Box Plots of Cysteine Levels in Bronchoalveolar Lavage between non-HIV and HIV-infected Subjects.

<p>Box Plots of Cysteine Levels in Bronchoalveolar Lavage between non-HIV and HIV-infected Subjects.</p

Multiple Linear Regression Model for BAL Cysteine.

<p>P = 0.76.</p><p>Adjusted R² = −0.05.</p

Demographic characteristics of HIV-infected Subjects Enrolled in the Study.

<p>BMI =  Body Mass Index.</p><p>SMAST =  Short Michigan Alcohol Screening Test.</p><p>AUDIT =  Alcohol Use Disorders Identification Test.</p><p>ART =  Anti-retroviral medications.</p

Box Plots of Glutathione Levels in Bronchoalveolar Lavage HIV-infected Subjects on and off Anti-retroviral Therapy.

<p>Box Plots of Glutathione Levels in Bronchoalveolar Lavage HIV-infected Subjects on and off Anti-retroviral Therapy.</p

Box Plots of Glutathione Levels in Bronchoalveolar Lavage between non-HIV and HIV-infected Subjects.

<p>Box Plots of Glutathione Levels in Bronchoalveolar Lavage between non-HIV and HIV-infected Subjects.</p

Chronic ethanol ingestion modifies Cys thiols on α-ENaC.

<p>A) <u>Left panel</u> F5M labeling of α-ENaC protein immuno-precipitated from chronic ethanol fed mice (left lane) or maltodextrin control animals (right lane). <u>Middle panel</u> shows bioluminescent detection of α-ENaC using the same membrane shown in left panel. <u>Right panel</u> Co-registration of F5M and ENaC signal (ENaC signal is pseudo-colored and F5M signal contrast reduced in order to enhance co-registration of data obtained from the same blot). B) Normalization of F5M RLU to ENaC expression levels; n = 3; p<0.05.</p

Acute EtOH consumption increases ROS production in C57/Bl6 mice.

<p>Top panels (A–C) show excised lung from control animals, or mice ingesting 20% EtOH solution for 3 days, following instillation of 200 µL DHE solution. A = 510/620 nm excitation emission (red signal); 30 sec exposure. B = panel A co-registered with x-ray image C = Additive primary color (RGB) image of lungs excised en bloc (showing trachea, heart, and lung. Panels D (510/620 nm; 10 min exposure) and E (510/620 nm co-registered with X-ray) show excised lung from control and EtOH mice following instillation of 200 µL vehicle control. Intensity bars, and rhodamine positive signal controls included for B–E. F) Quantification of homogenized lung samples (shown in A–E) using a microplate reader assay (n = 12 from 3 animals; p<0.05).</p

Multichannel confocal imaging shows EtOH mediated changes in Rac1 expression and subcellular localization in the lung.

<p>Anti-Rac1 antibody detected using Alexa 488 conjugated secondary antibody (488/519 nm; green fluorescence); nuclei were stained DAPI (350/470 nm; blue fluorescence); and plasma membrane labeled with Cell Mask Deep Red (649/666 nm). <u>Right panel</u>: white light image of lung slice preparation and Rac1 labeling; <u>middle panel</u>: Rac1 localization relative to DAPI stained nuclei; <u>left panel</u>: Rac1 co-localization with Deep Red labeled plasma membrane. Pixels containing both red and green color contributions produce various shades of orange and yellow indicate Rac1 co-localization with the plasma membrane. Subsets represent z-axis obtained from horizontal and vertical regions as indicated.</p

Acute 0.16% ETOH treatment increases sodium channel activity.

<p>A) Continuous single channel recording obtained from primary isolated rat T2 cell treated with 0.16% EtOH after 5 min control recording period as indicated with enlarged portions of the trace. Arrow indicates closed state of channels, with downward deflections from arrow indicating Na movement into the cell. B) Point amplitude histograms show frequency of NSC and HSC activity in representative recording before and after EtOH treatment. C) Conductances (γ) of representative HSC (4.9pS) and NSC (11pS) channels shown in representative trace. D) Number and open probability (NPo) of ENaC reported before and after 0.16% EtOH treatment in 7 cell-attached recording; p<0.05. E-F) I/V curve of all cell recordings with average HSC γ = 6.1 and NSC γ = 22.2.</p

LPS inoculation and chronic EtOH ingestion enhances the rate of alveolar fluid clearance.

<p>A) In vivo assessment of lung fluid volumes after saline challenge in EtOH (n = 13), isocaloric control (maltodextrin, n = 13) and C57Bl/6 mice (n = 10) indicate similar patterns of lung fluid clearance; albeit an observed delay in clearance in EtOH mice compared to maltodextrin between 3.5–4 hrs following saline challenge. B) In vivo assessment of 1 mg/mL LPS inoculated animals chronically fed an EtOH diet (n = 4) cleared fluid at faster rates compared to isocaloric control groups of animals fed maltodextrin (n = 5) between 190–220 min following saline challenge (as indicated by solid line, p<0.05). LPS inoculation of EtOH animals showed significantly elevated rates of alveolar fluid clearance compared to LPS inoculated C57Bl/6 mice beginning at 20 minutes following saline challenge (denoted by dashed line, p<0.05). C) Co-instillation of 1 µM NSC23766 and LPS in chronic ethanol mice indicates that small G protein Rac1 plays an important role in alcohol lung and alveolar fluid balance. Animals instilled with LPS and NSC23766 (n = 5) failed to clear saline challenge to the extent observed in LPS (n = 4) inoculated EtOH animals; p<0.05 as indicated.</p