Interaction between Cfd1 and Nbp35 proteins involved in cytosolic FeS cluster assembly machinery deciphers a stable complexation in Leishmania donovani.

Leishmania donovani is the causative unicellular parasite for visceral leishmaniasis (VL); and FeS proteins are likely to be very essential for their survival and viability. Cytosolic FeS cluster assembly (CIA) machinery is one of the four systems for the biosynthesis and transfer of FeS clusters among eukaryotes; Cfd1 and Nbp35 are the scaffold components for cytosolic FeS cluster biogenesis. We investigated the role of CIA machinery components and purified Cfd1 and Nbp35 proteins of L. donovani. We also investigated the interactive nature between LdCfd1 and LdNbp35 proteins by in silico analysis, in vitro co-purification, pull down assays along with in vivo immuno-precipitation; which inferred that both LdCfd1 and LdNbp35 proteins are interacting with each other. Thus, our collective data revealed the interaction between these two proteins which forms a stable complex that can be attributed to the cellular process of FeS clusters biogenesis, and transfer to target apo-proteins of L. donovani. The expression of Cfd1 and Nbp35 proteins in Amp B resistant parasites is up-regulated leading to increased amount of FeS proteins. Hence, it favors increased tolerance towards ROS level, which helps parasites survival under drug pressure contributing in Amphotericin B resistance.

Improving Indoor Air Ventilation by a Ceiling Fan to Mitigate Aerosols Transmission.

Improving air flow and ventilation in an indoor environment is central to mitigating the airborne transmission of aerosols. Examples include, COVID-19 or similar diseases that transmit by airborne aerosols or respiratory droplets. While there are standard guidelines for enhancing the ventilation of space, the effect of a ceiling fan on the ventilation has not been explored. Such an intervention could be critical, especially in a resource-limited setting. In the present work, we numerically study the effect of a rotating ceiling fan on indoor air ventilation using computational fluid dynamics (CFD) simulations. In particular, we employ RANS turbulence model and compare the computed flow fields for a stationary and rotating fan in an office room with a door and window. While a re-circulation zone spans the whole space for the stationary fan, stronger re-circulation zones and small stagnation zones appear in the flow-field inside the room for the case of a rotating fan. The re-circulation zones help bring in fresh air through the window and remove stale air through the door, thereby improving the ventilation rate by one order of magnitude. We briefly discuss the chances of infection by aerosols via flow-fields corresponding to stationary and rotating fans.

Unique thiol metabolism in trypanosomatids: Redox homeostasis and drug resistance.

Trypanosomatids are mainly responsible for heterogeneous parasitic diseases: Leishmaniasis, Sleeping sickness, and Chagas disease and control of these diseases implicates serious challenges due to the emergence of drug resistance. Redox-active biomolecules are the endogenous substances in organisms, which play important role in the regulation of redox homeostasis. The redox-active substances like glutathione, trypanothione, cysteine, cysteine persulfides, etc., and other inorganic intermediates (hydrogen peroxide, nitric oxide) are very useful as defence mechanism. In the present review, the suitability of trypanothione and other essential thiol molecules of trypanosomatids as drug targets are described in Leishmania and Trypanosoma. We have explored the role of tryparedoxin, tryparedoxin peroxidase, ascorbate peroxidase, superoxide dismutase, and glutaredoxins in the anti-oxidant mechanism and drug resistance. Up-regulation of some proteins in trypanothione metabolism helps the parasites in survival against drug pressure (sodium stibogluconate, Amphotericin B, etc.) and oxidative stress. These molecules accept electrons from the reduced trypanothione and donate their electrons to other proteins, and these proteins reduce toxic molecules, neutralize reactive oxygen, or nitrogen species; and help parasites to cope with oxidative stress. Thus, a better understanding of the role of these molecules in drug resistance and redox homeostasis will help to target metabolic pathway proteins to combat Leishmaniasis and trypanosomiases.

Effect of co-flow on fluid dynamics of a cough jet with implications in spread of COVID-19.

We discuss the temporal evolution of a cough jet of an infected subject in the context of the spread of COVID-19. Computations were carried out using large eddy simulation, and, in particular, the effect of the co-flow (5% and 10% of maximum cough velocity) on the evolution of the jet was quantified. The Reynolds number (Re) of the cough jet, based on the mouth opening diameter (D) and the average cough velocity, is 13 002. The time-varying inlet velocity profile of the cough jet is represented as a combination of gamma-probability-distribution functions. Simulations reveal the detailed structure of cough jet with and without a co-flow for the first time, to the best of our knowledge. The cough jet temporal evolution is similar to that of a continuous free-jet and follows the same routes of instability, as documented for a free-jet. The convection velocity of the cough jet decays with time and distance, following a power-law variation. The cough jet is observed to travel a distance of approximately 1.1 m in half a second. However, in the presence of 10% co-flow, the cough jet travels faster and covers the similar distance in just 0.33 s. Therefore, in the presence of a co-flow, the probability of transmission of COVID-19 by airborne droplets and droplet nuclei increases, since they can travel a larger distance. The cough jet without the co-flow corresponds to a larger volume content compared to that with the co-flow and spreads more within the same range of distance. These simulations are significant as they help to reveal the intricate structure of the cough jet and show that the presence of a co-flow can significantly augment the risk of infection of COVID-19.

The HIV - 1 protease inhibitor Amprenavir targets Leishmania donovani topoisomerase I and induces oxidative stress-mediated programmed cell death.

The global prevalence of HIV is a major challenge for the control of visceral leishmaniasis. Although the effectiveness and usefulness of amprenavir (APV) are well studied in anti-retroviral regimens, very little is known on HIV/VL-co-infected patients. In the present study, we report for the first time the protective efficacy of APV against visceral leishmaniasis by inhibition of DNA Topoisomerase I (LdTOP1LS) and APV-induced downstream pathway in programmed cell death (PCD). During the early phase of activation, reactive oxygen species (ROS) is increased inside the cells, which causes subsequent elevation of lipid peroxidation. Endogenous ROS formation and lipid peroxidation cause eventual depolarization of mitochondrial membrane potential (ΔΨm). Furthermore, the release of cytochrome c and activation of CED3/CPP32 group of proteases lead to the formation of oxidative DNA lesions followed by DNA fragmentation. The promising in vitro and ex vivo results promoted to substantiate further by in vivo animal experiment, which showed a significant reduction of splenic and hepatic parasites burden compared to infected controls. Interestingly, APV selectively targets LdTOPILS and does not inhibit the catalytic activity of human topoisomerase I (hTopI). Moreover, based on the cytotoxicity test APV is not toxic for host macrophage cells, which is correlated with non-responsiveness of inhibition of catalytic activity of hTopI. Taken together, this study provides the opportunity for discovering and evaluating newer potential molecular therapeutic targets for drug designing. The present study might be exploited in future as important therapeutics, which will be useful for treatment of VL as well as HIV-VL co-infection.

Detection and functional characterization of sigma class GST in Phlebotomus argentipes and its role in stress tolerance and DDT resistance.

Several Glutathione S-transferases (GSTs) enzymes, in insects, have previously been implicated in resistance developed against DDT and other insecticides. The GST enzyme particularly sigma class have important physiological role in detoxification of lipid peroxidation by-products in insects. Phlebotomus argentipes has been intensely exposed to DDT over years due to Indoor Residual Spray (IRS) programme for Kala-azar elimination in Bihar, India. However, in P. argentipes, role of GSTs in DDT resistance have not been elucidated. Here, sigma class GST of P. argentipes (Parg-GSTσ) was successfully cloned, expressed and purified by affinity chromatography. The recombinant Parg-GSTσ was found to be highly active towards cumene hydroperoxide and 4-HNE having specific activity 92.47 & 203.92 µM/min/mg of protein, respectively and exhibited low activity towards universal substrate CDNB i.e., 8.75 µM/min/mg of protein. RT-PCR and immunoblot analysis showed at least 2 and 1.8 fold overexpression of Parg-GSTσ in the single exposed and non exposed DDT resistant P. argentipes as compared to susceptible, implicating Parg-GSTσ also involved in DDT resistance probably by imparting enhanced stress tolerance. The DDT, H2O2 and temperature induction assays demonstrated stress-dependent induction of Parg-GSTσ expression indicating its important role in oxidative stress redressal.