Methanol Extract of Polyopes lancifolius Inhibits the Expression of Pro-inflammatory Mediators in LPSstimulated BV2 Microglia Cells via Downregulation of the NF-&#954B Pathway

Purpose: This study is aimed at identifying the anti-inflammatory mechanisms of a methanol extract of Polyopes lancifolius (MEPL) in lipopolysaccharide (LPS)-stimulated BV2 microglia cells.Methods: The expression of mRNA and protein were investigated RT-PCR and western blot analyses in LPS-stimulated BV2 microglial cells. The level of nitric oxide (NO) production was analyzed using Griess reaction. The release of prostaglandin E2 (PGE2) and tumor necrosis factor-α (TNF-α) were determined using sandwich ELISA. NF-κB activation was detected using EMSA methods.Results: MEPL significantly suppressed NO production in LPS-stimulated BV2 cells without any cytotoxicity. The results also indicate that MEPL decreased the production of PGE2 and TNF-α in LPSstimulated BV2 cells. Furthermore, pretreatment with MEPL resulted in a downregulation of LPSinduced mRNA and protein expression of inducible NO synthase (iNOS), cyclooxygenase-2 (COX-2) and TNF-α. Investigation of the effect of MEPL on nuclear factor-κB (NF-κB) activity, which is a potential transcriptional factor for regulating inflammatory genes such as iNOS, COX-2 and TNF-α, showed that MEPL substantially inhibited the LPS-induced DNA-binding activity of NF-κB. MEPL also suppressed the LPS-induced degradation and phosphorylation of I&kappaBα, and it consequently blocked p65 translocation from the cytosol to the nucleus.Conclusion: These data show that MEPL may regulate LPS-induced NO, PGE2, and TNF-α production by suppressing NF-κB activity.Keywords: Polyopes lancifolius, Nitric oxide, Prostaglandin E2, Tumor necrosis factor-α, Nuclear factor-κ

Digitalization-A Strategic Approach to Promote the Hospitality Industry in Order to Increase Tourism in Sri Lanka with Monetary Gain

The purpose of this research was to empower hospitality management and because of that increase tourism attraction. In 2018 direct contribution to GDP was 4.9% It was decrease to 4.3% after the 2019 Easter attack. In 2020 this hugely decreased to 0.8% according to the Tourism Authority of Sri Lanka. Covid 19 impact was affected by that. This study aims at explaining in detail how technology can use to improve the tourism sector. Therefore, the digitalized hotel management system was implemented as a solution for that

Commentary - Better lung health for all: A British Thoracic Society and Pan African Thoracic Society initiative

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Methanol Extract of Myelophycus caespitosus Inhibits the Inflammatory Response in Lipopolysaccharidestimulated BV2 Microglial Cells by Downregulating NF-kB via Inhibition of the Akt Signaling Pathway

Purpose: To determine whether the methanol extract of Myelophycus caespitosus (MEMC) downregulates the expression of pro-inflammatory mediators in lipopolysaccharide (LPS)-stimulated BV2 microglial cells.Methods: Reverse transcription-polymerase chain reaction (RT-PCR) together with Western blot analysis was used to evaluate the expression of pro-inflammatory mediators such as nitric oxide (NO) and prostaglandin E2(PGE2) as well as their regulatory genes such as inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2), in LPS-stimulated BV2 microglial cells. The level of NO production was analyzed using Griess reaction.The release of PGE2 was determined using sandwich enzyme-linked immunosorbent assay. The DNA-binding activity of nuclear factor-êB (NF-êB) was measured by electrophoretic mobility shift assay.Results: MEMC inhibited LPS-induced pro-inflammatory mediators, NO and PGE2, as well as their respective genes, iNOS and COX-2, at both protein and mRNA levels, without any significant cytotoxicity. Treatment withMEMC also substantially reduced the LPS-induced DNA-binding activity of NF-êB and nuclear translocation of NF-êB subunits p65 and p50 via the inhibition of IêBá phosphorylation and degradation. MEMC promoteddephosphorylation of Akt that subsequently suppressed the DNA-binding activity of NF-êB in LPS-stimulated BV2 microglial cells.Conclusion: Collectively, these data suggest that MEMC attenuates expression of pro-inflammatory mediators such as NO and PGE2 by suppression of their regulatory genes through the inhibition of Aktmediated NF-êB activity.Keywords: Myelophycus caespitosus, Nitric oxide, Prostaglandin E2, Nuclear factor-êB

Histoplasmosis in Sri Lanka - a masquerader in a strange land: A Case Report

We present a patient with an oral mucosal ulcer who was diagnosed with disseminated histoplasmosis. This fungal infection is endemic in the United States, and thus may be not considered in the differential diagnosis of oral ulcers in Sri Lanka. Furthermore, it may mimic many common diseases found in Sri Lanka. It is therefore important to be aware of this entity, since it is potentially curable if diagnosed and treated early.</p

The burden of non-TB lung disease presenting to TB clinics in The Gambia: preliminary data in the Xpert® MTB/Rif era

In some low and middle-income countries, 10-20% of patients presenting with a persistent cough have tuberculosis (TB). Once TB is excluded, health service provision for alternative diagnoses is limited. We prospectively studied patients with two Xpert-negative sputum results presenting to a TB clinic in The Gambia. Of 239 patients, 108 did not have TB; 65/102 (6 were lost to follow-up) had alternative diagnoses, 24.6% of which were non-respiratory; 37/102 had no diagnosis, 27.0% of whom were HIV-1-positive; 37.8% had a history of TB and 24.3% smoked. We highlight the need for general health service integration with TB platforms and exploration of non-TB patients with chronic respiratory symptoms

Quantifying the global number of tuberculosis survivors: a modelling study

Background People who survive tuberculosis face clinical and societal consequences after recovery, including increased risks of recurrent tuberculosis, premature death, reduced lung function, and ongoing stigma. To describe the size of this issue, we aimed to estimate the number of individuals who developed first-episode tuberculosis between 1980 and 2019, the number who survived to 2020, and the number who have been treated within the past 5 years or 2 years. Methods In this modelling study, we estimated the number of people who survived treated tuberculosis using country-level WHO data on tuberculosis case notifications, excluding those who died during treatment. We estimated the number of individuals surviving untreated tuberculosis using the difference between WHO country-level incidence estimates and notifications, applying published age-stratified and HIV-stratified case fatality ratios. To estimate survival with time, post-tuberculosis life tables were developed for each country-year by use of UN World Population Prospects 2019 mortality rates and published post-tuberculosis mortality hazard ratios. Findings Between 1980 and 2019, we estimate that 363 million people (95% uncertainty interval [UI] 287 million–438 million) developed tuberculosis, of whom 172 million (169 million–174 million) were treated. Individuals who developed tuberculosis between 1980 and 2019 had lived 3480 million life-years (95% UI 3040 million–3920 million) after tuberculosis by 2020, with survivors younger than 15 years at the time of tuberculosis development contributing 12% (95% UI 7–17) of these life-years. We estimate that 155 million tuberculosis survivors (95% UI 138 million–171 million) were alive in 2020, the largest proportion (47% [37–57]) of whom were in the WHO South-East Asia region. Of the tuberculosis survivors who were alive in 2020, we estimate that 18% (95% UI 16–20) were treated in the past 5 years and 8% (7–9) were treated in the past 2 years. Interpretation The number of tuberculosis survivors alive in 2020 is more than ten times the estimated annual tuberculosis incidence. Interventions to alleviate respiratory morbidity, screen for and prevent recurrent tuberculosis, and reduce stigma should be immediately prioritised for recently treated tuberculosis survivors. Funding UK Medical Research Council, the UK Department for International Development, the National Institute for Health Research, and the European and Developing Countries Clinical Trials Partnership

Plasmodium Infection Is Associated with Impaired Hepatic Dimethylarginine Dimethylaminohydrolase Activity and Disruption of Nitric Oxide Synthase Inhibitor/Substrate Homeostasis.

Inhibition of nitric oxide (NO) signaling may contribute to pathological activation of the vascular endothelium during severe malaria infection. Dimethylarginine dimethylaminohydrolase (DDAH) regulates endothelial NO synthesis by maintaining homeostasis between asymmetric dimethylarginine (ADMA), an endogenous NO synthase (NOS) inhibitor, and arginine, the NOS substrate. We carried out a community-based case-control study of Gambian children to determine whether ADMA and arginine homeostasis is disrupted during severe or uncomplicated malaria infections. Circulating plasma levels of ADMA and arginine were determined at initial presentation and 28 days later. Plasma ADMA/arginine ratios were elevated in children with acute severe malaria compared to 28-day follow-up values and compared to children with uncomplicated malaria or healthy children (p<0.0001 for each comparison). To test the hypothesis that DDAH1 is inactivated during Plasmodium infection, we examined DDAH1 in a mouse model of severe malaria. Plasmodium berghei ANKA infection inactivated hepatic DDAH1 via a post-transcriptional mechanism as evidenced by stable mRNA transcript number, decreased DDAH1 protein concentration, decreased enzyme activity, elevated tissue ADMA, elevated ADMA/arginine ratio in plasma, and decreased whole blood nitrite concentration. Loss of hepatic DDAH1 activity and disruption of ADMA/arginine homeostasis may contribute to severe malaria pathogenesis by inhibiting NO synthesis

Monitoring anti-tuberculosis treatment response using analysis of whole blood Mycobacterium tuberculosis specific T cell activation and functional markers

Background: Blood-based biomarkers have been proposed as an alternative to current sputum-based treatment monitoring methods in active tuberculosis (ATB). The aim of this study was to validate previously described phenotypic, activation, and cytokine markers of treatment response in a West African cohort. Methods: Whole blood immune responses to Mycobacterium tuberculosis ESAT-6/CFP-10 (EC) and purified protein derivative (PPD) were measured in twenty adults at baseline and after 2 months of standard TB treatment. Patients were classified as fast or slow responders based on a negative or positive sputum culture result at 2 months, respectively. Cellular expression of activation markers (CD38, HLA-DR), memory markers (CD27), and functional intracellular cytokine and proliferation (IFN-γ, Ki-67, TNF-α) markers were measured using multi-color flow cytometry. Results: There was a significant increase in the proportion of CD4+CD27+ cells expressing CD38 and HLA-DR following EC stimulation at 2 months compared to baseline (p = 0.0328 and p = 0.0400, respectively). Following PPD stimulation, slow treatment responders had a significantly higher proportion of CD8+CD27–IFN-γ+ (p = 0.0105) and CD4+CD27+HLA-DR+CD38+ (p = 0.0077) T cells than fast responders at baseline. Receiver operating curve analysis of these subsets resulted in 80% sensitivity and 70 and 100% specificity, respectively (AUC of 0.82, p = 0.0156 and 0.84, p = 0.0102). Conclusion: Our pilot data show reductions in expression of T cell activation markers were seen with treatment, but this was not associated with fast or slow sputum conversion at 2 months. However, baseline proportions of activated T cell subsets are potentially predictive of the subsequent speed of response to treatment

Distinct roles for FOXP3(+) and FOXP3(-) CD4(+) T cells in regulating cellular immunity to uncomplicated and severe plasmodium falciparum Malaria

Failure to establish an appropriate balance between pro- and anti-inflammatory immune responses is believed to contribute to pathogenesis of severe malaria. To determine whether this balance is maintained by classical regulatory T cells (CD4+ FOXP3+ CD127−/low; Tregs) we compared cellular responses between Gambian children (n = 124) with severe Plasmodium falciparum malaria or uncomplicated malaria infections. Although no significant differences in Treg numbers or function were observed between the groups, Treg activity during acute disease was inversely correlated with malaria-specific memory responses detectable 28 days later. Thus, while Tregs may not regulate acute malarial inflammation, they may limit memory responses to levels that subsequently facilitate parasite clearance without causing immunopathology. Importantly, we identified a population of FOXP3−, CD45RO+ CD4+ T cells which coproduce IL-10 and IFN-γ. These cells are more prevalent in children with uncomplicated malaria than in those with severe disease, suggesting that they may be the regulators of acute malarial inflammation