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Journal of Clinical Immunology Apr 2020Autosomal recessive (AR) CARD9 deficiency is an inherited immune disorder which results in impaired innate immunity against various fungi. Superficial and invasive... (Review)
Review
PURPOSE
Autosomal recessive (AR) CARD9 deficiency is an inherited immune disorder which results in impaired innate immunity against various fungi. Superficial and invasive fungal infections, mainly caused by Candida or Trichophyton species, are the hallmark of CARD9 deficiency. Together with the increasing number of CARD9-deficient patients reported, different pathogenic fungal species have been described such as Phialophora, Exophiala, Corynespora, Aureobasidium, and Ochroconis. Saprochaete capitata is an opportunistic infectious agent in immunocompromised patients and is a common cause of invasive fungal disease in patients with hematological malignancies. In this study, we investigated the causative genetic defect in a patient with S. capitata fungal infection which disseminated to lymph nodes and common bile duct.
METHODS
The identification of the isolated yeast strain was made by direct microscopic examination and confirmed by internal transcribed spacer (ITS) sequencing. We applied whole exome sequencing to search for the disease-causing mutation. Sanger sequencing was used to validate the mutation in the patient and his parents.
RESULTS
S. capitata was isolated from the biopsy specimen as the causative microorganism responsible for the invasive fungal disease in the patient. Whole exome sequencing revealed a homozygous c.883C > T, (p.Q295*) mutation in CARD9, confirmed by Sanger sequencing.
CONCLUSIONS
This is the first report of invasive Saprochaete infection associated with autosomal recessive (AR) CARD9 deficiency in the literature and thereby further extends the spectrum of fungal diseases seen in these patients.
Topics: Adolescent; CARD Signaling Adaptor Proteins; Candidiasis, Chronic Mucocutaneous; Cholestasis; Chromosome Disorders; Genes, Recessive; Humans; Immunocompromised Host; Invasive Fungal Infections; Iraq; Male; Saccharomycetales; Sequence Deletion; Exome Sequencing
PubMed: 32020378
DOI: 10.1007/s10875-020-00759-w -
Journal of Agricultural and Food... Feb 2023Zearalenone (ZEN) is an estrogenic mycotoxin most frequently found in cereals that can cause reproductive disorders in livestock and pose a severe threat to animal...
Zearalenone (ZEN) is an estrogenic mycotoxin most frequently found in cereals that can cause reproductive disorders in livestock and pose a severe threat to animal husbandry. In this study, we isolated a ZEN-degrading strain from soil and found that ZenH, a hydrolase, is responsible for the hydrolysis of ZEN through comparative proteomics and biochemical studies. ZenH exhibited the highest similarity with lactone hydrolase ZHD607 from at 21.52%. ZenH displayed maximal enzymatic activity at pH 7.0 and 55 °C with a Michaelis constant of 12.64 μM. The catalytic triad of ZenH was identified as S117-D142-H292 by molecular docking and site-directed mutagenesis. ZenH catalyzed the hydrolysis of ZEN to a novel metabolite, (,)-4-hydroxy-2-(10-hydroxy-6-oxoundec-1-en-1-yl)-7-oxabicyclo[4.2.0]octa-1,3,5-trien-8-one, which exhibited significantly lower estrogenic toxicity than ZEN. This study illustrates a novel ZEN-degrading enzyme and reveals a new degradation product. Furthermore, the enzyme showed good potential for detoxifying ZEN during food processing.
Topics: Animals; Zearalenone; Hydrolases; Molecular Docking Simulation; Mycotoxins; Biodegradation, Environmental
PubMed: 36701132
DOI: 10.1021/acs.jafc.2c06410 -
Microorganisms Apr 2020Dark septate endophytes (DSEs) are often trace element (TE)-tolerant fungi and are abundant in TE-polluted environments. The production of melanin, a black polymer found...
Dark septate endophytes (DSEs) are often trace element (TE)-tolerant fungi and are abundant in TE-polluted environments. The production of melanin, a black polymer found in cell walls, was hypothesized by several authors to play a role in the TE tolerance of DSEs. To test this hypothesis, we established a series of experiments using albino strains and melanin inhibitors and examined the responses to Cd and Zn. Six DSEs belonging to genera sp., sp. and , were evaluated. The strains mainly produced 1,8-dihydroxynaphthalene (DHN) melanin whereas 3,4-dihydroxyphenylalanin melanin was also synthetized. Cd and Zn decreased melanin synthesis in most of the strains. A reduction in melanin concentration in hyphae through the use of tricyclazole, an inhibitor of DHN-melanin synthesis, did not reduce the tolerance of the strains to Cd and Zn. Similarly, albino mutants of sp. were not more sensitive to Cd and Zn than the WT strain. Moreover, tricyclazole-treated colonies accumulated less Cd but more Zn compared to untreated colonies. The Cd and Zn contents of albino strains were variable and similar to that of the WT. The results suggest that melanin production is not an important functional trait that contributes to Cd and Zn tolerance, but might contribute to Cd accumulation.
PubMed: 32276491
DOI: 10.3390/microorganisms8040537 -
Antimicrobial Agents and Chemotherapy Sep 2014The in vitro activities of nine antifungal drugs and their combinations against 31 clinical and 15 environmental Phialophora verrucosa strains were tested. The MIC90/90%...
The in vitro activities of nine antifungal drugs and their combinations against 31 clinical and 15 environmental Phialophora verrucosa strains were tested. The MIC90/90% minimum effective concentration (MIC/MEC90) values (μg/ml) across all strains were as follows: for terbinafine, 0.25; for posaconazole, 0.5; for voriconazole, 1; for itraconazole, 2; for amphotericin B, 4; for caspofungin and micafungin, 16; and for fluconazole and flucytosine, 64. The highest synergy was shown by the combination of itraconazole plus caspofungin (with synergy against 100% of the 31 clinical strains), followed by amphotericin B plus flucytosine (45.2%) and itraconazole plus terbinafine or micafungin (25.8% or 12.9%, respectively).
Topics: Antifungal Agents; Chromoblastomycosis; Drug Combinations; Microbial Sensitivity Tests; Phialophora
PubMed: 24982078
DOI: 10.1128/AAC.02875-14 -
[Zhonghua Yan Ke Za Zhi] Chinese... Oct 2017To evaluate the clinical features of retrocorneal fungal infection and the therapeutic effects. This was a retrospective, noncomparative study of nine patients with...
To evaluate the clinical features of retrocorneal fungal infection and the therapeutic effects. This was a retrospective, noncomparative study of nine patients with retrocorneal fungal infection and an intact corneal epithelium treated at Qingdao Eye Hospital. The history, clinical features, diagnostic methods, pathogens and therapeutic effects were analyzed. Five patients had a history of trauma by plant, two had a non-plant injury, and two had unidentified reasons. The duration between the initial onset and the first visit to our hospital was 7.9 months (range, 3 months to 2 years). There was endothelial plaque and sometimes with white infiltration in the deep stroma, but the corneal epithelium remained integrated, and the anterior stroma was uninfected. The patients were misdiagnosed as uveitis, herpes simplex keratitis or bacterial keratitis in the other hospitals. Visual acuity was 20/200 in four eyes, 20/60 in one eye, 20/40 in two eyes, and 20/30 in two eyes. Fungal hyphae were detected by confocal microscopy in six eyes. All the eyes had poor response to the antifungal medication before penetrating keratoplasty was performed. The smear examinations of the corneal endothelial plaque showed fungal hyphae in six eyes. Alternaria Nees, Apospory, Phialophora verrucosa, and Fusarium were identified. Plant injury is the most common risk factor of retrocorneal fungal infection. Slow onset and no initially obvious symptoms may lead to delayed diagnosis and misdiagnosis. The diagnosis can be confirmed by confocal microscopy before surgery. The effect of antifungal medication is usually poor. .
Topics: Antifungal Agents; Eye Infections, Fungal; Humans; Keratoplasty, Penetrating; Mycoses; Retrospective Studies
PubMed: 29050189
DOI: 10.3760/cma.j.issn.0412-4081.2017.10.008 -
Medical Mycology Feb 2015In a retrospective study, we investigated 52 formalin-fixed, paraffin-embedded (FFPE) samples from cats with histologically confirmed cutaneous and subcutaneous mycoses...
In a retrospective study, we investigated 52 formalin-fixed, paraffin-embedded (FFPE) samples from cats with histologically confirmed cutaneous and subcutaneous mycoses to determine if the pathogens could be identified by molecular methods. Aim of the study was to obtain a deep understanding of the spectrum of infectious agents, which, as we hypothesized, was not available by histopathology alone. Detection of feline and fungal DNA was achieved in 92.3% and 94.2% of the samples, respectively. Most of the subcutaneous infections in cats were caused by Alternaria spp. (63.5%), followed by Cryptococcus neoformans (7.7%), Histoplasma capsulatum (5.8%), Sporothrix spp. (3.8%), Aspergillus vitricola, Aureobasidium pullulans, Exophiala attenuata, Fusarium oxysporum, Lecythophora cateniformis, Microsporum canis, and Phialophora sp. (1.9% each). The results from molecular identification indicate that correct identifications of the fungal pathogens by histology alone were rarely possible. The spectrum of fungal pathogens identified after DNA extraction from FFPE samples was much broader than that expected by classical histopathology. This was especially noted in alternariosis in that the micromorphological pattern in tissue was misleading and could be confused with that of cryptococcosis. Due to different susceptibilities to antifungal agents, it is important to arrive at a definitive diagnosis, which might be possible by examination of the fungus recovered in culture and/or molecular methods, in addition to the histopathologic techniques.
Topics: Animals; Cat Diseases; Cats; Dermatomycoses; Fungi; Molecular Diagnostic Techniques; Pathology, Molecular; Retrospective Studies
PubMed: 25550386
DOI: 10.1093/mmy/myu082 -
Frontiers in Immunology 2022-related inherited immune disorders are a major risk factor for chronic disseminated fungal infection. In addition to pathogens of and dermatophytes, the environmental...
-related inherited immune disorders are a major risk factor for chronic disseminated fungal infection. In addition to pathogens of and dermatophytes, the environmental opportunists of the black yeast-like fungi are relatively frequent in this patient cohort. Particularly the genus is overrepresented. We investigated two isolates of a strain of residing in a patient, sampled with a period of ten years apart. Genomes, melanization and antifungal susceptibility of progenitor and derived strains were compared, and potential adaptation to the host habitat was investigated with proteomic techniques using post-translational modification as a proxy. Global lactylation analysis was performed using high accuracy nano-LC-MS/MS in combination with enrichment of lactylated peptides from digested cell lysates, and subsequent peptide identification. The genome of the derived isolate had accumulated 6945 SNPs, of which 31 were detected in CDS. A large number of identified proteins were significantly enriched, e.g. in melanin biosynthesis. A total of 636 lactylation sites on 420 lactylated proteins were identified, which contained in 26 types of modification motifs. Lysine lactylation (Kla) was found in 23 constituent proteins of the ribosome, indicating an impact of Kla in protein synthesis. Twelve lactylated proteins participated in pathogenicity. A protein-protein interaction (PPI) network analysis suggested that protein lactylations are widely distributed influencing various biological processes. Our findings reveal widespread roles for lysine lactylation in regulating metabolism and melanin biosynthesis in black fungi. Several large rearrangements and inversions were observed in the genome, but genomic changes could not be linked to adaptation or to known clinically relevant properties of progenitor to derived isolate; antifungal susceptibility had largely remained unaltered.
Topics: Antifungal Agents; CARD Signaling Adaptor Proteins; Humans; Immune System Diseases; Lysine; Melanins; Phialophora; Protein Processing, Post-Translational; Proteomics; Tandem Mass Spectrometry
PubMed: 36003392
DOI: 10.3389/fimmu.2022.966457 -
Mycopathologia Jun 2015Caspase recruitment domain-containing protein 9 (CARD9) is an adaptor molecule that is critical for NF-κB activation and forms a complex with B cell lymphoma 10 and...
Caspase recruitment domain-containing protein 9 (CARD9) is an adaptor molecule that is critical for NF-κB activation and forms a complex with B cell lymphoma 10 and mucosa-associated lymphoid tissue lymphoma translocation gene 1 that mediates C-type lectin receptors (CLRs)-triggered intracellular signaling during antifungal immunity. However, the role of CARD9 in the host defense against Phialophora verrucosa (P. verrucosa) infection remains to be elucidated. In the present study, we investigated the functions of polymorphonuclear neutrophils (PMNs) from patients with CARD9 deficiencies against P. verrucosa. By isolating PMNs from patients and healthy blood donors and subsequently challenging the cells with P. verrucosa, we demonstrated that, compared with healthy donors, CARD9-deficient PMNs exhibited defects in P. verrucosa killing and pro-inflammatory cytokine productions, which can be rescued in the presence of serum; however, the CARD9-deficient PMNs exhibited normal reactive oxygen species generation and phagocytotic ability. In conclusion, our results indicate that CARD9 is indispensable for P. verrucosa killing by PMNs, and serum opsonization acts as a CARD9-independent way, which could be a promising immunotherapy in the future.
Topics: CARD Signaling Adaptor Proteins; Cytokines; Humans; Microbial Viability; Mycoses; Neutrophils; Nitric Oxide; Phagocytosis; Phialophora
PubMed: 25790941
DOI: 10.1007/s11046-015-9877-2 -
Mycopathologia Jun 2021Phialophora verrucosa (P. verrucosa) is a pathogen that can cause chromoblastomycosis and phaeohyphomycosis. Recent evidence suggests that neutrophils can produce...
Phialophora verrucosa (P. verrucosa) is a pathogen that can cause chromoblastomycosis and phaeohyphomycosis. Recent evidence suggests that neutrophils can produce neutrophil extracellular traps (NETs) that can protect against invasive pathogens. As such, we herein explored the in vitro functional importance of P. verrucosa-induced NET formation. By assessing the co-localization of neutrophil elastase and DNA, we were able to confirm the formation of classical NETs entrapping P. verrucosa specimens. Sytox Green was then used to stain these NETs following neutrophil infection with P. verrucosa in order to quantify the formation of these extracellular structures. NET formation was induced upon neutrophil exposure to both live, UV-inactivated, and dead P. verrucosa fungi. The ability of these NETs to kill fungal hyphae and conidia was demonstrated through MTT and pouring plate assays, respectively. Overall, our results confirmed that P. verrucosa was able to trigger the production of NETs, suggesting that these extracellular structures may represent an important innate immune effector mechanism controlling physiological responses to P. verrucosa infection, thereby aiding in pathogen control during the acute phases of infection.
Topics: Extracellular Traps; Humans; Hyphae; Neutrophils; Phialophora
PubMed: 34013384
DOI: 10.1007/s11046-021-00554-0 -
Molecular Plant-microbe Interactions :... Oct 2018Brown stem rot, caused by the fungus Phialophora gregata, reduces soybean yield by up to 38%. Although three dominant resistance loci have been identified (Rbs1 to...
Brown stem rot, caused by the fungus Phialophora gregata, reduces soybean yield by up to 38%. Although three dominant resistance loci have been identified (Rbs1 to Rbs3), the gene networks responsible for pathogen recognition and defense remain unknown. Further, identification and characterization of resistant and susceptible germplasm remains difficult. We conducted RNA-Seq of infected and mock-infected leaf, stem, and root tissues of a resistant (PI 437970, Rbs3) and susceptible (Corsoy 79) genotype. Combining historical mapping data with genotype expression differences allowed us to identify a cluster of receptor-like proteins that are candidates for the Rbs3 resistance gene. Reads mapping to the Rbs3 locus were used to identify potential novel single-nucleotide polymorphisms within candidate genes that could improve phenotyping and breeding efficiency. Comparing responses to infection revealed little overlap in differential gene expression between genotypes or tissues. Gene networks associated with defense, DNA replication, and iron homeostasis are hallmarks of resistance to P. gregata. This novel research demonstrates the utility of combining contrasting genotypes, gene expression, and classical genetic studies to characterize complex disease resistance loci.
Topics: Base Sequence; Gene Expression Regulation, Plant; Phialophora; Plant Diseases; Plant Proteins; RNA, Plant; Glycine max
PubMed: 30004290
DOI: 10.1094/MPMI-01-18-0009-R