Mycopathologia最新文献

Aspergillus fumigatus, an opportunistic and allergenic pathogenic fungus, is responsible for a range of clinical disorders in humans, including invasive aspergillosis (IA), which can lead to severe infections in immunocompromised individuals. Unfortunately, the emergence of azole resistance has become a significant challenge in combating IA, necessitating further investigations into the underlying mechanisms of resistance. In this study, we conducted an integrated proteomic and phosphoproteomic analysis of biofilm proteins from both azole-resistant and wildtype strains of A. fumigatus under voriconazole pressure. Our proteomic analysis identified 148 upregulated and 146 downregulated proteins in the azole-resistant strains, while phosphoproteomic analysis revealed 316 upregulated phosphopeptides and 109 downregulated phosphopeptides, suggesting extensive phosphorylation modifications associated with azole resistance. Upon excluding the impact of protein changes, we identified 133 proteins with differential expression solely at the phosphorylation level, comprising 104 upregulated and 29 downregulated proteins. Functional annotation and analysis highlighted the significance of these differentially expressed phosphoproteins in cell wall integrity, filamentous growth, and high-osmolarity stress response, with 33 MAPK pathway-associated proteins displaying phosphopeptide level regulation. These findings provide valuable insights into the mechanisms behind azole resistance in A. fumigatus and offer potential new drug targets for combating this pathogenic fungus in humans.

This study evaluates and compares the diagnostic and prognostic capabilities of ChatGPT-4o and DeepSeek-R1 in 56 HIV-negative talaromycosis cases. Clinical case fragments were de-identified and submitted to both models, with diagnostic accuracy and prognostic prediction rates statistically analyzed using chi-square tests, Fisher's exact tests, and logistic regression. Results showed DeepSeek-R1 achieved significantly higher diagnostic accuracy (66.1%) than ChatGPT-4o (3.6%) (χ² = 48.2, p < 0.001), attributable to its regional data training focusing on Southeast Asia and southern China. Conversely, ChatGPT-4o demonstrated superior prognostic prediction accuracy (78.6% vs. 50.0%, p < 0.001), with 90.2% specificity for improved (survival) outcomes, while DeepSeek-R1 showed 86.7% sensitivity for mortality. Key diagnostic predictors included hilar lymphadenectasis (odds ratio [OR] = 6.8, 95% confidence interval [CI]: 2.1-22.3, P = 0.002) and chest pain (OR = 5.9, 95% CI: 1.4-25.6, P = 0.016). The findings highlight DeepSeek-R1's regional diagnostic advantage and ChatGPT-4o's prognostic utility, advocating for their collaborative use to enhance early detection and management of this neglected fungal infection in immunocompromised, non-HIV populations.

Background: Lower respiratory tract fungal infections (LRTFIs) contribute significantly to global disease burden, yet systematic research is limited. This study analyzes their disability-adjusted life year (DALY) burden, exploring trends, geographic patterns, demographic differences, driving factors, and inequalities.

Methods: Using Global Burden of Disease Study 2021 data, we analyzed global, regional, and national DALYs trends from 1990 to 2021 and forecasted burden to 2050. Decomposition analysis evaluated impacts of population growth, ageing, and epidemiological changes. We analyzed national burden inequalities using the Slope Index of Inequality and the Concentration Index.

Results: Global DALYs from LRTFIs decreased slightly from 1.32 to 1.30 million (1990-2021), with an age-standardized DALY rate (ASDR) decline from 25.86 to 16.37 per 100,000 (EAPC = - 1.52). The highest ASDRs were in low-SDI regions, particularly Central Sub-Saharan Africa (71.86 per 100,000). Zimbabwe, Lesotho, and the Central African Republic had the highest national burdens. Males and individuals over 50 had higher DALY rates. Population growth increased DALYs, while ageing and epidemiological shifts reduced then. Absolute inequality declined, but relative inequality increased. By 2050, DALYs are projected to rise to 1.78 million, though ASDR will decline to 10.37 per 100,000.

Conclusions: Despite progress, LRTFIs burden remains high in underdeveloped regions, particularly Africa. Population growth and ageing will drive future challenges, and significant global inequalities in disease burden persist.

Aspergillus section Flavi comprises opportunistic pathogens such as Aspergillus flavus, posing significant health risks. Unlike A. fumigatus, where environmental selection drives widespread resistance, it is uncertain whether A. flavus develops azole resistance through a similar environmental route. This study analyzed 544 Aspergillus section Flavi isolates collected from 534 patients in Dutch hospitals (1994-2023). Calmodulin sequencing was used for molecular identification. Antifungal susceptibility testing (AFST) was performed according to EUCAST (European Committee on Antimicrobial Susceptibility Testing) guidelines. Cyp51A was sequenced and mutations were mapped onto the CYP 51 protein 3D model for azole-resistant or non-wild-type isolates. Clinical data, including azole exposure history and underlying diseases, were correlated with the resistance profiles. Of the 544 isolates, 520 were identified as A. flavus and 24 as related species, including A. tamarii (16), A. parasiticus (3), A. nomiae (2), A. pseudonomiae (2), and A. pseudocaelatus (1). Fourteen isolates from five patients were azole-resistant/non-WT, with resistance rates ranging from 0.2 to 0.8%. Resistant/non-WT isolates were associated with chronic diseases and prior clinical azole exposure. Among 10 patients with proven/probable invasive aspergillosis azole resistance was not observed. Two main Cyp51A-SNPs were found in azole-resistant isolates: Y119F and T329A. Aspergillus flavus was the dominant pathogenic species within the section Flavi with low levels of azole resistance. Distribution of resistant cases supports in host resistance selection rather than environmental selection. These findings highlight the need for further research on the ecological and molecular factors that influence resistance in A. flavus.

Abdominal aortic aneurysm (AAA) is a life-threatening vascular condition characterized by inflammatory degeneration of the vessel wall. Emerging evidence suggests that microbial factors contribute to its progression. In this study, we analyzed the mycobiome composition of stool, blood, thrombus and damaged vessel wall samples collected during surgery from 24 AAA patients using Internal Transcribed Spacer (ITS) sequencing. Significant differences in alpha and beta diversity were observed across the sample types, confirming compartmentalization of the mycobiome. However, individual fungal profiles did not establish a clear gut-blood-vessel wall axis, indicating that fungi translocated to the vessel wall may originate from other anatomical regions. Comparison AAA mycobiome with healthy arterial walls' mycobiome from organ donors revealed a dominance of anti-inflammatory Tomentella in healthy samples, while pro-inflammatory Malassezia species were prevalent in damaged vessel walls. These findings highlight the role of fungi in AAA progression and suggest potential thera-peutic avenues, including antifungal adjuvant treatments to mitigate inflammation and aneurysm development.

Candidiasis, traditionally dominated by Candida albicans infection, now faces a paradigm shift with the emergence of non-C. albicans infections, such as the multidrug-resistant (MDR) Candidozyma (formerly Candida) species. Critically, the emergence of multidrug resistance (MDR) Candidozyma duobushaemulonii (also known as Candida duobushaemulonii) poses critical challenges to antifungal therapy. Here, in this comparative study, we analyzed four clinical C. duobushaemulonii isolates (cd, cd1, cd2, and cd3) to establish strain-specific virulence and antifungal resistance profiles. Biofilm quantification using crystal violet (CV) and 2,3-Bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT) metabolic activity assays showed different biomass production levels across strains. Cellular aggregation capacity differed significantly, while cell surface hydrophobicity (CSH) showed inverse patterns. In Galleria mellonella infection models, virulence stratification was observed. Antifungal susceptibility testing revealed minimum inhibitory concentration (MIC) azole gradients. MDR in the cd1 strain extended to 5-fluorocytosine (5-FC) and anidulafungin (ANI), whereas the cd3 strain showed intermediate posaconazole (PSZ) resistance distinct to the cd2 strain. Amino acid sequence analyses indicated that Y132F substitutes in ERG11 and D139N in FUR1 genes occurred in cd and cd1 strains. Gene expression analysis recorded various regulatory situations among strains. These analyses provide a complete resistance-virulence matrix for C. duobushaemulonii, establishing baseline characteristics for epidemiological surveillance and a reference framework for future comparative studies.

Scedosporium species are opportunistic pathogens causing a large variety of human infections. To date, there is limited information on the pathogenic mechanisms of these fungi, partly because of the limited number of genetic tools available. Here, the CRISPR-Cas9 technology, which provided promising results for functional genomic studies in filamentous fungi, was optimized for Scedosporium species using in vitro assembled Cas9 ribonucleoprotein (RNP) complexes. In these fungi, functional genomic studies are particularly complex in a wild-type strain, because of the high frequency of non-homologous recombination. Prior disruption of the KU70 gene encoding one of the components of the non-homologous end joining system is required, which necessitates the use of a first selection marker. The cleavage of the target gene at each end using a dual RNA-guided Cas9 complex, followed by recombination with a repair template containing the hygromycin resistance gene, allowed disruption of the target gene in the ΔKU70 mutant. Four genes encoding dioxygenases, catalyzing the critical ring-opening step in aromatic hydrocarbons, were successfully disrupted, and the optimum efficiency was observed using 5 μg of the HygR repair cassette. Alternatively, in the wild-type strain, the exclusive use of two Cas9 RNP complexes was enough to achieve an efficient deletion method; one dioxygenase gene was successfully deleted in up to 20% of the obtained colonies. These last experimental conditions path the way to multiple gene deletions and complementation experiments, which cannot be reached using our first procedure since only two selection markers are available for Scedosporium species.

Introduction: Invasive pulmonary aspergillosis (IPA) in hematological malignancy populations has high mortality rates. While there are identified prognostic factors for mortality, conflicting results are reported from the studies including bronchoalveolar lavage galactomannan (GM) measurements and co-infections. We aimed to evaluate risk factors associated with in-hospital mortality in a hematological malignancy population undergoing bronchoscopy with a preliminary diagnosis of IPA in a single tertiary care center.

Method: Patients undergoing bronchoscopy with a preliminary diagnosis of IPA were included in this retrospective study. Bacterial co-infection was defined as a positive bacterial culture in respiratory samples within ± 7 days of the index bronchoscopy procedure.

Results: Study population consisted 305 patients diagnosed as possible, probable or proven IPA. 57 patients presented with fungal and bacterial co-infection. In-hospital mortality was observed in 98 (33.6%) patients. Patients with mortality status were older and were not in remission for hematological malignancy. Serum GM, bronchoalveolar lavage (BAL) GM and bronchial lavage (BL) GM levels showed a significant relation with in-hospital mortality but weak accuracy. In multivariate analysis for risk factors of in-hospital mortality; age, remission status, number of nodules in HRCT and having fungal infections with or without combined bacterial infection were independent risk factors.

Conclusion: Co-infections in this susceptible population should not be overlooked even in the presence of fungal infection evidence. BAL and BL GM tended to have a stronger relation with survival than serum GM measurements taken at the same time. The number of nodules in radiological assessment might be an indicator of mortality. Radiological changes through the disease course might be assessed in further studies.

Background and aims: Invasive cutaneous fungal infections (ICFIs) are life-threatening complications in pediatric cancer patients. This study aimed to investigate the prevalence, clinical characteristics, and outcomes of ICFIs among pediatric cancer patients hospitalized at a referral oncology teaching Hospital in Shiraz, Iran.

Methods: This cross-sectional study included pediatric patients with malignancies and suspected ICFIs who were admitted to the Amir Oncology Teaching Hospital between 2015 and 2022. Diagnosis was based on the EORTC/MSG criteria and confirmed using clinical, microbiological, and histopathological methods. Data were analyzed using descriptive statistics, Kaplan-Meier survival analysis, and comparative tests, with a significance threshold of p < 0.05.

Results: Among the 24 patients, 58.3% were boys, and 45.8% were aged 1-5 years. Acute lymphoblastic leukemia was the most common malignancy (27.3%). Mucoralean fungi (36.4%) and Aspergillus (59.1%) were the most common. The overall survival rate was 54.2%. Proven ICFIs had the poorest outcomes, with a survival probability declining to zero by month 26. Patients with lower CRP levels and febrile neutropenia had better outcomes (p < 0.001 and p = 0.041, respectively), but survival rates did not vary significantly according to sex, age, or treatment approach (monotherapy versus combination therapy).

Conclusion: In pediatric oncology patients, ICFIs are associated with high mortality, particularly in cases of mucormycosis or proven infections. Improving survival depends on early diagnosis, risk stratification, and rapid management, particularly in patients with neutropenia and fever.