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Background: Candida albicans and similar species are significant pathogens in immunocompromised and hospitalized individuals, known for mucosal colonization and bloodstream/organ invasion. Many pathogenic fungi, including these species, exhibit autofluorescence (R/OF) under specific light conditions, a feature crucial for their detection., Aim: We investigated the use of a 405 nm diode laser for the direct observation of red/orange autofluorescence of Candida spp., common in the oral cavity, exploring its potential in health screenings., Methods: This study utilized cultures of Candida spp. on Sabouraud dextrose agar with Qdot 655 and 685 for fluorescence benchmarking, illuminated using a 405 nm diode laser (continuous wave, power 250 mW, 0.0425 J/cm² fluence, 0.0014 W/cm² power density). Images were captured using a yellow-filter camera at set intervals (48 to 144 h). Visual and computational analyses evaluated the R/OF in terms of presence, intensity, coloration, and intra-colony variation., Results: Most Candida strains displayed red/orange autofluorescence at all observation times, characterized by varied coloration and intra-colony distribution. Initially, there was an increase in R/OF intensity, which then stabilized in the later stages of observation., Conclusions: The majority of the Candida strains tested are capable of emitting R/OF under 405 nm laser light. This finding opens up new possibilities for integrating R/OF detection into routine dental screenings for Candida spp.

Bacillus anthracis lethal toxin and edema toxin are binary toxins that consist of a common cell-binding moiety, protective antigen (PA), and the enzymatic moieties, lethal factor (LF) and edema factor (EF). PA binds to either of two receptors, capillary morphogenesis protein-2 (CMG-2) or tumor endothelial marker-8 (TEM-8), which triggers the binding and cytoplasmic translocation of LF and EF. However, the distribution of functional TEM-8 and CMG-2 receptors during anthrax toxin intoxication in animals has not been fully elucidated. Herein, we describe an assay to image anthrax toxin intoxication in animals, and we use it to visualize TEM-8- and CMG-2-dependent intoxication in mice. Specifically, we generated a chimeric protein consisting of the N-terminal domain of LF fused to a nuclear localization signal-tagged Cre recombinase (LFn-NLS-Cre). When PA and LFn-NLS-Cre were coadministered to transgenic mice expressing a red fluorescent protein in the absence of Cre and a green fluorescent protein in the presence of Cre, intoxication could be visualized at single-cell resolution by confocal microscopy or flow cytometry. Using this assay, we found that: (a) CMG-2 is critical for intoxication in the liver and heart, (b) TEM-8 is required for intoxication in the kidney and spleen, (c) CMG-2 and TEM-8 are redundant for intoxication of some organs, (d) combined loss of CMG-2 and TEM-8 completely abolishes intoxication, and (e) CMG-2 is the dominant receptor on leukocytes. The novel assay will be useful for basic and clinical/translational studies of Bacillus anthracis infection and for clinical development of reengineered toxin variants for cancer treatment., Competing Interests: Conflict of interest The authors declare no competing financial interests in relation to the work described., (Copyright © 2021. Published by Elsevier Inc.)

The orphan transcription factor nuclear receptor-related 1 protein (Nurr1, also known as NR4A2) plays a key role in embryonic development and maintenance of mesencephalic dopaminergic neurons in the substantia nigra. Nurr1 deficiency is associated with Parkinson's disease where dopaminergic neurons degenerate suggesting that counter-regulation of Nurr1 activity may have therapeutic effects. Here, we bacterially expressed and isolated a human Nurr1 fusion protein containing a N-terminal cell delivery domain derived from detoxified anthrax lethal factor followed by wild type ubiquitin with deubiquitinating enzyme recognition site for intracellular cleavage. Addition of the Nurr1 fusion protein to dopaminergic SH-SY5Y cells generated a cleaved, cytosolic Nurr1-containing fragment which was associated with increased levels of tyrosine hydroxylase, the rate-limiting enzyme in dopamine synthesis. Promoter-activity assays confirmed that exposure of cells to full-length Nurr1 fusion protein activated not only its cognate human tyrosine hydroxylase promoter but also the corresponding mouse sequence, although at a reduced efficiency. Using 6-hydroxydopamine as a dopaminergic cell specific neurotoxin, we demonstrate that full-length Nurr1 fusion protein promotes a concentration-dependent protection from this toxic insult. Altogether, the enhancement of tyrosine hydroxylase in naïve dopaminergic cells and the protective effects in a cellular model of Parkinson's disease suggest that full-length Nurr1 fusion protein may contribute to the development of a novel concept of protein-based therapy.