Cryptococcosis is an invasive mycosis caused mainly by Cryptococcus gattii and C. neoformans and is treated with amphotericin B (AMB), fluconazole and 5-fluorocytosine. However, antifungal resistance, limited and toxic antifungal arsenal stimulate the search for therapeutic strategies such as drug repurposing. Among the repurposed drugs studied, the selective serotonin reuptake inhibitors (SSRIs) have shown activity against Cryptococcus spp. However, little is known about the antifungal effect of duloxetine hydrochloride (DH), a selective serotonin and norepinephrine reuptake inhibitor (SSNRI), against C. neoformans and C. gattii. In this study, DH inhibited the growth of several C. neoformans and C. gattii strains at concentrations ranging from 15.62 to 62.50 μg/mL. In addition, DH exhibited fungicidal activity ranging from 15.62 to 250 μg/mL. In biofilm, DH treatment reduced Cryptococcus spp. biomass at a level comparable to AMB, with a significant reduction (85%) for C. neoformans biofilms. The metabolic activity of C. neoformans and C. gattii biofilms decreased significantly (99%) after treatment with DH. Scanning electron micrographs confirmed the anti-biofilm activity of DH, as isolated cells could be observed after treatment. In conclusion, DH showed promising antifungal activity against planktonic cells and biofilms of C. neoformans and C. gattii, opening perspectives for further studies with DH in vivo.

• MeSH
• Antifungal Agents * pharmacology   MeSH
• Biofilms * drug effects   MeSH
• Cryptococcus gattii * drug effects   MeSH
• Cryptococcus neoformans * drug effects   growth & development   MeSH
• Duloxetine Hydrochloride * pharmacology   MeSH
• Cryptococcosis drug therapy   microbiology   MeSH
• Microbial Sensitivity Tests * MeSH
• Publication type
• Journal Article MeSH

Cryptococcus neoformans is an encapsulated yeast that can cause cryptococcosis and cryptococcal meningitis, which conventional treatment involves antifungal drugs such as polyenes, flucytosine, azoles, and their combinations. However, the high cost, toxicity, and increase in fungi resistance to antifungal agents stimulate the search for therapeutic strategies such as drug repurposing and combination therapy. This study evaluated the activity of the antihypertensive verapamil (VEH) alone and combined with amphotericin B (AmB) against C. neoformans. VEH exhibited antifungal activity against C. neoformans with minimum inhibitory concentration and minimum fungicidal concentration of 118 μg per mL. The combination of VEH and AmB exhibited synergism, reducing at least eightfold both drugs' concentrations. Moreover, the combination decreased the size and glucuronoxylomannnan content of C. neoformans capsule. However, no difference was observed in ergosterol levels of C. neoformans after treatment with VEH and AmB in combination. Altogether, VEH in combination with AmB exhibits potential as a candidate as for the development of anti-cryptococcal drug.

• MeSH
• Amphotericin B pharmacology   therapeutic use   MeSH
• Antifungal Agents pharmacology   therapeutic use   MeSH
• Cryptococcus neoformans * MeSH
• Flucytosine pharmacology   therapeutic use   MeSH
• Cryptococcosis * drug therapy   microbiology   MeSH
• Microbial Sensitivity Tests MeSH
• Publication type
• Journal Article MeSH

Cryptococcus neoformans is an opportunistic fungal pathogen that can cause life-threatening invasive fungal infections. As its prevalence and drug resistance continue to rise, cryptococcosis requires new treatment options. Tapping into the potential antifungal effects of traditional drugs or combination therapy has become one of the options. This study was the first to examine the interaction of hydroxychloroquine (HCQ) and itraconazole (ITR) on Cryptococcus neoformans in vitro and in vivo. Our results showed that HCQ alone and in combination with ITR exhibited antifungal activity against C. neoformans planktonic cells. When HCQ was combined with ITR, the minimal inhibitory concentration (MIC) value of HCQ decreased to 32 μg/mL, and the MIC value of ITR decreased from 0.25 μg/mL to 0.06-0.25 μg/mL. The time-killing curve showed that the combined application of HCQ and ITR significantly shortened the killing time, dynamically defining the antifungal activity. The minimum biofilm clearance concentration (MBEC) of HCQ was only 32 μg/mL, which was significantly lower than the MIC of HCQ for planktonic cells. When combined with ITR, the MBEC of ITR decreased from 128 μg/mL to 2-1 μg/mL, and the MBEC of HCQ decreased from 32 μg/mL to 4 μg/mL, indicating a synergistic antifungal biofilm effect. In comparison to ITR alone, the combination of HCQ and ITR treatment increased the survival of C. neoformans-infected Galleria mellonella larvae and decreased the fungal burden of infected larvae. Mechanistic investigations revealed that HCQ might damage C. neoformans cell membranes, impact the structure of fungal cells, cause extracellular material leakage, and have a potent affinity for attaching to the C. neoformans genomic DNA. In conclusion, HCQ has potential clinical application in the treatment of cryptococcosis.

• MeSH
• Antifungal Agents pharmacology   therapeutic use   MeSH
• Cryptococcus neoformans * MeSH
• Hydroxychloroquine pharmacology   therapeutic use   MeSH
• Itraconazole pharmacology   MeSH
• Cryptococcosis * drug therapy   microbiology   MeSH
• Microbial Sensitivity Tests MeSH
• Animals MeSH
• Check Tag
• Animals MeSH
• Publication type
• Journal Article MeSH

Popisujeme prípad 50-ročnej imunokompetentnej ženy, ktorá bola hospitalizovaná s poruchou vedomia. Stav progredoval do bezvedomia s kvadruparézou, pričom ani laboratórne vyšetrenia ani opakované vyšetrenia likvoru z lumbálnej punkcie nezistili pôvodcu klinického stavu. Pacientka zomrela po osemdňovej hospitalizácii. Mikroskopicky a kultivačne bol po pitve ako etiologický agens zistený Cryptococcus, ktorý počas života viedol k ťažkej kryptokokovej meningoencefalitíde, ktorá bola príčinou smrti. Histologicky boli v tkanive mozgu prítomné nakopenia kryptokokov, ktoré vo farbení hematoxylínom-eozínom verne napodobovali erytrocyty a rozsiahle krvácania. Multifokálna obliterácia cievneho riečiska mozgu kvasinkami bola sprevádzaná rozvojom hypoxicko-ischemických zmien imitujúcich úrazové difúzne axonálne poškodenie mozgu.

In this article, we report the autopsy findings of a 50-year-old immunocompetent woman, who was hospitalized with an altered state of consciousness. Examinations, including cerebrospinal fluid analysis, carried out during hospitalization failed to identify the infectious agent causing progressive loss of consciousness and quadriparesis. The patient died within 8 days of admission to the hospital. Post-mortem microscopic and culture examination revealed Cryptococcus species. Death was attributed to cryptococcal meningoencephalitis. Histologic examination revealed accumulation of cryptococcus mimicking erythrocytes and extensive hemorrhage in hematoxylin and eosin-stained sections of the brain. Multifocal obliteration of the vascular bed by yeast was accompanied by hypoxic-ischemic brain injury mimicking traumatic diffuse axonal injury.

• MeSH
• Cryptococcus * MeSH
• Diffuse Axonal Injury * MeSH
• Middle Aged MeSH
• Humans MeSH
• Meningoencephalitis * diagnosis   MeSH
• Autopsy MeSH
• Brain Injuries MeSH
• Treatment Outcome MeSH
• Check Tag
• Middle Aged MeSH
• Humans MeSH
• Female MeSH
• Publication type
• Case Reports MeSH

Cryptococcosis is an invasive infection that accounts for 15% of AIDS-related fatalities. Still, treating cryptococcosis remains a significant challenge due to the poor availability of effective antifungal therapies and emergence of drug resistance. Interestingly, protease inhibitor components of antiretroviral therapy regimens have shown some clinical benefits in these opportunistic infections. We investigated Major aspartyl peptidase 1 (May1), a secreted Cryptococcus neoformans protease, as a possible target for the development of drugs that act against both fungal and retroviral aspartyl proteases. Here, we describe the biochemical characterization of May1, present its high-resolution X-ray structure, and provide its substrate specificity analysis. Through combinatorial screening of 11,520 compounds, we identified a potent inhibitor of May1 and HIV protease. This dual-specificity inhibitor exhibits antifungal activity in yeast culture, low cytotoxicity, and low off-target activity against host proteases and could thus serve as a lead compound for further development of May1 and HIV protease inhibitors.

• MeSH
• Antifungal Agents chemistry   metabolism   pharmacology   MeSH
• Aspartic Acid Proteases antagonists & inhibitors   genetics   metabolism   MeSH
• Cryptococcus neoformans enzymology   MeSH
• Fungal Proteins antagonists & inhibitors   genetics   metabolism   MeSH
• HIV Protease chemistry   metabolism   MeSH
• HIV enzymology   MeSH
• Fungi drug effects   MeSH
• Catalytic Domain MeSH
• Crystallography, X-Ray MeSH
• Drug Evaluation, Preclinical MeSH
• Recombinant Proteins biosynthesis   chemistry   isolation & purification   MeSH
• Molecular Dynamics Simulation MeSH
• Substrate Specificity MeSH
• Binding Sites MeSH
• Structure-Activity Relationship MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Research Support, N.I.H., Extramural MeSH

Cryptococcosis is a potentially fatal fungal disease which has aggrandized with the emergence of AIDS and antifungal resistance. The currently used antifungals lack the broad-spectrum activity and result in several toxicities during long treatment regimens. Thus, the present study aims to evaluate the antifungal activity of cinnamaldehyde against Cryptococcus neoformans var. grubii, the etiological agent of the disease. Quantitative and qualitative in vitro fungal susceptibilities were carried out by minimum inhibitory concentration assay, flow cytometric analysis, and confocal microscopy. Micromorphological alterations were studied through scanning electron and light microscopies. "In vivo" antifungal efficacy of cinnamaldehyde was assessed. Cinnamaldehyde showed antifungal activity against C. neoformans in a dose-dependent manner. A concentration of 1.37 mg/mL of cinnamaldehyde was found to be inhibitory and fungicidal while the low concentration (0.68 mg/mL) was found to induce micromorphological changes and formation of giant/titan-like cells in this pathogen. The reparative activity of cinnamaldehyde and its ability to prolong the life even after the advent of cryptococcal meningitis in mice was also noticed. This study suggests potent anti-cryptococcal activity of cinnamaldehyde. Though, it has a couple of limitations like allergy and low bioavailability. However, these problems can be circumvented by developing suitable analogs of the compound. It, therefore, could be used as a therapeutic option against cryptococcosis and cryptococcal meningitis. Moreover, the evaluation of its pharmacokinetic and pharmacodynamic properties is desirable.

• MeSH
• Acrolein analogs & derivatives   pharmacology   MeSH
• Survival Analysis MeSH
• Antifungal Agents pharmacology   MeSH
• Cryptococcus neoformans drug effects   MeSH
• Drug Resistance, Fungal drug effects   MeSH
• Liver pathology   MeSH
• Cryptococcosis drug therapy   microbiology   pathology   MeSH
• Microbial Sensitivity Tests MeSH
• Disease Models, Animal MeSH
• Brain pathology   MeSH
• Mycoses drug therapy   MeSH
• Mice MeSH
• Lung pathology   MeSH
• Animals MeSH
• Check Tag
• Mice MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH

URA5-RFLP is one of the most widely used genotyping methods relating to Cryptococcus neoformans and C. gattii consensus genotype nomenclature. In order to identify a molecular type, this method uses a visual comparison of digested PCR products of tested and reference strains, therefore any anomaly in RFLP patterns of studied isolates makes recognition difficult or impossible. This report describes a strain of VNIV type showing an atypical URA5-RFLP pattern as well as a group of AD hybrids displaying the same anomaly. The atypical RFLP pattern is the result of a point mutation and emergence of a new restriction site. Emergence of the allele presenting a new banding pattern may lead to misidentification using the URA5-RFLP technique; the results of this study as well as the literature data may suggest the spread of the allele in the environment.

• MeSH
• Cryptococcus neoformans classification   genetics   MeSH
• Genotype MeSH
• Genes, Fungal genetics   MeSH
• Environmental Microbiology MeSH
• Mutation MeSH
• Mycological Typing Techniques MeSH
• Orotate Phosphoribosyltransferase genetics   MeSH
• Polymorphism, Restriction Fragment Length MeSH
• Base Sequence MeSH
• Publication type
• Journal Article MeSH

Kryptokoková meningitída je závažná neuroinfekcia spôsobená kvasinkou Cryptococcus neoformans. Často sa vyskytuje ako oportúnna infekcia, vzácne sa môže prejaviť aj u zdravých ľudí. Najčastejším zdrojom infekcie je vdýchnutie infikovaného vtáčieho trusu. V pľúcach a v priľahlých lymfatických uzlinách môžu kryptokoky perzistovať dlhú dobu. Klinické prejavy pľúcnej infekcie nie sú žiadne, alebo sú len mierne. K manifestácii ochorenia dochádza často až po prieniku kryptokokov do CNS. Prezentovaná kazuistika dokumentuje výskyt kryptokokovej meningitídy u imunokompetentnej pacientky. Bola diagnostikovaná mikroskopickým nálezom kvasiniek z cerebrospinálnej tekutiny. Nález bol potvrdený dôkazom kryptokokovej DNA v likvore a následne kultiváciou. Napriek okamžitému zahájeniu antimykotickej terapie a komplexnej starostlivosti sa stav pacientky skončil úmrtím.

Cryptococcal meningitis is a severe neurological infection caused by the yeast Cryptococcus neoformans. It often occurs as an opportunistic infection; rarely, it may be seen in healthy people as well. The most common source of the infection is inhalation of infected bird droppings. The cryptococci may persist in the lungs and nearby lymph nodes for a long time. There are no or mild clinical manifestations of the pulmonary infection. The disease often manifests only after the cryptococci penetrate into the CNS. The case report documents the development of cryptococcal meningitis in an immunocompetent patient. It was diagnosed by microscopic detection of the yeast in the cerebrospinal fluid. The finding was confirmed by detecting cryptococcal DNA in the cerebrospinal fluid and culture. Despite immediate initiation of antifungal therapy and intensive care, the patient died.

• MeSH
• Cryptococcus neoformans MeSH
• Diagnostic Imaging methods   MeSH
• Flucytosine therapeutic use   MeSH
• Clinical Laboratory Techniques methods   MeSH
• Meningitis, Cryptococcal * diagnosis   therapy   MeSH
• Humans MeSH
• Aged MeSH
• Sepsis MeSH
• Death MeSH
• Treatment Outcome MeSH
• Check Tag
• Humans MeSH
• Aged MeSH
• Female MeSH
• Publication type
• Case Reports MeSH
• Research Support, Non-U.S. Gov't MeSH

• MeSH
• Aspergillus fumigatus pathogenicity   MeSH
• Bacterial Infections diagnosis   therapy   MeSH
• Candida albicans pathogenicity   MeSH
• Cryptococcus gattii pathogenicity   MeSH
• Cryptococcus neoformans pathogenicity   MeSH
• Cytomegalovirus Infections diagnostic imaging   diagnosis   therapy   MeSH
• Herpesviridae Infections diagnosis   drug therapy   MeSH
• Epstein-Barr Virus Infections diagnosis   MeSH
• Infections diagnosis   therapy   MeSH
• Humans MeSH
• Mucorales pathogenicity   MeSH
• Mycoses diagnostic imaging   diagnosis   therapy   MeSH
• Pneumocystis pathogenicity   MeSH
• Strongyloides pathogenicity   MeSH
• Toxoplasma pathogenicity   MeSH
• Lung Transplantation * MeSH
• Check Tag
• Humans MeSH

• MeSH
• Antifungal Agents adverse effects   MeSH
• Cryptococcus neoformans isolation & purification   MeSH
• Zebrafish MeSH
• Dexamethasone adverse effects   MeSH
• Intracranial Hypertension MeSH
• Intracranial Pressure physiology   MeSH
• Meningitis, Cryptococcal * drug therapy   immunology   microbiology   MeSH
• Humans MeSH
• Spinal Puncture * methods   MeSH
• Point-of-Care Systems MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Animals MeSH
• Publication type
• Review MeSH