Aspergillus flavus is a fungal pathogen which infects maize crops and produces aflatoxin thus bringing about huge losses in crop production. Developing biocontrol agents against Aspergillus flavus has been the best strategy for the control of contamination in the fields. The aim of this study was to evaluate the biocontrol potential of Aspergillus giganteus against A. flavus by in vitro coculture studies. The effect of antagonism was studied by varying the carbon and nitrogen sources and under different interacting conditions of pH, temperature and water activities. The conidia production by A. flavus during coculture conditions favourable for antagonism was also assessed. A significantly notable growth inhibition of about 86.1% was brought about by A. giganteus in the coculture, which surrounded the mycelia of pathogenic A. flavus, arresting its growth. A maximum inhibition of 86.1% was observed when sucrose was used as the carbon source and a significantly higher inhibition of 90.93% was seen when beef extract was used as the nitrogen source. Among the different temperatures tested, the highest inhibition was observed at 30°C which was 87.43%. An increasing trend in the inhibitions were seen with decrease in pH and water activity (a w), where, the highest inhibition was 89.75% for pH 6.0 and 94.03% for a w 0.846. Drastic reductions in conidial number and halting of sclerotia production was observed in coculture clearly suggesting that A. giganteus will serve to be a potent and promising biocontrol strain under different environmental conditions.
Cellular responses to inputs that vary both temporally and spatially are determined by complex relationships between the components of cell signaling networks. Analysis of these relationships requires access to a wide range of experimental reagents and techniques, including the ability to express the protein components of the model cells in a variety of contexts. As part of the Alliance for Cellular Signaling, we developed a robust method for cloning large numbers of signaling ORFs into Gatewayா entry vectors, and we created a wide range of compatible expression platforms for proteomics applications. To date, we have generated over 3000 plasmids that are available to the scientific community via the American Type Culture Collection. We have established a website at www.signaling-gateway.org/data/plasmid/ that allows users to browse, search, and blast Alliance for Cellular Signaling plasmids. The collection primarily contains murine signaling ORFs with an emphasis on kinases and G protein signaling genes. Here we describe the cloning, databasing, and application of this proteomics resource for large scale subcellular localization screens in mammalian cell lines. Molecular & Cellular Proteomics 6: 413-424, 2007.Analysis of cross-talk between signaling pathways when mammalian cells are challenged with multiple ligand stimuli and the development of molecular models that describe signal integration and processing provide key insight to cellular signaling mechanisms and the regulation of cellular function (1, 2). Two strategic requirements in projects of this nature are the identification of the protein components in a given model system (the so-called "parts list") and the ability to modulate the expression and function of these protein components. At the outset of the Alliance for Cellular Signaling (AfCS) 1 project (1), reagent availability was a central issue, and in the case of cDNA clones, there was no publicly accessible repository of validated mouse sequences. Moreover the nature of the project demanded the establishment of standardized methodology for the isolation of cDNAs and their expression in a variety of contexts. The emergence of recombination-based cloning technologies was timely and allowed the development of a robust cloning platform. We adopted Invitrogen's Gatewayா system that allows facile transfer of DNA segments into multiple expression platforms while maintaining orientation and reading frame register (3-5).Several reports have described efforts to generate genomewide collections of cDNAs (6, 7) and the production of "ORFeome" sets of sequence-validated open reading frames (8 -10). The generation of "ORF-only" clones in these latter efforts is key for downstream proteomics applications that require expression of proteins with N-or C-terminal fusion tags (11)(12)(13)(14). The AfCS cloning effort has focused on genes involved in cell signaling and therefore does not approach the scale of these genome-wide projects. However, as the described ORFeome cloning projects have concentrated on model org...
Fungal infections are more predominant in agricultural and clinical fields. Aspergillosis caused by Aspergillus fumigatus leads to respiratory failure in patients along with various illnesses. Due to the limitation of antifungal therapy and antifungal drugs, there is an emergence to develop efficient antifungal compounds from natural sources to cure and prevent fungal infections. The present study deals with the investigation of the mechanism of active compounds from our candidate agonist Aspergillus giganteus for aspergillosis. The integrity of treated Aspergillus fumigatus cell membrane and nuclear membrane was analyzed by determining the release of cellular materials. The antagonistic potential of antifungal compounds on the pathogen was confirmed by SEM analysis. The effective concentration of antifungal compounds (AFCs) was found to be 250µg/ml. The GC-MS profiling has revealed the bioactive metabolites responsible for the antagonistic nature of Aspergillus giganteus. The bioavailability and toxicological properties of pathogenesis related proteins have proved the efficiency of pharmacokinetic properties of selected compounds. Interaction of sarcin, thionin, chitinase and its derivatives from Aspergillus giganteus with the virulence proteins of UDP-N-acetylglucosamine pyrophosphorylase, N-myristoyl transferase and Chitinase have proved the druggable nature of the antifungal compounds.
Several medicinal plants have the potential to be a promising alternative pharmacological therapy for a variety of human illnesses. Many insects, including mosquitoes, are important vectors of deadly pathogens and parasites, which in the world’s growing human and animal populations can cause serious epidemics and pandemics. Medicinal plants continue to provide a large library of phytochemicals, which can be used to replace chemically synthesized insecticides, and utilization of herbal product-based insecticides is one of the best and safest alternatives for mosquito control. Identifying new effective phyto-derived insecticides is important to counter increasing insect resistance to synthetic compounds and provide a safer environment. Solanum genus (Solanaceae family or nightshades) comprises more than 2500 species, which are widely used as food and traditional medicine. All research publications on insecticidal properties of Solanaceae plants and their phytoconstituents against mosquitoes and other insects published up to July 2020 were systematically analyzed through PubMed/MEDLINE, Scopus, EBSCO, Europe PMC, and Google Scholar databases, with focus on species containing active phytoconstituents that are biodegradable and environmentally safe. The current state of knowledge on larvicidal plants of Solanum species, type of extracts, target insect species, type of effects, name of inhibiting bioactive compounds, and their lethal doses (LC50 and LC90) were reviewed in this study. These studies provide valuable information about the activity of various species of Solanum and their phytochemical diversity, as well as a roadmap for optimizing select compounds for botanical repellents against a variety of vectors that cause debilitating and life-threatening human diseases.
Occurrence and intensity of systemic invasive fungal infections have significantly risen in recent decades with large amount of mortality and morbidity rates at global level. Treatment therapy lies on the current antifungal interventions and are often limited due to the emergence of resistance to antifungal agents. Chemosensitization of fungal strains to the conventional antimycotic drugs are of growing concern. Current antifungal drugs often have been reported with poor activity and side effects to the host and have a few number of targets to manifest their efficacy on the pathogens. Indiscriminately, the aforementioned issues have been easily resolved by the development of new intervention strategies. One such approach is to employ combinational therapy that has exhibited a great level of inhibitions than that of a single compound. Chemosensitization of pathogenic mycoses to commercial antifungal drugs could be drastically enhanced by co-application of chemosensitizers along with the conventional drugs. Chemosensitizers could address the resistance mechanisms evolved in the pathogenic fungi and targeting the system to make the organism susceptible to commercially and clinically proven antifungal drugs. However, this strategy has not been overreached to the greater level, but it needs much attention to fight against not only with the pathogen but combat the resistance mechanisms of pathogens to drugs. Natural compounds including plant compounds and microbial proteins act as potential chemosensitizers to break the resistance in mycoses. Aspergillus giganteus, a filamentous fungus, is known to produce a cysteine rich extracellular protein called as antifungal protein (AFP). AFP has shown enhanced efficacy against several filamentous and non-filamentous fungal pathogens. On the basis of the reported studies on its targeted potential against pathogenic mycoses, AFP would be fabricated as a good chemosensitizer to augment the fungicidal efficacy of commercial antimycotic drugs. This paper reviews on breakthrough in the discovery of antifungal drugs along with the resistance patterns of mycoses to commercial drugs followed by the current intervention strategies applied to augment the fungicidal potential of drugs.
Aspergillosis is a dreadful fungal infection and are more predominant in clinical fields. Due to the limitation of antifungal drugs, there is an emergence to develop efficient antifungal compounds from natural sources. Hence, the present study deals with the validation of active compounds from Aspergillus giganteus against aspergillosis causing Aspergillus fumigatus. The most prominent antifungal proteins in Aspergillus giganteus are sarcin, thionin and chitinase. Initially, the bioavailability and toxicological properties of sarcin, thionin, chitinase and their derivatives were screened. The molecular interaction of the screened antifungal proteins against the target proteins (UDP-N-acetylglucosamine pyrophosphorylase, N-myristoyl transferase and Chitinase) of Aspergillus fumigatus was performed using Schrodinger module. The antagonistic potential of antifungal compounds on the pathogen was confirmed by SEM. The integrity of Aspergillus fumigatus cell membrane and nuclear membrane treated with antifungal compounds were analysed by determining the release of cellular materials. Further, the GC-MS profiling of volatile bioactive compounds were analysed. The results have proved the efficiency of selected compounds for their pharmacokinetic properties. Molecular interactions of selected compounds from Aspergillus giganteus with the virulence proteins of Aspergillus fumigatus have exhibited a good glide score and their druggable nature. The SEM analysis have envisaged the shrunken and damaged spores of A. fumigatus treated with antifungal compounds. The effective concentration of antifungal compounds (AFCs) was found to be 250 µg/ml (p<0.0001). The GC-MS profiling has revealed the volatile bioactive metabolites present in Aspergillus giganteus. Conclusively, the selected antagonists from Aspergillus giganteus can be a good drug candidate to treat aspergillosis.
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