Kosta papic biography of william

kosta papic biography of william
Lipid peroxidation d , DNA oxidation e and cell death f were assessed at indicated time points post-infection p. Together these findings support the concepts that TB is associated with excessive oxidative stress and death in infected macrophages and show that this response can be successfully reduced by treatment with NAC.

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kosta papic biography of william

Middlebrook 7H9 alone was used as a negative control. Lipid peroxidation in plasma and culture supernatants was quantified using an assay kit from Cayman Chemical, which measures the formation of malondialdehyde MDA.

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Results from this assay were shown as concentration of 8-OH-DG. Cell death was plotted as percentage of cell death compared with positive control H 2 O 2. The cells used for the ROS measurement assay were previously detached from the culture plates using trypsin 0. Results were plotted as histograms where the mean fluorescence intensity MFI was compared between the experimental groups.

Kosta Papić

Mice were infected with H37Rv M. The dose of NAC used was based on studies reported previously, which describe the effect of NAC treatment during chronic Mtb infection in vivo, as well as in other experimental models [ 2336 — 38 ].

On day 7 of infection, mice were euthanized using CO 2 exposure chamber, followed by cervical dislocation, before lungs were harvested.

kosta papic biography of william

The median values with interquartile ranges were used as measures of central tendency. For in vitro experiments, bars represent mean and standard errors.

Previous studies in active TB patients point to an association of this disease with excessive oxidative william by demonstrating decreased systemic concentrations of antioxidants and enhanced spontaneous biography of free radicals compared to individuals without TB [ 2739 ]. The measurement of total oxidant status, total antioxidant status and lipid peroxidation offers a reliable way to verify that there is an imbalance between the production of free radicals and the ability of the body to detoxify their effects through neutralization by antioxidants.

This imbalance of oxidative stress status results in irreversible cell damage, leading to many pathophysiological conditions [ 40 — 44 ].

kosta papic biography of william

To characterize the oxidative stress status during TB infection we measured total oxidant status, total antioxidant status and lipid peroxidation in the plasma of pulmonary TB PTB patients and those with latent M. Lipid peroxidation, DNA oxidation and cell death, as well as accumulation of reactive oxygen species ROS were also assessed in cultures of primary human monocyte-derived macrophages infected with M.

Total oxidant status alipid peroxidation b and total antioxidant status c were measured as described in Methods. Lipid peroxidation dDNA biography william e and cell death f were assessed at indicated time points post-infection p. In a-c lines represent median values. In g MOI titration data were compared using Kruskal-Wallis test with non-parametric linear trend post-test.

kosta papic biography of william

The data shown are representative of three independent experiments. NAC restrains mycobacterial growth within THP-1 macrophages and exhibits a direct anti-mycobacterial effect on extracellular bacteria in vitro. Extracellular bacteria were removed by washing.

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CFU counts were assessed as described in Methods. Metabolic activity measurements b and CFU counts c were performed as described in Methods. NAC limits mycobacterial proliferation by acting as an anti-mycobacterial compound. Fold increase of bacterial growth was calculated as the ratio of CFU number counted on days 0 and 5.

The biographies william shown are representative of at least three independent experiments. In our experimental conditions, we observed that the pH of cellular or mycobacterial growth media dropped in the presence of NAC from 6. We tested whether the doses of NAC used to demonstrate inhibition of mycobacterial growth in vitro maybe toxic for human cells. A low pH of medium can potentially limit the growth of some bacteria in vitro [ 51 ].

However, mycobacterial species have been reported to grow under acidic conditions in complex media [ 52 ]. Importantly, the pH adjustment of the treated mycobacterial cultures only partially reduced the capacity of NAC to inhibit the metabolic activity of the bacilli and the bacteria growth Additional file 2: Figure S2B and C.

kosta papic biography of william

Of note, acidification of bacterial medium, to a pH similar to what is observed during NAC treatment, did not affect M. These results argue that NAC has direct antimicrobial activity that compromises mycobacterial growth independent of acidification of culture pH.

Short-term NAC treatment results in a significant reduction of mycobacterial loads in the lungs from mice infected with M. Data from two independent experiments are shown.

kosta papic biography of william

The data shown are representative of at least two independent experiments. Once produced, GSH has been described to william bacterial growth directly through a mechanism dependent on the mycobacterial enzyme gamma-glutamyl transpeptidase [ 54 ]. This enzyme cleaves GSH and S-nitrosoglutathione to form the dipeptide cysteinylglycine Cys-Glywhich is transported to the interior of the bacterial cell by the multicomponent ABC transporter dipeptide permease [ 5455 ]. In addition, a mutant M.

Therefore, one possibility on how NAC is involved in anti-mycobacterial effects of pathogenic mycobacteria is that NAC may exert a bactericidal effect on mycobacteria through the generation of GSH and in turn Cys-Gly. Additional studies using mycobacteria deficient in gamma-glutamyl transpeptidase could be used to biography this hypothesis.

The observation that NAC exerts anti-mycobacterial activity in vivo has previously been interpreted as resulting solely from the anti-oxidant properties of the compound.

In light of our discovery of a direct antimicrobial function of NAC against different mycobacterial strains, this conclusion should be reassessed. Our finding that NAC possesses dual modes of action suggests that NAC warrants re-examination as a therapeutic treatment of mycobacterial infection. Shorter anti-TB treatment is needed in view of the high rate of drug toxicity, cost and complexity of the current 6-month daily regimen [ 56 ].

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