Glutathione and Adaptive Immune Responses against Mycobacterium tuberculosis Infection in Healthy and HIV Infected Individuals

Glutathione (GSH), a tripeptide antioxidant, is essential for cellular homeostasis and plays a vital role in diverse cellular functions. Individuals who are infected with Human immuno deficiency virus (HIV) are known to be susceptible to Mycobacterium tuberculosis (M. tb) infection. We report that by enhancing GSH levels, T-cells are able to inhibit the growth of M. tb inside macrophages. In addition, those GSH-replenished T cell cultures produced increased levels of Interleukin-2 (IL-2), Interleukin-12 (IL-12), and Interferon-gamma (IFN-γ), cytokines, which are known to be crucial for the control of intracellular pathogens. Our study reveals that T lymphocytes that are derived from HIV infected individuals are deficient in GSH, and that this deficiency correlates with decreased levels of Th1 cytokines and enhanced growth of M. tb inside human macrophages

Document classification of Filipino online scam incident text using data mining techniques

The increasing number of online transactions and other internet activities give rise to the proliferation of online scam. The Philippine National Police - Anti Cybercrime Group (PNP-ACG) reported an increasing number of complaints from a double digit figure in 2013 to a triple digit figure in 2017. The challenge of addressing this problem in the Philippines is shared by other developing countries in Southeast Asia and other parts of the world. Since 2013 when the PNP-ACG was established, cybercrime data continue to be accumulated but were not given much attention and significance in research. Previous studies highlight the importance of taking advantage of data mining. However, the absence of empirical studies on cybercrime analytics in the country connotes the lack of exploitation of data mining in facilitating cybercrime investigations. This study exploits Weka text mining tool in order to draw insights by classifying a given online scam dataset. Online scam unstructured data were considered as dataset containing a total of 14, 098 mainly Filipino words. J48 Decision Tree, Naïve Bayes, and Sequential Minimal Optimization were used to build classification models. All these three classifiers or algorithms were compared in terms of performance and prediction accuracy. The results show that J48 achieves the highest accuracy and the lowest error rate, followed by Naïve Bayes and then the SMO classifier. Also, the responses during validation reveal that J48 is preferred over the other classifiers as it easy to understand and apply in cybercrime investigations. This demonstrates how text mining can assist the PNP-ACG in analyzing online scam criminal data as it also highlights the importance of employing data mining tools in the legal and criminal investigation domains in the Philippines. Further work can be carried out in the future using different and a more inclusive cybercrime datasets and other classification techniques in Weka or any other data mining tool, and other data mining tasks such as crime prevention and prediction, clustering, finding leads, trends and patterns of criminal activities, among others. © 2019 IEEE

Intracellular survival of H37Rv inside T cell-monocyte co-cultures.

<p>We determined the intracellular survival of H37Rv inside T cell-monocyte co-cultures from healthy subjects (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0028378#pone-0028378-g003" target="_blank">Figure 3a</a>) and individuals with HIV infection (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0028378#pone-0028378-g003" target="_blank">Figure 3b</a>). Human monocytes were infected with the processed virulent laboratory strain of <i>M. tb</i>, H37Rv at a multiplicity of infection of 10∶1. T cells were purified using nylon wool column. T cells were treated as follows: a) no additives b) NAC (5 mM) c) NAC (10 mM) d) NAC (20 mM) and e) BSO (500 µM) for 24 h. T cells were washed, resuspended in fresh media and then added to the infected monocytes. Infected monocytes-T cell co-cultures were terminated at 1 hour and 5 days post-infection to determine the intracellular survival of H37Rv inside human monocytes. Monocyte lysates were plated on 7H11 medium enriched with ADC to estimate the growth or killing of H37Rv. Results shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0028378#pone-0028378-g003" target="_blank">Figure 3a</a> are averages from five different experiments performed in triplicate. Results shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0028378#pone-0028378-g003" target="_blank">Figure 3b</a> are averages from seven different experiments performed in triplicate.</p

Assay of IL-12, IL-2 and IFN-γin supernatants derived from co-cultures of T cells-<i>M. tb</i>-infected monocytes.

<p>Levels of IL-12, IL-2 and IFN-γ were assayed in supernatants derived from co-cultures of T cells-<i>M. tb</i>-infected monocytes from healthy individuals (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0028378#pone-0028378-g004" target="_blank">Figure 4a, b and c</a>) and individuals with HIV infection (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0028378#pone-0028378-g004" target="_blank">Figure 4d and e</a>) T cells were purified using nylon wool column. T cells were treated as follows: 1) no additives 2) NAC (5 mM) 3) NAC (10 mM) 4) NAC (20 mM) and 5) BSO (500 µM) for 24 h. Following incubation with stimulants, T cells were washed, re-suspended in fresh RPMI containing human serum without any stimulants and then added to the infected monocytes. Supernatants were collected from co-cultures of H37Rv-infected monocytes-T cells at 5 days post-infection were filtered and assayed for the levels of IL-12, IL-2 and IFN-γ using assay kits from eBioscience. Data in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0028378#pone-0028378-g004" target="_blank">Figures 4a, b and c</a> represent means±SE from four different healthy individuals. Data in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0028378#pone-0028378-g004" target="_blank">Figures 4d and e</a> represent means±SE from six different individuals with HIV infection.</p

Assay of free radicals and TNF-α in plasma samples from healthy and HIV positive individuals.

<p>Plasma samples separated from blood of healthy volunteers and HIV-infected individuals were used for measurement of free radicals (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0028378#pone-0028378-g002" target="_blank">Figure 2a</a>) and TNF-α (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0028378#pone-0028378-g002" target="_blank">Figure 2b</a>). Free radical levels in plasma samples derived from healthy subjects and individuals with HIV infection was determined by measuring the levels of MDA using a colorimetric assay kit from Cayman. Levels of TNF-α in the plasma samples were determined by ELISA using assay kits procured from eBioscience. Results in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0028378#pone-0028378-g002" target="_blank">Figure 2</a> are averages of data collected from thirteen healthy subjects and thirteen individuals with HIV infection.</p

Assay of IL-12, IL-2, IFN-γ and IL-10 in plasma samples from healthy and HIV positive individuals.

<p>Plasma samples separated from blood of healthy volunteers and HIV-infected individuals were used for measurement of IL-12 (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0028378#pone-0028378-g005" target="_blank">Figure 5a</a>), IL-2 (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0028378#pone-0028378-g005" target="_blank">Figure 5b</a>), IFN-γ (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0028378#pone-0028378-g005" target="_blank">Figure 5c</a>) and IL-10 (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0028378#pone-0028378-g005" target="_blank">Figure 5d</a>) by ELISA using assay kits from eBioscience. Results in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0028378#pone-0028378-g005" target="_blank">Figure 5</a> are averages from data collected from thirteen healthy subjects and thirteen individuals with HIV infection.</p

Model describing the mechanism by which GSH-enhanced T cells control <i>M. tb</i> infection in macrophages.

<p>Model describing the mechanism by which GSH-enhanced T cells control <i>M. tb</i> infection in macrophages.</p

Assay of GSH levels in T cells isolated from healthy and HIV positive individuals.

<p>Intracellular levels of GSH in freshly isolated T cells from healthy volunteers and HIV-infected individuals was determined by spectrophotometry, using an assay kit from Calbiochem (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0028378#pone-0028378-g001" target="_blank">Figure 1a</a>). T cells (2×10⁵/well) purified from PBMCs using nylon wool columns were pelleted by centrifugation and an equal volume of ice cold 5% MPA was added to the pellet. Supernatants were collected after centrifugation and analyzed for total GSH using an assay kit from Calbiochem, as per manufacturer’s instructions. Total GSH in the samples were normalized with protein. Proteins in the samples were estimated by Bradford’s method using Bio-Rad reagent. We also tested the effects of NAC and BSO in increasing and decreasing the intracellular levels of GSH, respectively in T cells isolated from both healthy subjects and individuals with HIV infection (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0028378#pone-0028378-g001" target="_blank">Figure 1b</a>). T cells (2×10⁵/well) isolated from healthy and HIV positive subjects were treated as follows: mock treatment, treatment with NAC (10 mM) and treatment with BSO (500 µM). Following overnight incubation, T cells were pelleted and used for GSH measurement as per manufacturer’s instructions. Results shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0028378#pone-0028378-g001" target="_blank">Figure 1b</a> are averages from experiments performed using T cells isolated from three healthy individuals and eight individuals with HIV infection. * represent significant difference in the levels of GSH between untreated T cells derived from healthy subjects versus untreated T cells derived from individuals with HIV infection. **denotes significant difference in the levels of GSH between untreated T cells versus NAC-treated T cells from HIV positive subjects. *** denotes statistically significant difference in the levels of GSH between NAC-treated T cells and BSO-treated T cells from individuals with HIV infection.</p