Aspergillus Fumigatus Extract Research Articles

Larvicidal Effect of Spores and Metabolites Extracts of Aspergillus Fumigatus against Culex Mosquito Larvae

Larvicidal Effect of Spores and Metabolites Extracts of Aspergillus Fumigatus against Culex Mosquito Larvae

Dual inhibition of airway inflammation and fibrosis by common β cytokine receptor blockade

BackgroundSevere asthmatics can present with eosinophilic type 2 (T2), neutrophilic or mixed inflammation that drives airway remodeling and exacerbations and represents a major treatment challenge. The common beta (βc) receptor signals for three cytokines GM-CSF, IL-5 and IL-3, that collectively mediate T2 and neutrophilic inflammation. ObjectivesTo determine the pathogenesis of βc receptor mediated inflammation and remodeling in severe asthma, and to investigate βc antagonism as a therapeutic strategy for mixed granulocytic airway disease. Methodsβc gene expression was analysed in bronchial biopsies from mild/moderate and severe asthmatics. House dust mite extract (HDM) and Aspergillus fumigatus extract (ASP) models were used to establish asthma-like pathology and airway remodeling in human βc transgenic mice. Lung tissue gene expression was analysed by RNA-sequencing. The monoclonal antibody CSL311 targeting the shared cytokine binding site of βc was used to block βc signaling. Resultsβc gene expression was increased in severe asthmatics. CSL311 potently reduced lung neutrophils, eosinophils and interstitial macrophages, and improved airway pathology and lung function in the acute steroid resistant HDM model. Chronic intranasal ASP exposure induced airway inflammation and fibrosis and impaired lung function that was inhibited by CSL311. CSL311 normalised the ASP-induced fibrosis-associated extracellular matrix gene expression network and strongly reduced signatures of cellular inflammation in the lung. Conclusionsβc cytokines drive steroid-resistant mixed myeloid cell airway inflammation and fibrosis. The anti-βc antibody CSL311 effectively inhibits mixed T2/neutrophilic inflammation and severe asthma-like pathology and reverses fibrosis gene signatures induced by exposure to commonly encountered environmental allergens.

Identification and production of novel potential pathogen-specific biomarkers for diagnosis of histoplasmosis.

Immunological assays based on the detection of circulating fungal biomarkers are helpful in clinical practice for diagnosing invasive fungal infections. Some of these targeting antigenic components are common to several different fungi. Histoplasmosis is a mycosis caused by the dimorphic fungus Histoplasma capsulatum, which in recent years has gained significant relevance due to the increase in the population susceptible to developing severe clinical forms, including those living with HIV/AIDS. Immunological tests that detect cell wall polysaccharide antigens are among the most used laboratory techniques for diagnosing this mycosis. However, none have shown adequate performance, and cross-reactivity with other fungal pathogens may be observed. Considering that there is a real need to improve the sensitivity and specificity of current diagnostic methods, we explored a novel strategy for the identification of H. capsulatum-specific antigens (Hc_Ags) that could be detected in clinical samples during infection based on a computational analysis model that included the application of bioinformatics tools and the analysis of experimental data from transcriptomics and proteomics. The Hc_Ags identified were expressed and purified by eukaryotic and prokaryotic systems. First, the Hc_Ags were used in an in-house immunization model in mice (BALB/c) to obtain Hc_Ag-specific polyclonal antibodies (Hc_Ag_PAb). Then, the presence of these antigens in H. capsulatum-yeast culture extracts and the specificity of Hc_Ag_PAb against culture extracts of Candida albicans, Aspergillus fumigatus, Cryptococcus neoformans, Fusarium spp., and Paracoccidioides brasiliensis were confirmed. Finally, we demonstrated the immunoreactivity of these Hc_Ag-specific polyclonal antibodies with urine samples from patients previously diagnosed with histoplasmosis.IMPORTANCEHistoplasmosis is considered one of the most important mycoses due to the increasing number of individuals susceptible to develop severe clinical forms, particularly those with HIV/AIDS or receiving immunosuppressive biological therapies, the high mortality rates reported when antifungal treatment is not initiated in a timely manner, and the limitations of conventional diagnostic methods. In this context, there is a clear need to improve the capacity of diagnostic tools to specifically detect the fungal pathogen, regardless of the patient's clinical condition or the presence of other co-infections. The proposed novel pathogen-specific biomarkers have the potential to be used in immunodiagnostic platforms and antifungal treatment monitoring in histoplasmosis. In addition, the bioinformatics strategy used in this study could be applied to identify potential diagnostic biomarkers in other models of fungal infection of public health importance.

Abnormal adenosine metabolism of neutrophils inhibits airway inflammation and remodeling in asthma model induced by Aspergillus fumigatus

BackgroundNeutrophils consume a large amount of energy when performing their functions. Compared with other white blood cells, neutrophils contain few mitochondria and mainly rely on glycolysis and gluconeogenesis to produce ATP. The inflammatory site is hypoxic and nutrient poor. Our aim is to study the role of abnormal adenosine metabolism of neutrophils in the asthmatic airway inflammation microenvironment.MethodIn this study, an asthma model was established by intratracheal instillation of Aspergillus fumigatus extract in Ecto-5'-Nucleotidase (CD73) gene–knockout and wild-type mice. Multiple analyses from bronchoalveolar lavage fluid (BALF) were used to determine the levels of cytokines and chemokines. Immunohistochemistry was used to detect subcutaneous fibrosis and inflammatory cell infiltration. Finally, adenosine 5’-(α, β-methylene) diphosphate (APCP), a CD73 inhibitor, was pumped subcutaneously before Aspergillus attack to observe the infiltration of inflammatory cells and subcutaneous fibrosis to clarify its therapeutic effect.ResultPAS staining showed that CD73 knockout inhibited pulmonary epithelial cell proliferation and bronchial fibrosis induced by Aspergillus extract. The genetic knockdownof CD73 significantly reduced the production of Th2 cytokines, interleukin (IL)-4, IL-6, IL-13, chemokine (C–C motif) ligand 5 (CCL5), eosinophil chemokine, neutrophil IL-17, and granulocyte colony-stimulating factor (G-CSF). In addition, exogenous adenosine supplementation increased airway inflammation. Finally, the CD73 inhibitor APCP was administered to reduce inflammation and subcutaneous fibrosis.ConclusionElevated adenosine metabolism plays an inflammatory role in asthma, and CD73 could be a potential therapeutic target for asthma.

Efficacy of Silver Nanoparticle-Coated Crude Extract of Aspergillus fumigatus Against Bacterial Infections from Surgical Sites Wound

Efficacy of Silver Nanoparticle-Coated Crude Extract of Aspergillus fumigatus Against Bacterial Infections from Surgical Sites Wound

Exploration of the chemical constituents and its antioxidant, antibacterial activities of endophytic fungi isolated from the medicinal plant Dillenia indica.

Assam, India being the pool for ethnomedicinal plants harbors diverse endophytic fungi constituting major bioactive metabolites. The present study was designed to screen the antioxidant, antibacterial activities along with the chemical constituents of the endophytic fungi isolated from the fruits of Dillenia indica (commonly known as Otenga in Assam). Screening of such metabolic compounds and their antioxidant, antibacterial activities can have tremendous potential in suppressing certain diseases. Agar well diffusion method has been used to carry out the antibacterial assay against three pathogenic bacteria two gram positive [Bacillus subtilis (MTCC No. 441); Staphylococcus aureus (MTCC No. 740)] and one gram negative [Escherichia coli (MTCC No. 739)]. Aspergillus fumigatus of ethyl acetate extract showed a prominent activity against Staphylococcus aureus followed by Aspergillus flavus and Aspergillus niger. Antioxidants have the potential to neutralize and inhibit the action of free radicals. The highest scavenging activity was exhibited by ethyl acetate extract of Aspergillus fumigatus in DPPH assay. Furthermore, the phytochemical screening revealed the presence of flavonoids, alkaloids, terpenoids and saponins. Result showed that ethyl acetate extract of Aspergillus fumigatus showed the highest phenolic content (236.81 ± 0.2mg.g-1) and least was shown by Aspergillus flavus (92.12 ± 1.4mg.g-1). Total flavonoids content for Aspergillus fumigatus (39.08 ± 0.2mg.g-1) was found to be highest compared to other isolates. Molecular identification of the endophytic fungus showing highest activity was done based on 18S rRNA. The sequenced was submitted in Genbank with accession number MH540721 showing high similarities with Aspergillus fumigatus strain 3,162,954. A. fumigatus strain is subjected to GC/MS analysis that revealed the chemical constituents 2-isopropyl-5-methyl-1-heptanol, dodecane, 1-fluoro-pentanoic acid, 2-ethylhexyl ester, 1-octanol, 2-butyl-1-dodecanol. Thus, the present work reveals that endophytic fungi colonizing in ethnomedicinal plant Dillenia indica could be a promising source for antioxidant and antibacterial activity. Further work is needed to add value in various therapeutic and pharmaceutical fields.

Bimetallic Assembled Silver Nanoparticles Impregnated in Aspergillus fumigatus Extract Damage the Bacterial Membrane Surface and Release Cellular Contents

The bactericidal effects of nanomaterials play an essential role in cytoplasmic leakage, leading to bacterial cell death. In this study, silver nanoparticles (AgNPs) were synthesized using a fungal extract of Aspergillus fumigatus (A. fumigatus). The physicochemical properties of the bare and myco-synthesized AgNPs (MS-AgNPs) were examined by X-ray diffraction (XRD), Fourier transform infrared (FT-IR) spectroscopy, ultraviolet–visible (UV–vis) spectroscopy, scanning electron microscopy (SEM), energy dispersive X-ray spectroscopy (EDX), and transmission electron microscopy (TEM). XRD revealed the crystalline structure of the prepared NPs. The FTIR spectrum of the MS-AgNPs revealed the presence of the stretching vibrations of hydroxyl (−OH) and carbonyl groups (C=O). The UV results showed absorption from 450 nm to 590 nm, confirming the synthesis of the AgNPs. SEM and TEM showed rough cubic shapes (spheres), 20–60 nm in size, while EDX confirmed the presence of 60% Ag in the sample. The MS-AgNPs revealed the highest antibacterial activity against Staphylococcus aureus, with a zone of inhibition of 18.21 ± 2.1 mm, followed by Shigella dysenteriae and Salmonella typhi. The bimetallic-AgNPs played a vital role in cell membrane damage and the release of cellular contents, specifically nucleic acids and proteins. These results suggest that MS-AgNPs have promising antimicrobial capabilities and might be beneficial for an extensive array of biological applications.

Basophil Activation Test With Aspergillus Molecules: The Case for ABPA.

BackgroundAllergic bronchopulmonary aspergillosis (ABPA) is an underestimated allergic disease due to Aspergillus fumigatus (AF). The main diagnostic criteria for ABPA rely on the evaluation of immunoglobulin (Ig) E and IgG responses to AF extracts, although these cannot discriminate AF-sensitization from ABPA.ObjectivesTo evaluate the performance of cellular functional assays with extract and molecular AF allergens in ABPA.MethodsA prospective cohort of 67 patients (6 ABPA) was investigated with basophil activation test (BAT) with AF extract. Twelve patients were further investigated for BAT responses to molecular AF components: Asp f 1, Asp f 2, Asp f 3, Asp f 4, and Asp f 6.ResultsBAT with AF extract with an optimized cutoff displayed 100% sensitivity and 77.6% specificity for ABPA diagnosis. Among patients with positive BAT to AF, BAT with Asp f 4 was significantly higher in ABPA patients at 10 ng/mL (mean basophil stimulation index 10.56 in ABPA vs. 1.24 in non-ABPA patients, p = 0.0002).ConclusionBAT with AF is a promising diagnostic biomarker in the context of suspected ABPA, which can be further improved with AF molecular allergens, especially Asp f 4.

Biosynthesis of Antimicrobial Silver Nanoparticles against Gram-Negative and Gram-positive Bacteria by the Entophytic Fungus Aspergillus Fumigatus

Multi drug resistance is increasing day by day due to misuse of antibiotics. Several potent metabolites are produced by fungi. Synthesis of silver nanoparticle (AgNPs) Due to its simple, harmless, time-saving, and cost-effective characteristics, it has acquired great popularity in recent years. A variety of analytical techniques were used to synthesize AgNPs from Aspergillus fumigatus extracts, including X-ray Diffraction (XRD), Fourier Transform Infrared Spectroscopy (FTIR), Scanning Electron Microscopy (SEM), and Transmission Electron Microscopy (TEM). The effect of synthesis AgNPs and crude extract noted against different bacterial pathogens. Maximum antibacterial activity were noticed against tested bacteria by both fungal crude extract and (AgNPs) Maximum antibacterial activity of Ethyl acetate crude extract at 50µl concentration (12mg/1ml DMSO) showed (15mm) zone of inhibition against E.coli. While minimum antibacterial activity of Ethyl crude extract at 50µl concentration was observed against S.typhi (12mm). Highest antibacterial activity of Ethyl acetate crude extract at 100µl was noted against E.coli which showed (20mm) zone of inhibition. While (17mm) zone of inhibition was observed against S.typhi at 100µl concentration Ethyl acetate crude extract and AgNPs (25mm) zone of inhibition was observed against E.coli at 100µl concentration Ethyl acetate crude extract and AgNPs respectively. During UV-visible spectroscopy, surface Plasmon Resonance (SPR) was observed at 432 nm, which confirmed the synthesis of AgNPs. The SEM micrograph demonstrated the spherical shape of AgNPs. The results of FTIR research revealed that phenolic, carboxyl, and hydroxyl groups played a crucial role in the reduction of Ag+ ions into AgNPs, while amide linkage and amino acids stabilized AgNPs. AgNPs synthesized were XRD peaks that revealed phase purity, size, internal crystalline structure, and nature. In the pharmaceutical and medical fields, AgNPs synthesized from Aspergillus fumigatus extract could be of great importance. While, the combination of AgNPs and crude extract Aspergillus fumigatus enhances their antimicrobial effect which increase their importance in future studies Keywords: Fungal secondary metabolites, silver nanoparticles, crude extract, MDR pathogen bacteria , SEM, TEM, UV, FTIR, XRD.

Identification of Antibacterial Metabolites from Endophytic Fungus Aspergillus fumigatus, Isolated from Albizia lucidior Leaves (Fabaceae), Utilizing Metabolomic and Molecular Docking Techniques

The rapid spread of bacterial infection caused by Staphylococcus aureus has become a problem to public health despite the presence of past trials devoted to controlling the infection. Thus, the current study aimed to explore the chemical composition of the extract of endophytic fungus Aspergillus fumigatus, isolated from Albizia lucidior leaves, and investigate the antimicrobial activity of isolated metabolites and their probable mode of actions. The chemical investigation of the fungal extract via UPLC/MS/MS led to the identification of at least forty-two metabolites, as well as the isolation and complete characterization of eight reported metabolites. The antibacterial activities of isolated metabolites were assessed against S. aureus using agar disc diffusion and microplate dilution methods. Compounds ergosterol, helvolic acid and monomethyl sulochrin-4-sulphate showed minimal inhibitory concentration (MIC) values of 15.63, 1.95 and 3.90 µg/mL, respectively, compared to ciprofloxacin. We also report the inhibitory activity of the fungal extract on DNA gyrase and topoisomerase IV, which led us to perform molecular docking using the three most active compounds isolated from the extract against both enzymes. These active compounds had the required structural features for S. aureus DNA gyrase and topoisomerase IV inhibition, evidenced via molecular docking.

Resveratrol attenuates inflammation and apoptosis through alleviating endoplasmic reticulum stress via Akt/mTOR pathway in fungus-induced allergic airways inflammation

BackgroundResveratrol has shown pleiotropic effects against inflammation and oxidative response. The present study aimed to investigate the effects and mechanisms of resveratrol on fungus-induced allergic airway inflammation. MethodsFemale BALB/c mice were injected intraperitoneally with Aspergillus fumigatus (Af) extract emulsified with aluminum on day 0 and 7 and intranasally challenged with Af extracts on day 14 and 15. Resveratrol or dexamethasone or a vehicle was injected intraperitoneally 1 h before each challenge. Mice were sacrificed for serum, bronchoalveolar lavage fluid (BALF), and lungs 24 h after the last challenge. The control group was administered with saline. BEAS-2B was used for the experiments in vitro that Af-exposed airway epithelial cells. ResultsResveratrol and dexamethasone attenuated the airway inflammation and eosinophilia, and reduced not only the production of IL-4, IL-5, and IL-13 in the BALF and lung tissues but also the mRNA levels of lung IL-6, TNF-α, and TGF-β induced by Af challenge (P < 0.05). Furthermore, Af-induced lung endoplasmic reticulum (ER) stress-related proteins PERK, CHOP, and GRP78 and the apoptosis markers including cleaved caspase-3 and cleaved caspase-7 were both suppressed significantly by resveratrol (P < 0.05). In vitro, activation of ER stress and the Akt/mTOR pathway in Af-exposed BEAS-2B cells were effectively ameliorated by resveratrol. Inhibition of the Akt/mTOR pathway using LY294002 suppressed the ER stress while ER stress inhibitor 4-PBA decreased the apoptosis in Af-exposed BEAS-2B cells. ConclusionsOur findings collectively revealed that resveratrol alleviated the Af-exposed allergic inflammation and apoptosis through inhibiting ER stress via Akt/mTOR pathway, exerting therapeutic effects on the fungus-induced allergic lung disorder.

Fibrin(ogen) Mechanistically Contributes to Atopic Dermatitis Pathogenesis and Allergic Sensitization

Adults with atopic dermatitis (AD) have an increased risk for thromboembolic events. While coagulation is critical for almost all healing responses, dysregulated clotting and fibrin(ogen) deposition lead to inflammation that exacerbates tissue damage and impairs tissue repair. While plasma fibrin(ogen) has been associated with established asthma, fibrin(ogen)'s mechanistic role in AD pathogenesis and allergic sensitization has not been investigated. Here we show that fibrinogen plays a critical role in a murine model atopic dermatitis pathogenesis by markedly attenuating disease severity, barrier dysfunction, and allergic sensitization.To determine if fibrinogen impacts AD pathogenesis, we used mice with complete fibrinogen deficiency (Fib -/-), partial deficiency (Fib +/-), and wildtype controls (Fib WT) in our established model of AD. This model uses repeated cutaneous allergen sensitization with heat-killed Aspergillus fumigatus (Asp) extract. Notably, Fib +/- mice produce approximately half as much fibrinogen as Fib WT mice. After each patch and at sacrifice each mouse underwent an objective disease severity assessment as well as transepidermal water loss (TEWL) measurements to assess skin barrier function. Our results show that complete and partial fibrinogen deficiency abrogated AD development. We found that Fib -/- and Fib +/- mice had markedly attenuated TEWL (P-value <0.0001) and disease severity (P-value <0.0001) compared with controls (Figure 1). In addition, we found that the TEWL and disease severity in Fib -/- and Fib +/- were comparable to unchallenged controls. There was also no difference in either outcome between Fib -/- and Fib +/- mice within the experimental group. Lastly, none of the Fib -/- and Fib +/- mice developed spontaneous bleeding.Next, to elucidate the impact of fibrinogen on allergic sensitization we measured plasma allergen specific immunoglobulin E (sIgE) to Asp in all mice in our AD model at sacrifice. We found that Fib -/- and Fib +/- mice had significantly lower Asp sIgE compared with controls (p-value <0.0001 and 0.0002, respectively; Figure 2). In addition, we found that the amount of sIgE to Asp produced was comparable between Fib -/- and Fib +/- mice as well as with unchallenged controls.Thus, our findings suggest that fibrin(ogen) plays a critical mechanistic role in driving AD development and allergic sensitization. While fibrinogen's role in established allergic disease has been shown, these novel findings suggest a critical role for fibrinogen in allergic disease pathogenesis which may allow for additional therapeutic targets to treat atopic dermatitis and other allergic diseases. [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

TSLP rather than IL-33 signaling drives intestinal mucosal mast accumulation and allergen-induced diarrhea in mice following epicutaneous sensitization to food allergen.

Abstract Background: A major route of sensitization to food allergen is through an impaired skin barrier. IL33 and TSLP have both been implicated in epicutaneous sensitization and development of food allergy. Method: We assessed the respective contributions of TSLP and IL33 to the development of food allergy in TSLP and IL33 receptor deficient mice following epicutaneous food allergen sensitization, where mice were exposed to thrice weekly skin patches of either saline, OVA or a combination of OVA and Aspergillus fumigatus extract (ASP). Results: ASP+OVA-patched but not OVA-patched mice developed an atopic dermatitis (AD)-like skin phenotype, while sensitization to OVA occurred in both OVA and OVA+ASP patched mice. OVA-specific IgE levels were significantly lower in OVA(±ASP)-patched TSLPR−/− mice compared to wild type mice and ST2−/− mice. Repeated intragastric challenges with 50mg of OVA, induced intestinal accumulation of mast cells but not ILC2s in OVA(±ASP)-patched WT and ST2−/− mice, and to a much lesser degree in TSLPR−/− mice. While OVA-induced mast cell degranulation assessed by measuring MCPT1 blood levels and the development of food allergy (diarrhea occurrences) was observed in almost all WT mice and two thirds of OVA+ASP patched ST2−/− mice, all TSLPR−/− mice were protected from developing food allergy and had significantly lower MCPT1 levels. Conclusion: Epicutaneous sensitization to food allergen and subsequent development of food allergy does not require atopic dermatitis skin lesions and is dependent on TSLP, suggesting that prophylactic targeting of TSLP may be useful in mitigating the development of food allergy early in life.

Aspergillus fumigatus-derived demethoxyfumitremorgin C inhibits proliferation of PC3 human prostate cancer cells through p53/p21-dependent G1 arrest and apoptosis induction

Human prostate cancer is the second most frequently diagnosed cancer worldwide, and its incidence rate continues to increase. Advanced prostate cancer is more difficult to treat than early forms due to its chemotherapy resistance. There is need for more effective agents that can inhibit the progression of advanced prostate cancer. Demethoxyfumitremorgin C (DMFTC) was isolated from the fermentation extract of the marine fungus Aspergillus fumigatus. Antiproliferative activity of DMFTC against human prostate cancer PC3 cells was examined through cell cycle analysis by flow cytometry, the fluorescent nuclear imaging analysis with propidium iodide (PI), and proteins expression related to cell cycle arrest and apoptosis were investigated via Western blotting. DMFTC inhibited PC3 cells growth through G1 phase cell cycle arrest and apoptosis induction. It activated the tumor suppressor p53 and the Cdk inhibitor p21, which regulate the cell progression into the G1 phase. Additionally, PI-positive late apoptotic non-viable cells were increased and the expression levels of the G1-positive downstream regulators cyclin D, cyclin E, Cdk2, and Cdk4 were decreased by DMFTC treatment. These results suggest that DMFTC induces G1 arrest and apoptosis induction through regulation of p53/p21-dependent cyclin-Cdk complexes, and it may be a useful therapeutic agent for the treatment of human advanced prostate cancer.

Fungal extracts stimulate solitary chemosensory cell expansion in noninvasive fungal rhinosinusitis.

Solitary chemosensory cells (SCCs) are rare epithelial cells enriched in nasal polyps and are the primary source of interleukin-25 (IL-25), an innate cytokine eliciting T-helper 2 (Th2) immune response. Although it is proposed that SCCs are stimulated by antigens released by upper airway pathogens, the exogenous triggers of human SCCs remain elusive. We studied patients with noninvasive fungal rhinosinusitis to determine whether extracts of Aspergillus fumigatus and Alternaria alternata stimulate SCC proliferation as an early event in type 2 inflammation. Multicolor flow cytometry, immunofluorescence, and enzyme-linked immunoassay were used to interrogate mucosa from patients with mycetomas and allergic fungal rhinosinusitis (AFRS) for SCCs and IL-25. Primary sinonasal epithelial cells from AFRS patients and noninflamed inferior turbinates were stimulated with fungal extracts for 72 hours, and SCC population frequency as well as mitotic activity were quantified using flow cytometry. SCCs producing IL-25 are enriched in inflamed mucosa compared with intrapatient noninflamed control tissue (38.6% vs 6.5%, p = 0.029). In cultured sinonasal epithelial cells from AFRS nasal polyps, Aspergillus fumigatus and Alternaria alternata stimulated higher SCC frequency compared with controls (27.4% vs 10.6%, p = 0.002; 18.1% vs 10.6%, p = 0.046), which led to increased IL-25 secretion in culture media (75.5 vs 3.3 pg/mL, p < 0.001; 32.3 vs 3.3 pg/mL, p = 0.007). Ki-67 expression was higher in SCCs grown in fungal stimulation conditions compared with controls. Although fungal antigens are known to potentiate immune response through innate cytokines, including IL-25, the early expansion of SCCs in the presence of fungus has not been described. This early event in the pathogenesis of noninvasive fungal rhinosinusitis may represent a target for intervention.

Effects of crude extracts of &lt;i&gt;Aspergillus fumigatus&lt;/i&gt; and &lt;i&gt;Penicillium citrinum&lt;/i&gt; on the biomass of &lt;i&gt;Eleusine indica&lt;/i&gt; (L.) Gaertn

In a pot experiment, two fungal strains namely Aspergillus fumigatus and Penicillium citrinum were evaluated for their herbicidal properties on the biomass of Eleusine indica using the biomass reduction method. Crude extracts of Aspergillus fumigatus and Penicillium citrinum made up of 10 treatments at three levels each with positive and negative controls in three replications in a Completely Randomised Block Design was adopted. Infections were evident within 48 hours after the application of the various treatments. At 1, 3, 5 and 7 days after application of the treatments, the percentage biomass yields and biomass reductions of E. indica were determined. The data collected were subjected to analysis of variance and the means separated using Duncan Multiple Range Test. The percentage reductions in biomass yields (88.46% and 35.61%) of Eleusine indica especially at the highest inocula concentration (1010 spores/ml) of the crude extracts showed significant differences (p≤ 0.001). The results indicate that both crude extracts possess the potentials for herbicidal control of Eleusine indica with evidence that the concentrated crude extracts of Aspergilus fumigatus had the greatest impact on its biomass yields. Therefore, it is recommended that further work be carried out on the purification of both crude extracts to confirm the effective biological metabolites accountable for their herbicidal properties and also evaluate these herbicidal properties on other weed species adopting different time and methods of application. Keywords: Fungal strains, Herbicidal properties, Biological metabolites, Concentrated crude extracts, Biomass yields and Biomass reductions

Priming with Toll-like receptor 3 agonist or interferon-gamma enhances the therapeutic effects of human mesenchymal stem cells in a murine model of atopic dermatitis

BackgroundAtopic dermatitis (AD) is a chronic and relapsing inflammatory skin disease. Great efforts have been recently made to treat AD using mesenchymal stem cells (MSCs), which have immunomodulatory functions. However, the immunomodulatory effects of MSCs need to be enhanced for clinical application in the treatment of AD.ObjectivesTo evaluate and characterise the therapeutic effects of human Wharton’s jelly-derived MSCs (WJ-MSCs) primed with the Toll-like receptor 3 agonist poly I:C or interferon-γ (IFN-γ) in a murine model of AD.MethodsMice were treated with Aspergillus fumigatus extract to induce AD and then subcutaneously injected with non-primed, poly I:C-primed or IFN-γ-primed WJ-MSCs. Clinical symptom scores, transepidermal water loss (TEWL), histological characteristics and cytokine levels were determined. Transcriptome profiling and pathway analyses of primed WJ-MSCs were conducted.ResultsThe clinical symptom score and TEWL in skin lesions were reduced in mice administered non-primed and primed WJ-MSCs. Epidermal thickness and inflammatory cell infiltration in skin lesions were reduced more in mice administered primed WJ-MSCs than in mice administered non-primed WJ-MSCs. Secretion of interleukin-17 was significantly reduced in skin draining lymph nodes of mice administered primed WJ-MSCs. Genomics and bioinformatics analyses demonstrated the enrichment of certain pathways specifically in WJ-MSCs primed with poly I:C or IFN-γ.ConclusionsPriming with poly I:C- or IFN-γ improved the therapeutic effects of WJ-MSCs in a murine model of AD. This study suggests that priming with poly I:C or IFN-γ enhances the immunomodulatory functions of WJ-MSCs and can be used as a novel therapeutic approach for AD.

Human IgE mAbs define variability in commercial Aspergillus extract allergen composition.

Sensitization to Aspergillus species is associated with allergic respiratory diseases. Allergen immunotherapy with nonstandardized Aspergillus extracts is commonly used as therapy in these patients. Unfortunately, no method exists to measure the relevant allergen protein content in diagnostic and therapeutic extracts. Thus, there is a critical need for Aspergillus extract standardization. We hypothesized that development of Aspergillus-specific human IgE mAbs would allow for the characterization of the relevant human allergenic epitopes among currently available commercial Aspergillus fumigatus extracts. Patients with allergic bronchopulmonary mycosis were recruited from Vanderbilt University Medical Center. IgE antibody-secreting B cells were grown and immortalized using human hybridoma techniques first described here. Twenty-six human Aspergillus-reactive IgE mAbs were used as capture and detection reagents to characterize the Aspergillus allergen content of commercial extracts. We found extreme variability in the specificity and quantity of their protein targets. Just 4 mAbs reacted with all available extracts, and only 1 of 4 extracts contained the major allergen Asp f 1. This degree of variability will almost certainly affect the efficacy of these reagents when used in diagnosis and treatment. Human IgE mAbs represent an innovative tool for the evaluation of relevant human allergenic epitopes, which may assist in future development and long-term standardization of mold extracts.

A synthetic glycosaminoglycan reduces sinonasal inflammation in a murine model of chronic rhinosinusitis.

Chronic rhinosinusitis (CRS) is characterized by sustained mucosal inflammation, impaired mucociliary clearance, loss of cilia and epithelial barrier breakdown, and tissue remodeling. Certain glycosaminoglycans inhibit various inflammatory mediators, suppress bacterial growth, and provide important functions in mucosal tissue repair and mucociliary clearance. Herein, we evaluated the effects of a synthetic glycosaminoglycan, GM-1111, on the clinical signs and inflammatory tissue changes associated with CRS in mice. CRS was generated by repeated intranasal applications of Aspergillus fumigatus (A. fumigatus) extracts over 4 weeks. Mice were then intranasally administered GM-1111 (600 μg per dose, 5 times a week) or vehicle (phosphate buffered saline, PBS) for an additional 4 weeks while still being given A. fumigatus extracts to maintain a chronic inflammatory environment with acute exacerbations. Clinical signs indicative of sinonasal inflammation were recorded throughout the study. After 9 weeks, whole blood and sinonasal tissues were harvested for hematological, histological, and biochemical examination. The clinical signs, white blood cell counts, tissue markers of sinonasal inflammation, and histological changes caused by A. fumigatus extract administration were compared to the healthy (PBS vehicle) and GM-1111-treated groups (n = 12 per treatment group). Compared to vehicle-treated animals, animals treated with GM-1111 demonstrated significant reductions in clinical signs (p<0.05), degenerative tissue changes, goblet cell hyperplasia, inflammatory cell infiltration (p<0.01), innate immunity- (tlr2, tlr4, myd88, il1b, tnfa, il6, and il12) and adaptive immunity-associated (ccl11, ccl24, ccl5, il4, il5, and il13) cytokine gene expression (p<0.05 to p<0.0001) in sinonasal tissues, and serum IgE levels (p<0.01). Our data suggest that GM-1111 significantly reduces local and systemic effects of CRS-associated sinonasal inflammation.

C3a receptor antagonism as a novel therapeutic target for chronic rhinosinusitis.

Chronic rhinosinusitis with nasal polyps (CRSwNP) is an inflammatory disease with an unknown etiology. Recent studies have implicated the complement system as a potential modulator of disease immunopathology. We preformed proteomic pathway enrichment analysis of differentially increased proteins, and found an enrichment of complement cascade pathways in the nasal mucus of individuals with CRSwNP as compared to control subjects. Sinonasal mucus levels of C3 correlated with worse subjective disease severity, whereas no significant difference in systemic C3 levels could be determined in plasma samples. Given that human sinonasal epithelial cells were the predominate sinonasal source of C3 and C3a staining, we focused on their role in in-vitro studies. Baseline intracellular C3 levels were higher in CRSwNP cells, and following exposure to Aspergillus fumigatus (Af) extract they released significantly more C3 and C3a. Inhibition of C3aR signaling led to a decrease in Af-induced C3 and C3a release, both in-vitro and in-vivo. Finally, we found in-vivo that C3aR deficiency or inhibition significantly reduced inflammation and CRS development in a mouse model of Af induced CRS. These findings demonstrate that local sinonasal complement activation correlates with subjective disease severity, and that local C3aR antagonism significantly ameliorates Af induced CRS in a rodent model.