Table_1_In vitro antibacterial activity of antiretroviral drugs on key commensal bacteria from the human microbiota.docx

IntroductionAntiretroviral therapy has improved life expectancy in HIV-infected patients. However, people living with HIV under antiretroviral therapy are at higher risks of developing chronic complications and acquiring multidrug resistant bacteria than healthy population. These factors have been associated with shifts in gut microbiome composition and immune activation. It is unclear how antiretroviral drugs affect gut microbiota composition, but it has been observed that antiretroviral treatment is not able to fully restore gut health after HIV infection. Additionally, some antiretroviral drugs have shown antibacterial activity suggesting that these drugs could have a direct impact on the human microbiome composition. MethodsWe determined the in vitro antibacterial activity of 16 antiretroviral drugs against a set of key clinically relevant and human commensal bacterial strains. ResultsOur results demonstrate that 5 antiretroviral drugs have in vitro antibacterial activity against gut and vaginal human commensal bacteria. Zidovudine has antibacterial activity against Escherichia coli, Klebsiella pneumoniae and Prevotella bivia, abacavir against Gardnerella vaginalis, efavirenz against G. vaginalis and P. bivia and bictegravir against Enterococcus spp. and G. vaginalis. Moreover, we describe for the first time that elvitegravir has antibacterial activity against G. vaginalis and P. bivia and, most importantly, against vancomycin-resistant Enterococcus spp. and methicillin-resistant Staphylococcus aureus strains with MIC values of 4-16 and 4 µg/mL, respectively showing high level of effectiveness against the tested multidrug-resistant bacteria.DiscussionOur results underscore that some antiretroviral drugs may influence the human microbiota composition. In addition, we report the potential use of elvitegravir to treat multidrug-resistant Gram-positive bacteria warranting the need of clinical studies to repurpose this antiretroviral drug. </p

Proposed model of hepcidin in malaria infection.

<p>The regulation of hepcidin in response to infection may vary with cell type. A major response to infection occurs in hepatocytes in response to IL-6. However, our observations support the role of IL-10 in primary macrophages. Availability of iron to erythroid developing cells ultimately depends on macrophages and thus the high concentration of IL-10 may play a key regulatory role. Indeed, actively dividing cells like those found in the bone marrow are more susceptible to oxidative damage. In this context, both the direct anti-inflammatory effect of IL-10 and its indirect effect on iron restriction through the up-regulation of hepcidin may be beneficial.</p

Effect of IL-10 and IL-6 on the regulation of hepcidin in (a) primary macrophages and (b) HepG2 cells. Data show fold-increase relative to mRNA in media control.

<p>Error bars indicate standard error of the mean. Bar plots show data from at least 3 independent experiments. ** <i>P</i><0.01, ***<i>P</i><0.001 (Mann-Whitney test, compared with Media control).</p

Clinical features in Gambian children with severe malaria independently associated with a fatal outcome.

<p>The multiple logistic regression analysis included 1,931 observations with complete data (5 degrees of freedom) χ² = 180.4 (P<0.001); pseudo-R2 = 0.10; Goodness-of- fit, statistics: Hosmer-Lemeshow  = 4.47 (P = 0.61). AUC =  area under the curve.</p

Baseline Characteristics of the Study Population.

<p>Baseline Characteristics of the Study Population.</p

Prevalence of clinical features in Gambian children admitted to hospital with severe malaria.

<p>variables were defined as follows: Prostration, inability to sit (children aged >7 months); Impaired consciousness, BCS ≤4; Coma, BCS ≤2; Repeated convulsions, >3 in 24 h; Severe anemia (with any parasite density), Hb <5 g/dL or PCV <15; Respiratory distress, abnormal respiratory pattern (respiratory pattern values > or  = 3), grunting or use of accessory muscles of respiration, or abnormally deep (acidotic) breathing; Hypoglycemia ≤2.2 mM; Hyperlactatemia, plasma lactate >5 mM; Hyperpyrexia, temp>40°C; Hyperparasitemia, <i>P. falciparum</i> parasite density >500,000 /µl; Hypotensive shock, circulatory collapse with systolic blood pressure <50 mmHg; Hepatomegaly >2 cm below right costal margin; Splenomegaly >2 cm below left costal margin.</p

Clinical features associated with death in children with SM.

<p>Odds of death and blood lactate concentration in children with SM. Data show the odds ratio (95%CI) of death in relation to increasing concentrations of blood lactate in 467 children with SM (a). Odds of death and Blantyre coma score. OR and P values are relative to BCS = 5 (b). Specificity and sensitivity of different blood lactate concentration cut-off values (c) and coma scores measured by BCS (d) to predict death. AUC =  area under the curve,* P<0.05, ***P<0.001.</p

Baseline clinical characteristics of the study population.

<p>Values are presented as median [interquartile range]. ADMA: asymmetric dimethylarginine, sVCAM: soluble vascular cell adhesion molecule, PfHRP2: <i>P</i>. <i>falciparum</i> histidine-rich protein 2.</p><p>^a p < 0.001 compared to healthy Gambian children by Mann-Whitney test.</p><p>^b p < 0.001 compared to uncomplicated malaria by Mann-Whitney test.</p><p>Baseline clinical characteristics of the study population.</p

Multiple linear regression analysis of the relationships between ADMA and arginine and hemoglobin, HRP2, sVCAM, or lactate.

<p>ADMA, Arg, HRP2, and sVCAM were natural log-transformed. Lactate was square root-transformed. ADMA and arginine were explanatory variables in four separate linear models predicting hemoglobin, HRP2, sVCAM, or Lactate.</p><p>Multiple linear regression analysis of the relationships between ADMA and arginine and hemoglobin, HRP2, sVCAM, or lactate.</p

The ADMA/arginine ratio is acutely elevated in African children with severe malaria.

<p>ADMA and arginine concentrations were measured in plasma samples collected at the time of presentation (Day 0) and at follow-up visits 28 days later (Day 28) in children with WHO-defined uncomplicated malaria or severe malaria. Healthy Gambian children served as a reference group. Wilcoxon test was used for pair-wise comparison of admission and day 28 mesurements within individuals (47 paired observations from patients with severe malaria; 65 paired observations from patients with uncomplicated malaria). Mann-Whitney test was used to compare patients with severe malaria (n = 81) versus uncomplicated malaria (n = 75) and to compare patients with uncomplicated malaria versus healthy children (n = 31). Each horizontal line depicts the group median. **** p < 0.0001; ns p > 0.05.</p