Candidemia represent a significant cause of morbidity and mortality in hospitalized patients. Mortality depends on the virulence and resistance profile of the organism. This study aimed to evaluate virulence factors in Candida species isolated from bloodstream infections and correlate them with antifungal resistance patterns and clinical outcomes. 100 Candida isolates from blood cultures were collected over one year. Virulence factors including hemolysin, phospholipase, and proteinase activities, along with biofilm formation, were assessed. Susceptibility to antifungal was assessed using the VITEK®2 system. Clinical data and outcomes were analysed in relation to virulence factors and resistance patterns. Non-albicans Candida species (NAC) predominated (90%) over C. albicans (10%). C. parapsilosis (32%) and C. tropicalis (30%) were the most common NAC species. Overall, hemolysin activity was detected in 79% of isolates, phospholipase in 51%, proteinase in 43%, and biofilm formation in 87%. C. auris and C. duobushaemulonii demonstrated the strongest virulence factor expression. Fluconazole resistance was observed in 22% of isolates, with C. auris showing 100% resistance. Isolates with stronger enzymatic activities and biofilm formation showed significantly higher antifungal resistance (P < 0.0001). The crude mortality rate was 27%, with significantly higher mortality in patients infected with resistant isolates and those expressing strong virulence factors (P < 0.05). NAC species have emerged as predominant causative agents of candidemia, demonstrating higher virulence factor expression and antifungal resistance compared to C. albicans. The correlation between virulence factors, antifungal resistance, and mortality highlights the importance of species-level identification and antifungal susceptibility testing for optimal management of candidemia.

Systemic fungal infections cause an estimated 3.8 million deaths annually, approximately 10% of which are caused by drug-resistant infections. With only five classes of antifungal drugs, treatment options are limited. Here, we explore synergistic drug combinations-when the efficacy of two drugs combined is greater than expected based on the sum of each individual drug's efficacy-to improve treatment of drug-resistant Cryptococcus neoformans and Candida albicans. Chlorpromazine acts synergistically with both amphotericin B and fluconazole against multiple fungal species, including azole-resistant C. neoformans and C. albicans. We then performed a genome-wide knockout mutant screen and found that ESCRT pathway mutants are resistant to chlorpromazine, while knockout mutants of genes involved in fatty acid biosynthesis are sensitive. Based on these data, we investigated sterol and fatty acid composition in chlorpromazine-treated cells and found only minor increases in sterol precursors, but a substantial increase in lipid droplet size and decreased lipid droplet numbers. This lipid droplet formation potentially sequesters lipid bioavailability and response to membrane stress. Together, these data suggest that chlorpromazine and its analogs are potentially promising treatments for systemic fungal infections that act via lipid homeostasis and stress response.

Disseminated cryptococcosis is a rare but serious opportunistic infection following solid organ transplantation and rarely occurs within the first month of transplantation. We report a case of disseminated Cryptococcus neoformans infection occurring 14 days after orthotopic heart transplantation which required mechanical circulatory support with venoarterial extracorporeal membrane oxygenation (VA-ECMO). Management was complicated by alterations in antifungal pharmacokinetics and the inability to perform serial lumbar punctures while anticoagulated. Antifungal therapy with nonliposomal amphotericin B, flucytosine, and fluconazole was administered. A combination of optic nerve sheath diameter measurements, CT scans, and evaluation for papilledema was used as temporizing, noninvasive surrogates for intracranial pressure monitoring until lumbar punctures could be safely performed. The patient was successfully decannulated, weaned from mechanical support, and discharged from the intensive care unit.

We report a one-step biosynthesis of silver nanoparticles (Fa-AgNPs) using an aqueous Fagonia indica extract and benchmark their activity at the nano-fungal biointerface against Candida albicans and the emerging, multidrug-resistant pathogen Candida auris. Fa-AgNPs were polydisperse and quasi-spherical, with a smaller mean diameter than citrate-capped chemically synthesized AgNPs (Cs-AgNPs) (19.41 ± 7.32 nm vs. 29.01 ± 2.63 nm). Fa-AgNPs outperformed Cs-AgNPs on both species, with MICs of 31.25 vs 125 µg/mL against C. albicans and 3.91 vs 7.81 µg/mL against azole-resistant C. auris. Against C. auris, Fa-AgNPs were at least 64-fold more potent than fluconazole (3.91 vs >250 µg/mL). To our knowledge, this is the first report of F. indica-mediated AgNPs achieving an MIC below 4 µg/mL against this pathogen. Mechanistically, Fa-AgNPs delayed the onset of growth, elevated intracellular ROS in C. albicans, and induced catalase-based antioxidant defense in both species; SEM showed loss of budding cells and extracellular polymeric substances, together with a transition from smooth to rough surfaces and cell-wall damage, in both species, consistent with a primarily oxidative interfacial mechanism. Hemolysis assays indicated acceptable blood compatibility at the lowest concentration tested (2.5 µg/mL), with hemolysis increasing at higher concentrations.

The increasing resistance of Candida albicans to conventional antifungal agents poses a major clinical challenge, underscoring the urgent need for novel therapeutic strategies. Drug combination therapy has emerged as a promising approach to overcome antifungal resistance. In this study, we evaluated the synergistic antifungal activity of dehydronuciferine (DHN), a natural aporphine alkaloid, in combination with fluconazole (FLC) against six clinically drug-resistant C. albicans isolates. Strong synergism was confirmed by checkerboard microdilution assay, with fractional inhibitory concentration index (FICI) values ranging from 0.063 to 0.265. Notably, for strain CA10, the minimum inhibitory concentration of FLC was reduced from >512 to 0.25 μg/mL in the presence of DHN (FICI = 0.125), representing a >2,000-fold reversal of resistance. In a Galleria mellonella infection model, the DHN + FLC combination significantly improved larval survival compared with either monotherapy (P < 0.01). Phenotypic assays demonstrated that the combination markedly suppressed key virulence traits, including hyphal growth, biofilm formation, and epithelial adhesion. Transcriptomic profiling revealed that these phenotypic changes were associated with the downregulation of hypha- and adhesion-related genes and the ergosterol biosynthesis pathway. Together, these findings identify dehydronuciferine as a promising antifungal chemosensitizer that enhances fluconazole efficacy against drug-resistant C. albicans through multitarget perturbation of resistance- and virulence-associated processes. Further mechanistic and preclinical studies are warranted to evaluate its translational potential.IMPORTANCEDrug-resistant Candida albicans infections are increasingly difficult to treat, and the pipeline for new antifungal drugs remains limited. In this study, we show that the natural aporphine alkaloid dehydronuciferine (DHN) acts synergistically with fluconazole to potently inhibit drug-resistant C. albicans by simultaneously suppressing hyphal growth, biofilm formation, and epithelial adhesion. In a Galleria mellonella infection model, the DHN-fluconazole combination significantly improved survival compared with either agent alone. This combination strategy, which pairs an approved antifungal with a natural compound, offers a promising approach to combat antifungal resistance and may accelerate the development of alternative therapeutic options without the need for entirely new drug classes.

Cryptococcal meningitis during pregnancy is rare and associated with maternal and fetal morbidity and mortality. Cryptococcus gattii causes disease in individuals without recognized immunosuppression and is associated with severe neurological manifestations. A 27-year-old apparently immunocompetent woman at 11 weeks of gestation, with occupational exposure in animal science and ecotourism, presented with severe headache and progressive neurological deterioration following mild SARS-CoV-2 infection. Initial cerebrospinal fluid (CSF) analysis showed nonspecific lymphomononuclear meningitis, and treatment for herpetic encephalitis was initiated. After 15 days of deterioration, repeat CSF examination revealed encapsulated yeasts, and culture confirmed C. gattii. Amphotericin B deoxycholate and fluconazole were initiated because first-line therapy was unavailable. Despite intensive care, she developed refractory intracranial hypertension associated with cerebral cryptococcoma, fetal demise at 17 weeks, and brain death. This case highlights diagnostic and therapeutic challenges in cryptococcal central nervous system infection during pregnancy and the importance of considering C. gattii in patients without recognized immunosuppression.

We report a case of refractory coccidioidal meningitis presenting with bilateral upper-extremity weakness in a man-in-a-barrel distribution caused by cervical arachnoiditis, adhesive bands, and spinal cord compression. The patient was treated sequentially with fluconazole, other azole therapy, liposomal amphotericin B, intrathecal amphotericin B, and olorofim, with only partial disease control. Neuroimaging demonstrated progressive cervical leptomeningeal inflammation, cord tethering, and CT myelography illustrated adhesive arachnoiditis. Surgical lysis of adhesions and placement of a spinal Ommaya reservoir for intrathecal amphotericin delivery above and below the obstructed segment resulted in partial neurologic improvement. This case broadens the differential diagnosis of man-in-a-barrel syndrome to include active fungal meningitis with compressive cervical myelopathy and highlights the role of combined surgical-medical management in complicated fungal arachnoiditis.

As part of toxicity assessment in the development of pharmaceuticals and pesticides, ex vivo method using testicular tissue slices is suggested for evaluating the impact and underlying mechanisms of action of compounds on androgen synthesis. However, relevant comprehensive assessments, such as comprehensive profiling of androgens and related hormones and histological examination of tissue slices, remain limited. We evaluated an ex vivo method using testicular tissue slices by determining whether the amounts of each hormone synthesized in successive metabolic steps could be measured to reflect compound-induced effects. First, rat testicular tissue slices were incubated with ketoconazole (Ket) solutions (a representative azole fungicide that inhibits androgen synthesis) for 3 h; hormone assays of the supernatant and histological examination of the slices, including hematoxylin and eosin staining, ultrastructural examination, and immunohistochemistry, were conducted. Changes in androgen and precursor hormone levels were consistent with cytochrome P450 (CYP) 17A1 inhibition by Ket, together with morphological observations in Leydig cells that generally reflected the altered hormone profiles. Subsequently, eight azoles (hexaconazole, flusilazole, triticonazole, imazalil, penconazole, myclobutanil, triadimefon, and fluconazole) with inhibitory effects on CYP17A1 were examined. Hormone assays revealed changes suggestive of CYP17A1 inhibition for all the compounds, along with apparent differences in their inhibitory potency. Histological examination provided supportive morphological observations that were generally consistent with differences in hormone synthesis among the compounds. These results demonstrate that this ex vivo method can detect azole-induced disruptions in androgen synthesis and associated histopathological changes with fewer resources, thus offering a valuable tool for mechanistic toxicity studies.

The lignin extracted from the leaves of Crataeva tapia was investigated for its antimicrobial activity and potential use in solid pharmaceutical formulations. The leaves contained 35% cellulose, 23.4% hemicellulose, 17.3% lignin, 19.1% extractives, and 4.7% ash. The extraction yield after acid precipitation was 67.8%. Cytotoxicity assays in RAW 264.7, J774, Vero, V79, and HepG2 cells showed low toxicity (IC50 >1000 µg/mL) and no hemolytic activity (<10%). Lignin exhibited MIC values ranging from 512 to >1024 µg/mL, with no MBC or MFC detected. The best antibacterial result was observed against Staphylococcus aureus UFPEDA-709 (MIC 512 µg/mL), comparable to oxacillin. Among Candida strains, notable results were obtained for HAM11 (128 µg/mL; fluconazole 0.5 µg/mL) and HAM13 (128 µg/mL; fluconazole 4 µg/mL). No synergism was observed against bacteria, but additive and synergistic effects were detected in HAM11 and HAM13, respectively. Lignin reduced HAM13 biofilm formation by 16%. Ergosterol assays suggested its action on fungal cell walls and membranes. Tablets containing lignin showed hardness of 5.5 Kgf/cm[2], friability of 0.93%, and disintegration in 36.2 min, while those without lignin showed 4.6 Kgf/cm[2], 0.15%, and 31 min, respectively. These findings highlight C. tapia lignin as a versatile and promising biomaterial for sustainable fluconazole-based formulations and adjuvant therapies.

Treatment of symptomatic patients with coccidioidomycosis often involves the use of triazole antifungals. There is concern for reduced susceptibility of Coccidioides to fluconazole, as a high percentage of isolates were previously reported to have elevated fluconazole MICs. We reviewed our recent azole MICs against Coccidioides and examined temporal trends over a 20-year period. Our clinical laboratory database was queried for data against Coccidioides between 2006 and 2025, and trends in GM MICs and the percentage of isolates with elevated MICs to the azoles were assessed. All testing had been performed by broth dilution according to Clinical and Laboratory Standards Institute (CLSI) M38 methods. The results over the 10-year period from 2016 to 2025 were compared to those previously reported by our group from 2001 to 2015. Overall, the results between the two periods showed similar MIC parameters and distributions for fluconazole, itraconazole, posaconazole, and voriconazole, with posaconazole demonstrating the most potent activity, while fluconazole demonstrated reduced susceptibility. Isavuconazole MIC parameters were available during the second period and similar to those of voriconazole. Weak correlations were observed between fluconazole MICs and those of the other azoles. Fluctuations in GM MICs and the percentage of isolates with elevated MICs were observed with fluconazole, itraconazole, and posaconazole, with elevated values observed between 2011 and 2019. Continued surveillance is needed, and outcome data are required to determine if the consistently reduced susceptibility observed with fluconazole or in those infected with strains that have higher MICs results in poorer outcomes in patients with coccidioidomycosis.