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      Detection of glucosamine as a marker for Aspergillus niger: a potential screening method for fungal infections

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          Graphical abstract

          Several species of fungus from the genus Aspergillus are implicated in pulmonary infections in immunocompromised patients. Broad screening methods for fungal infections are desirable, as cultures require a considerable amount of time to provide results. Herein, we developed degradation and detection methods to produce and detect D-glucosamine (GlcN) from Aspergillus niger, a species of filamentous fungus . Ultimately, these techniques hold the potential to contribute to the diagnosis of pulmonary fungal infections in immunocompromised patients. In the following studies, we produced GlcN from fungal-derived chitin to serve as a marker for Aspergillus niger. To accomplish this, A. niger cells were lysed and subjected to a hydrochloric acid degradation protocol. Products were isolated, reconstituted in aqueous solutions, and analyzed using hydrophilic interaction liquid chromatography (HILIC) in tandem with electrospray ionization time-of-flight mass spectrometry. Our results indicated that GlcN was produced from A. niger. To validate these results, products obtained via fungal degradation were compared to products obtained from the degradation of two chitin polymers. The observed retention times and mass spectral extractions provided a two-step validation confirming that GlcN was produced from fungal-derived chitin. Our studies qualitatively illustrate that GlcN can be produced from A. niger; applying these methods to a more diverse range of fungi offers the potential to render a broad screening method for fungal detection pertinent to diagnosis of fungal infections.

          Supplementary Information

          The online version contains supplementary material available at 10.1007/s00216-021-03225-7.

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          Nosocomial candidemia: risk factors and attributable mortality.

          Over the past decade, the incidence of hospital-acquired bloodstream infections caused by Candida species has risen and the species associated with such infections have changed. The incidence of candidemia is dramatically higher in high-risk, critical-care units than in other parts of the hospital. Certain underlying physical conditions including acute leukemia, leukopenia, burns, gastrointestinal disease, and premature birth predispose patients to nosocomial candidemia. Independent risk factors include prior treatment with multiple antibiotics, prior Hickman catheterization, isolation of Candida species from sites other than the blood, and prior hemodialysis. In this article some of the challenges posed by the management of nosocomial candidemia are presented in three case studies. In addition, the results of several investigations of nosocomial candidemia at the University of Iowa Hospitals and Clinics are reviewed.
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            Hyphal differentiation in the exploring mycelium of Aspergillus niger.

            Mycelial fungi play a central role in element cycling in nature by degrading dead organic material such as wood. Fungal colonization of a substrate starts with the invasion of exploring hyphae. These hyphae secrete enzymes that convert the organic material into small molecules that can be taken up by the fungus to serve as nutrients. Using green fluorescent protein (GFP) as a reporter, we show for the first time that exploring hyphae of Aspergillus niger differentiate with respect to enzyme secretion; some strongly express the glucoamylase gene glaA, while others hardly express it at all. When a cytoplasmic GFP was used, 27% of the exploring hyphae of a 5-day-old colony belonged to the low expressing hyphae. By fusing GFP to glucoamylase and by introducing an ER retention signal, this number increased to 50%. This difference is due to cytoplasmic streaming of the reporter in the former case, as was shown by using a photo-activatable GFP. Our findings indicate that a fungal mycelium is highly differentiated, especially when taking into account that hyphae in the exploration zone were exposed to the same nutritional conditions.
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              Fungal chitosan production and its characterization.

              The objective of this investigation was to evaluate the chitosans produced by several species of fungi. Representatives of four species of filamentous fungi, Aspergillus niger, Rhizopus oryzae, Lentinus edodes and Pleurotus sajo-caju, and two yeast strains, Zygosaccharomyces rouxii TISTR5058 and Candida albicans TISTR5239, were investigated for their ability to produce chitosan in complex media. Fungal chitosan was produced at 10-140 mg g-1 cell dry weight, had a degree of deacetylation of 84-90% and a molecular weight of 2.7 x 104-1.9 x 105 Da with a viscosity of 3.1-6.2 centipoises (cP). Rhizopus oryzae TISTR3189 was found to be the producer of the highest amounts of chitosan. Commercial chitosan could be obtained from Rhizopus mycelia and would have potential applications for medical and agricultural uses.
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                Author and article information

                Contributors
                Melissa.reynolds@colostate.edu
                Journal
                Anal Bioanal Chem
                Anal Bioanal Chem
                Analytical and Bioanalytical Chemistry
                Springer Berlin Heidelberg (Berlin/Heidelberg )
                1618-2642
                1618-2650
                22 February 2021
                22 February 2021
                2021
                : 413
                : 11
                : 2933-2941
                Affiliations
                [1 ]GRID grid.47894.36, ISNI 0000 0004 1936 8083, Department of Chemistry, , Colorado State University, ; 1801 Campus Delivery, Fort Collins, CO 80523 USA
                [2 ]GRID grid.47894.36, ISNI 0000 0004 1936 8083, Department of Microbiology, Immunology, and Pathology, , Colorado State University, ; 1601 Campus Delivery, Fort Collins, CO 80523 USA
                [3 ]GRID grid.47894.36, ISNI 0000 0004 1936 8083, Department of Chemical and Biological Engineering, , Colorado State University, ; 1370 Campus Delivery, Fort Collins, CO 80523 USA
                [4 ]GRID grid.47894.36, ISNI 0000 0004 1936 8083, School of Biomedical Engineering, , Colorado State University, ; 1376 Campus Delivery, Fort Collins, CO 80523 USA
                Article
                3225
                10.1007/s00216-021-03225-7
                8043943
                33615396
                89cf2ed8-0ea7-4d1d-816f-e512b704138b
                © The Author(s) 2021

                Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

                History
                : 7 December 2020
                : 21 January 2021
                : 8 February 2021
                Funding
                Funded by: FundRef http://dx.doi.org/10.13039/100000009, Foundation for the National Institutes of Health;
                Award ID: 5R01HL140301-02
                Categories
                Research Paper
                Custom metadata
                © Springer-Verlag GmbH Germany, part of Springer Nature 2021

                Analytical chemistry
                aspergillus,lc-ms,chitin,glucosamine
                Analytical chemistry
                aspergillus, lc-ms, chitin, glucosamine

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