± 0.004 at day 14) and RM (from 0.295±0.003 at day 4 to 0.000 at day 14) infected NHBE cells (n = 3).</p

X4 tropic virus disrupts cell-cell adhesion and increases pro-inflammatory signaling leading to increased barrier permeability.

<p><b>A.</b> The airway epithelium is formed by aggregates of ciliated cells that are joined together by adhesive proteins like E-cadherin. <b>B.</b> Our data indicates that X4 tropic viruses are able to bind to the CXCR4 receptor on the airway epithelial cells and enter these cells. Binding of X4 virus to the CXCR4 receptor leads to increase in pERK. <b>C.</b> Binding X4 tropic virus to the CXCR4 receptor decreases E-cadherin, increases monolayer permeability, and increases the expression of ICAM-1 contributing to tissue remodeling.</p

X4 tropic virus increases abundance of ICAM-1 and pERK.

<p><b>A.</b> Following exposure to X4 tropic virus IIIB there was a statistically significant increase in the abundance if ICAM-1 that was not seen following exposure to R5 tropic virus BAL. (n = 3, *p<0.05, Anova with Bonferroni correction) <b>B.</b> After exposure to HIV-1 there was an immediate increase in the active phosphorylated ERK but no change in total ERK (n = 4, *p<0.05, Anova with Bonferroni correction).</p

X4 tropic virus and dual tropic virus increase airway epithelial permeability.

<p><b>A.</b> NHBE cells were exposed in the basolateral compartment to either an X4 tropic virus or an R5 tropic virus and after a four hour exposure to an X4 tropic virus, IIIB, there was a statistically significant increase in barrier permeability as measured by FITC Dextran that was not seen after four hours of exposure to an R5 tropic virus, BAL. At lower virus titers (0.05ng/ml or 0.5ng/ml), IIIB caused an increase in epithelial permeability although these changes were not statistically significant. (n = 22, *p<0.05, Anova with Bonferroni correction) <b>B.</b> Lower titers of IIIB virus (0.5ng/ml) were able to cause statistically significant increases in epithelial permeability with a 24 hour exposure. (n = 14, *p<0.05, Anova with Bonferroni correction) <b>C.</b> Both X4 tropic virus, MN and HXB2, and dual tropic virus, RF, (all at concentrations of 5ng/ml) caused similar changes in barrier permeability after four hours of exposure (n = 10, *p<0.05, Anova with Bonferroni correction).</p

HIV reduces the abundance of E-Cadherin.

<p><b>A.</b> Immunofluorescence shows a reduction in E-Cadherin along the basolateral membrane (white arrow) after exposure to X4 tropic virus IIIB at a concentration of p24 (5ng/ml) which was not seen after exposure to R5 tropic virus BaL at the same concentration. <b>B.</b> Western blot probing for E-Cadherin at a dilution of 1:1000 shows marked reduction in abundance of E-Cadherin following four hour basolateral exposure to X4 tropic virus but not R5 tropic virus. (n = 6 BaL and 10 IIIB samples, *p<0.05, Mann-Whitney rank sum test) <b>C.</b> By Western analysis, there is a marked reduction in E-cadherin in an epithelial cell whole cell lysate from a patient with HIV and COPD when compared to that obtained from an HIV negative COPD negative individual and an HIV positive COPD negative individual. <b>D.</b> While in the control patients, there is a normal distribution of E-cadherin level, there is a suggestion of two subpopulations of E-cadherin abundance in HIV patients. One with higher E-Cadherin levels and one with lower E-Cadherin levels. While study patients did not have viral tropism studies performed, the dots circled in red represents patients on the CCR5 inhibitor miraviroc suggesting R5 tropic disease. (Data compared using Kruskal-Wallis assessment).</p

Additional file 1: of Short Physical Performance Battery and all-cause mortality: systematic review and meta-analysis

Short Physical Performance Battery and all-cause Mortality: Systematic Review and Meta-analysis. eTable 1. New-Castle Ottawa Scale for quality assessment. eTable 2. Source for follow-up of all the studies included in the meta-analysis. eTable 3. Meta-regression analyses considering population characteristics of each study included in the meta-analysis. eTable 4. Assessment of publication bias. eTable 5. PRISMA checklist. eFigure 1. Funnel plot and Trim and Fill analysis. A. Relation between SPPB 0-3 vs 10-12 and all-cause mortality. B. Relation between SPPB 4-6 vs 10-12 and all-cause mortality. C. Relation between SPPB 7-9 vs 10-12 and all-cause mortality. eFigure 2. Scatter Plot of meta-regression analysis for female sex, diabetes mellitus and age and relation between SPPB 7-9 vs 10-12 and all-cause death. (DOCX 146 kb

Life expectancy estimates (LEE) and standard errors (SE) at age 20 years using unweighted mortality rates, by calendar period, 2000 to 2007.

<p>Note: LEE, <i>life expectancy estimate (years)</i>; SE, <i>standard error</i>; ART, <i>combination antiretroviral therapy</i>.</p

Mid-point life expectancy estimates at age 20 years in three calendar periods, overall and by sociodemographic characteristics, 2000–2007.

<p>Panel A: Life expectancy at age 20 years, overall. Panel B: Life expectancy at age 20 years, by sex. Panel C: Life expectancy at age 20 years, by transmission group. Panel D: Life expectancy at age 20 years, by race. Panel E: Life expectancy at age 20 years, by CD4 cell count (cells/mm³) at ART initiation.</p