ABSTRACT
The presence of antioxidant enzymes in helminth parasites is well known. These enzymes help the parasites to survive in their hosts by detoxifying host-generated reactive oxygen species (ROS). The literature survey reveals that most of the studies related to antioxidant enzymes in helminth parasites are restricted to the adult stage while the larval stages are neglected. The present investigation is designed to evaluate the level of antioxidant enzymes in the adult and larval stages of rumen-infecting paramphistome parasites, Gastrothylax crumenifer. The larval stages include 0-day eggs, 4-day eggs, and eggs containing mature miracidia, cercariae, and metacercariae. Antioxidant enzyme assays were performed using standard assay protocols. Our findings revealed an increasing pattern in the level of Glutathione-S-Transferase (GST), Superoxide Dismutase (SOD), Glutathione Reductase (GR), and Glutathione Peroxidase (GPx) antioxidant enzymes during the development from 0-day eggs to the adult stage. Overall analysis shows that adult worms have higher antioxidant enzyme activity as compared to the larval stages, indicating that adult flukes are more adapted to oxidative stress. It can be concluded that the miracidia, cercarial, and metacercarial developmental stages of G. crumenifer possess a considerable level of antioxidant enzymes suitable to overcome the oxidative stress encountered during the development and help them in the completion of the life cycle and survival in the definitive host.
ABSTRACT
Due to unique inherent catalytic characteristics of different size, shape and surface functionalized gold nanoparticles, their potential applications, are being explored in various fields such as drug delivery, biosensor, diagnosis and theranostics. However conventional process for synthesis of these metallic nanoparticles utilizes toxic reagents as reducing agents, additional capping agent for stability as well as surface functionalization for drug delivery purposes. Hence, in this work suitability of gum Ghatti for reducing, capping and surface functionalization during the synthesis of stable Gold nanoparticles were duly explored. Role and impact of key process variables i.e. volume of chloroauric acid solution, gum solution and temperature at their respective three different levels, as well as mechanism of formation of optimized gold nanoparticles were also investigated using Box- Behnken design. These novel synthesized optimized Gold nanoparticles were further characterized by UV spectrophotometer for its surface plasmon resonance (SPR) at around â¼530nm, dynamic light scattering (DLS) for its hydrodynamic size (112.5nm), PDI (0.222) and zeta potential (-21.3mV) while, transmission electron microscopy (TEM) further revealed surface geometry of these nanoparticles being spherical in shape.
Subject(s)
Gold/chemistry , Metal Nanoparticles/chemistry , Nanotechnology/methods , Plant Gums/chemistry , Statistics as Topic , Chemistry Techniques, Synthetic , Green Chemistry Technology , Models, StatisticalABSTRACT
AIM: Aim of the present study was to carry out the partial purification and biochemical characterization of glutathione S-transferase (GST) from the somatic tissue of ruminal amphistome parasite, Gastrothylax crumenifer (Gc) infecting Indian water buffalo (Bubalus bubalis). MATERIALS AND METHODS: The crude somatic homogenate of Gc was subjected to progressive ammonium sulfate precipitation followed by size exclusion chromatography in a Sephacryl S 100-HR column. The partially purified GST was assayed spectrophotometrically, and the corresponding enzyme activity was also recorded in polyacrylamide gel. GST isolated from the amphistome parasite was also exposed to variable changes in temperature and the pH gradient of the assay mixture. RESULTS: The precipitated amphistome GST molecules showed maximum activity in the sixth elution fraction. The GST subunit appeared as a single band in the reducing polyacrylamide gel electrophoresis with an apparent molecular weight of 26 kDa. The GST proteins were found to be fairly stable up to 37°C, beyond this the activity got heavily impaired. Further, the GST obtained showed a pH optima of 7.5. CONCLUSION: Present findings showed that GST from Gc could be conveniently purified using gel filtration chromatography. The purified enzyme showed maximum stability and activity at 4°C.
