Acanthamoeba is an opportunistic protist pathogen that is responsible for serious human and animal infection. Being one of the most frequently isolated protists from the environment, it is likely that it readily encounters microaerophilic environments. For respiration under anaerobic or low oxygen conditions in several amitochondriate protists, decarboxylation of pyruvate is catalyzed by pyruvate ferredoxin oxidoreductase instead of pyruvate dehydrogenase. In support, Nitazoxanide, an inhibitor of pyruvate ferredoxin oxidoreductase, is effective and non-mutagenic clinically against a range of amitochondriate protists, Giardia intestinalis, Entamoeba histolytica and Trichomonas vaginalis. The overall aim of the present study was to determine in vitro efficacy of Nitazoxanide against Acanthamoeba castellanii. At micromolar concentrations, the findings revealed that Nitazoxanide neither affected A. castellanii growth or viability nor amoeba-mediated host cell monolayer damage in vitro or extracellular proteolytic activities. Similarly, microaerophilic conditions alone had no significant effects. In contrast, microaerophilic conditions together with Nitazoxanide showed amoebicidal effects and inhibited A. castellanii-mediated host cell monolayer damage as well as extracellular proteases. Using encystation assays, it was observed that Nitazoxanide inhibited trophozoite transformation into cysts both under aerophilic and microaerophilic conditions. Furthermore, pre-treatment of cysts with Nitazoxanide inhibited A. castellanii excystation. These findings are important in the identification of potential targets that could be useful against parasite-specific respiration as well as to understand the basic biology of the life cycle of Acanthamoeba.
Eight novel N'-(2-oxoindolin-3-ylidene)-2-propylpentane hydrazide-hydrazone derivatives 4a-h were synthesized and fully characterized by IR, NMR ((1)H-NMR and (13)C-NMR), elemental analysis, and X-ray crystallography. The cyto-toxicity and in vitro anti-cancer evaluation of the prepared compounds have been assessed against two different human tumour cell lines including human liver (HepG2) and leukaemia (Jurkat), as well as in normal cell lines derived from human embryonic kidney (HEK293) using MTT assay. The compounds 3e, 3f, 4a, 4c, and 4e revealed promising anti-cancer activities in tested human tumour cells lines (IC50 values between 3 and 7 μM) as compared to the known anti-cancer drug 5-Fluorouracil (IC50 32-50 μM). Among the tested compounds, 4a showed specificity against leukaemia (Jurkat) cells, with an IC50 value of 3.14 μM, but this compound was inactive in liver cancer and normal cell lines.
The majority of influenza vaccines are manufactured using embryonated hens' eggs. The potential occurrence of a pandemic outbreak of avian influenza might reduce or even eliminate the supply of eggs, leaving the human population at risk. Also, the egg-based production technology is intrinsically cumbersome and not easily scalable to provide a rapid worldwide supply of vaccine. In this communication, the production of a cell culture (Madin-Darby canine kidney (MDCK)) derived live attenuated influenza vaccine (LAIV) in a fully disposable platform process using a novel Single Use Bioreactor (SUB) is presented. The cell culture and virus infection was maintained in a disposable stirred tank reactor with PID control of pH, DO, agitation, and temperature, similar to traditional glass or stainless steel bioreactors. The application of this technology was tested using MDCK cells grown on microcarriers in proprietary serum free medium and infection with 2006/2007 seasonal LAIV strains at 25-30 L scale. The MDCK cell growth was optimal at the agitation rate of 100 rpm. Optimization of this parameter allowed the cells to grow at a rate similar to that achieved in the conventional 3 L glass stirred tank bioreactors. Influenza vaccine virus strains, A/New Caledonia/20/99 (H1N1 strain), A/Wisconsin/67/05 (H3N2 strain), and B/Malaysia/2506/04 (B strain) were all successfully produced in SUB with peak virus titers > or =8.6 log(10) FFU/mL. This result demonstrated that more than 1 million doses of vaccine can be produced through one single run of a small bioreactor at the scale of 30 L and thus provided an alternative to the current vaccine production platform with fast turn-around and low upfront facility investment, features that are particularly useful for emerging and developing countries and clinical trial material production.
Vector-host infectious diseases remain a challenging issue and cause millions of deaths each year globally. In such outbreaks, many countries especially developing or underdevelopment faces a situation where the number of infected individuals is getting larger and the medical facilities are limited. In this paper, we construct an epidemic model to explore the transmission dynamics of vector-borne diseases with nonlinear saturated incidence rate and saturated treatment function. This type of incidence rate, as well as the saturated treatment function, is also known as the Holling type II form and describes the effect of delayed treatment. Initially, we formulate a mathematical model and then present the basic analysis of the model including the positivity and boundedness of the solution. The threshold quantity R 0 is presented and the stability analysis of the system is carried out for the model equilibria. The global stability results are shown using the Lyapunov function of Goh-Voltera type. The existence of backward bifurcation is discussed using the central manifold theory. Further, the global sensitivity analysis of the model is carried out using the Latin Hypercube sampling and the partial rank correlation coefficient techniques. Moreover, an optimal control problem is formulated and the necessary optimality conditions are investigated in order to eradicate the disease in a community. Four strategies are presented by choosing different set of controls combination for the disease minimization. Finally, the numerical simulations of each strategy are depicted to demonstrate the importance of suggesting control interventions on the disease dynamics and eradication.
Herein, a novel and environmentally benign solid catalyst was fabricated by grafting WO3 active species onto the ZnCo2O4@CeO2 support for efficient levulinic acid production from corncob waste biomass. The morphological, compositional, and textural properties of the designed catalyst were investigated using different characterization techniques to identify suitable catalyst formulation with enhanced catalytic activity and stability. The results demonstrated that WO3 active species were successfully loaded with uniform distribution onto the support to develop a robust catalyst with both acidic and basic sites. The experimental investigation showed that among the catalysts, WO3(10 wt %)/ZnCo2O4@CeO2 exhibited the best catalytic activity, providing a maximum levulinic acid yield of 78.49% at the optimal conditions of 6 wt % catalyst dosage, reaction temperature of 180 °C, and reaction time of 200 min. The presence of an optimum number of both acid and base active sites on the catalyst surface could lead to the highest catalytic activity of the synthesized catalyst. Finally, the reusability investigation indicated that the synthesized catalyst possessed sufficient recyclability of up to four times for the levulinic acid production from the selected biomass with negligible drop in the catalytic activity.
Chimeric antigen receptor (CAR) T therapy has shown remarkable success in discovering novel CAR-T cell products for treating malignancies. Despite of successful results from clinical trials, CAR-T cell therapy is ineffective for long-term disease progression. Numerous challenges of CAR-T cell immunotherapy such as cell dysfunction, cytokine-related toxicities, TGF-β resistance, GvHD risks, antigen escape, restricted trafficking, and tumor cell infiltration still exist that hamper the safety and efficacy of CAR-T cells for malignancies. The accumulated data revealed that these challenges could be overcome with the advanced CRISPR genome editing technology, which is the most promising tool to knockout TRAC and HLA genes, inhibiting the effects of dominant negative receptors (PD-1, TGF-β, and B2M), lowering the risks of cytokine release syndrome (CRS), and regulating CAR-T cell function in the tumor microenvironment (TME). CRISPR technology employs DSB-free genome editing methods that robustly allow efficient and controllable genetic modification. The present review explored the innovative aspects of CRISPR/Cas9 technology for developing next-generation/universal allogeneic CAR-T cells. The present manuscript addressed the ongoing status of clinical trials of CRISPR/Cas9-engineered CAR-T cells against cancer and pointed out the off-target effects associated with CRISPR/Cas9 genome editing. It is concluded that CAR-T cells modified by CRISPR/Cas9 significantly improved antitumor efficacy in a cost-effective manner that provides opportunities for novel cancer immunotherapies.
A series of novel dimethoxyindanone embedded spiropyrrolidines were synthesized in ionic liquid, [bmim]Br and were evaluated for their inhibitory activities towards cholinesterases. Among the spiropyrrolidines, compound 4f exhibited the most potent activity with an IC50 value of 1.57 µM against acethylcholinesterase (AChE). Molecular docking simulation for the most active compound was employed with the aim of disclosing its binding mechanism to the active site of AChE receptor.
Ecosystem functioning is dependent a lot on large mammals, which are, however, vulnerable and facing extinction risks due to human impacts mainly. Megafauna of Asia has been declining for a long, not only in numbers but also in their distribution ranges. In the current study, we collected information on past and current occurrence and distribution records of Asia's megafauna species. We reconstructed the historical distribution ranges of the six herbivores and four carnivores for comparison with their present ranges, to quantify spatially explicit levels of mega-defaunation. Results revealed that historically the selected megafauna species were more widely distributed than at current. Severe range contraction was observed for the Asiatic lion, three rhino species, Asian elephant, tigers, and tapirs. Defaunation maps generated have revealed the vanishing of megafauna from parts of the East, Southeast, and Southwest Asia, even some protected Areas losing up to eight out of ten megafaunal species. These defaunation maps can help develop future conservation policies, to save the remaining distribution ranges of large mammals.
Wheat (Triticum aestivum L.) production is significantly altered by the infestation of sucking insects, particularly aphids. Chemical sprays are not recommended for the management of aphids as wheat grains are consumed soon after crop harvests. Therefore, determining the susceptibility of different wheat genotypes and selecting the most tolerant genotype could significantly lower aphid infestation. This study evaluated the susceptibility of six different wheat genotypes ('Sehar-2006', 'Shafaq-2006', 'Faisalabad-2008', 'Lasani-2008', 'Millat-2011' and 'Punjab-2011') to three aphid species (Rhopalosiphum padi Linnaeus, Schizaphis graminum Rondani, Sitobion avenae Fabricius) at various growth stages. Seed dressing with insecticides and plant extracts were also evaluated for their efficacy to reduce the incidence of these aphid species. Afterwards, an economic analysis was performed to compute cost-benefit ratio and assess the economic feasibility for the use of insecticides and plant extracts. Aphids' infestation was recorded from the seedling stage and their population gradually increased as growth progressed towards tillering, stem elongation, heading, dough and ripening stages. The most susceptible growth stage was heading with 21.89 aphids/tiller followed by stem elongation (14.89 aphids/tiller) and dough stage (13.56 aphids/tiller). The genotype 'Punjab-2011' recorded the lower aphid infestation than 'Faisalabad-2008', 'Sehar-2006', 'Lasani-2008' and 'Shafaq-2006'. Rhopalosiphum padi appeared during mid-February, whereas S. graminum and S. avenae appeared during first week of March. Significant differences were recorded for losses in number of grains/spike and 1000-grain weight among tested wheat genotypes. The aphid population had non-significant correlation with yield-related traits. Hicap proved the most effective for the management of aphid species followed by Hombre and Husk among tested seed dressers, while Citrullus colocynthis L. and Moringa oleifera Lam. plant extracts exhibited the highest efficacy among different plant extracts used in the study. Economic analysis depicted that use of Hombre and Hicap resulted in the highest income and benefit cost ratio. Therefore, use of genotype Punjab-2011' and seed dressing with Hombre and Hicap can be successfully used to lower aphid infestation and get higher economic returns for wheat crop.
Pure and manganese-doped titanium dioxide nanoparticles (MnTiO2-NPs) were synthesized by the defect-oriented hydrothermal approach. The synthesized material was then characterized by X-ray diffraction (XRD), Scanning electron microscopy (SEM), Energy dispersive X-ray spectroscopy (EDX), and UV-visible spectroscopy (UV-Vis). The agar well diffusion method assessed the antibacterial efficiency of TiO2 and MnTiO2-NPs against E. coli and S. aureus. Zone of inhibition (ZOI) formed by pure TiO2 was observed as 12 mm and 11.5 mm against E. coli and S. aureus, while for MnTiO2-NPs it was observed as 19 mm (E. coli) and 21 mm (S. aureus). The concentration of synthesized nanoparticles (10 mg/ml, and 20 mg/ml) was used for antibacterial studies. The efficacy of the pure and MnTiO2-NPs as an active photocatalyst for the degradation of methylene blue (MB) dye was also assessed using a UV light. It was observed that the photodegradation efficiency of 1 g of MnTiO2-NPs was higher than the same amount of pure TiO2. The results suggest that the photocatalyst concentration directly impacts the photodegradation of MB dye. The pH value was found to influence the photodegradation of MB dye at higher pH values. Based on the obtained results, MnTiO2-NPs were observed as a promising agent for microbial resistance and water remediation.
Molecular hydrogen is renowned as an odorless and colorless gas. The recommendations developed by China suggest that the inhalation of hydrogen molecules is currently advised in COVID-19 pneumonia treatment. The therapeutic effects of molecular hydrogens have been confirmed after numerous clinical trials and animal-model-based experiments, which have expounded that the low molecular weight of hydrogen enables it to easily diffuse and permeate through the cell membranes to produce a variety of biological impacts. A wide range of both chronic and acute inflammatory diseases, which may include sepsis, pancreatitis, respiratory disorders, autoimmune diseases, ischemia-reperfusion damages, etc. may be treated and prevented by using it. H2 can primarily be inoculated through inhalation, by drinking water (which already contains H2), or by administrating the injection of saline H2 in the body. It may play a pivotal role as an antioxidant, in regulating the immune system, in anti-inflammatory activities (mitochondrial energy metabolism), and cell death (apoptosis, pyroptosis, and autophagy) by reducing the formation of excessive reactive O2 species and modifying the transcription factors in the nuclei of the cells. However, the fundamental process of molecular hydrogen is still not entirely understood. Molecular hydrogen H2 has a promising future in therapeutics based on its safety and possible usefulness. The current review emphasizes the antioxidative, anti-apoptotic, and anti-inflammatory effects of hydrogen molecules along with the underlying principle and fundamental mechanism involved, with a prime focus on the coronavirus disease of 2019 (COVID-19). This review will also provide strategies and recommendations for the therapeutic and medicinal applications of the hydrogen molecule.