Acanthamoeba spp. cause a corneal infection, Acanthamoeba keratitis (AK), and a cerebral infection, granulomatous amoebic encephalitis (GAE). Though aggressive chemotherapy has been able to kill the active trophozoite form of Acanthamoeba, the encysted form of this parasite has remained problematic to resist physiological concentrations of drugs. The emergence of encysted amoeba into active trophozoite form poses a challenge to eradicate this parasite. Acanthamoeba trophozoites have active metabolic machinery that furnishes energy in the form of ATPs by subjecting carbohydrates and lipids to undergo pathways including glycolysis and beta-oxidation of free fatty acids, respectively. However, very little is known about the metabolic preferences and dependencies of an encysted trophozoite on minerals or potential nutrients that it consumes to live in an encysted state. Here, we investigate the metabolic and nutrient preferences of the encysted trophozoite of Acanthamoeba castellanii and the possibility to target them by drugs that act on calcium ion dependencies of the encysted amoeba. The experimental assays, immunostaining coupled with bioinformatics tools show that the encysted Acanthamoeba uses diverse nutrient pathways to obtain energy in the quiescent encysted state. These findings highlight potential pathways that can be targeted in eradicating amoebae cysts successfully.
The generation of power and fuel sustainability that contributes to a cleaner output of exhaust gases is one of the most important objectives the world seeks. In this paper, oxyhydrogen gas is used to retrofit into a two-stroke engine. The water was electrolysed and generated a mixture of oxygen (O2) and hydrogen (H2) or known as oxyhydrogen (HHO) gas via an electrolytic dry cell generator. The HHO was retrofitted experimentally to investigate the engine emissions and exhaust gas temperature from a 1.5 kW gasoline engine. The engine was tested with different power ratings (84-720 W) to investigate the performance and emissions of the engine using gasoline followed by the addition of HHO. The emissions of CO and NOx were measured with different amounts of HHO added. The exhaust temperature was calculated as one of the variables to be considered in relation to pollution. The air-fuel ratios are varied from 12 to 20% in the experiment. The most appropriate air-fuel ratio needed to start the generator with the most environmentally friendly gas emission was analysed. The results showed that the addition of HHO to the engine is successful in reducing fuel consumption up to 8.9%. A higher percentage of HHO added also has improved the emissions and reduced exhaust gas temperature. In this study, the highest quantity of HHO added at 0.15% of the volume fraction reduced CO gas emission by up to 9.41%, NOx gas up to 4.31%, and exhaust gas temperature by up to 2.02%. Generally, adding oxyhydrogen gas has significantly reduced the emissions, and exhaust temperature and provided an eco-friendly environment.