Graph showing the percent recoveries of viable microorganisms following our sample preparation protocol.

<p>Data are averages of several replicates (n>3).</p

Results from PCR.

<p>A: PCR data show that pellets from whole blood samples (10 mL) spiked with higher concentrations of MSSA amplify sooner (pellet volume in PCR well = 8 μL). B: Contingency table for PCR analysis of processed positive (100 CFU/mL) and negative (0 CFU/mL) whole blood samples.</p

List of microorganisms used in the current study and their cultivation conditions.

<p>^<i>a</i> Gram-positive bacterium;</p><p>^<i>b</i> Gram-negative bacterium;</p><p>^<i>c</i> Yeast</p><p>List of microorganisms used in the current study and their cultivation conditions.</p

Images of the custom-made microbial concentration devices.

<p>Left: a blown-up view of the individual parts of the device. Right: a CAD model of a fully assembled device.</p

Images of the pellets after lysing 10 mL of whole blood.

<p>A: Microscopic view of an unspiked blood sample after lysis process at 60x magnification; B: For reference, microscopic view of unlysed whole blood at 60x magnification; C: Front view of the pellets; D: Side view of the pellets; E: For reference, from left to right, 2.5, 5.0, 10, and 15 μL of whole blood.</p

Viral RNA and neuronal markers (NAA/Cr, MAP2, and SYN) with and w/o treatments.

<p>A. The decrease in NAA/Cr was arrested with this regimen of combination antiretroviral treatment as well as minocycline treatment resulting in higher NAA/Cr levels when compared to untreated animals sacrificed at 8 wpi, (cART P = 0.09; MN_persistent P = 0.029; MN_short P = 0.0012). Animals that were minocycline treated and had partial immune reconstitution of the CD8 T cell population showed the best results. B. MAP2 levels in combination antiretroviral treated and in minocycline treated animals were significantly higher than untreated SIV-infected animals (cART P = 0.006; MN_persistent P = 0.010; MN_short P = 0.016). C. Similarly, SYN levels in combination antiretroviral treated animals were significantly higher than untreated SIV-infected animals (P = 0.002). (* indicates statistically significant differences when compared to untreated animal.).</p

Correlation between viral loads in different compartments and CD14+/CD16+ monocytes.

<p>Correlation between viral loads in different compartments and CD14+/CD16+ monocytes.</p

MRS and IHC show neuronal injury in infected animals w/o treatment.

<p>A. The neuronal marker N-Acetylasparate/creatine steadily declined following SIV infection in all animals, reaching decreased as low as 20% below baseline by 8 wpi in untreated animals. (* indicates significant changes from baseline levels prior infection.) B. Average microtubule-associated protein 2 levels in the frontal cortex of SIV+ untreated animals sacrificed at 4 wpi, 6 wpi and 8 wpi were lower compared to uninfected control animals. (* indicates significant differences from controls.) C. Average synaptophysin levels in the frontal cortex of SIV+ untreated animals sacrificed at 4 wpi, 6 wpi and 8 wpi were lower compared to uninfected control animals. (* indicates significant differences from controls).</p

Viral levels in the three compartments and activated monocytes with and w/o treatment.

<p>A. Treated animals revealed lower plasma vRNA levels (~1 log) at endpoints when compared to untreated animals at 8 wpi (cART P = 0.019; MN_persistent P = 0.0063). Lowest plasma SIV RNA levels (~2 log compared to untreated) were observed in animals that were minocycline treated and had partial immune reconstitution of the CD8⁺ T cell population (P = 0.0032). (* indicates statistically significant differences when compared to untreated animals.) B. Cerebrospinal fluid viral levels decreased in cART and in minocycline treated animals after endpoint, yet differences in cerebrospinal fluid viral burden failed to reach significance. C. Viral levels in cART and minocycline treated animals were lower than those in untreated animals (cART P = 0.007; MN_persistent P = 0.006; MN_short P< 0.0001). Short-term CD8-depleted minocycline-treated animals had significantly reduced brain vRNA levels when compared to all other cohorts, specifically in animals that were minocycline treated and CD8-depleted (P = 0.032). (* indicates statistically significant differences when compared to untreated animals. # indicates statistically significant differences when compared to all other cohorts.) D. The cART cohort and the minocycline treated cohort had significantly lower CD14⁺/CD16⁺ monocytes compared to untreated at their endpoints (cART P = 0.014; MN_persistent P = 0.004; MN_short P = 0.0035). (* indicates statistically significant differences when compared to untreated animal.).</p

Viral RNA plateaus w/o treatment.

<p>A. Viral RNA was detectable in plasma 6 days after infection (~10⁷ copies eq./mL) and the viral load approaches a plateau by 4 weeks after infection. The mean plasma viral load was 4.9 x 10⁸ copies eq./mL at 8 wpi. Cerebrospinal fluid viral loads were found to be approximately 3 orders of magnitude lower than plasma. B. The amount of viral RNA in the frontal cortex of untreated animals was 7.5 x 10⁵, 9.2 x 10⁵ and 3.0 x 10⁵ copies eq./g at 4, 6, and 8 weeks after infection, respectively, and was not significant (P > 0.11) and had a mean of ~ 10⁶ copies eq./g. C. SIV+ animals had high levels of circulating CD14⁺/CD16⁺ monocytes 28 days after infection, and they had ~300 cells/μL by 8 weeks. (* indicates significant differences from pre-infection).</p