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.

• Klíčová slova
• Antifungal, Capsule, Cryptococcus neoformans, Synergism, Verapamil,
• MeSH
• amfotericin B farmakologie   terapeutické užití   MeSH
• antifungální látky farmakologie   terapeutické užití   MeSH
• Cryptococcus neoformans * MeSH
• flucytosin farmakologie   terapeutické užití   MeSH
• kryptokokóza * farmakoterapie   mikrobiologie   MeSH
• mikrobiální testy citlivosti MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• amfotericin B MeSH
• antifungální látky MeSH
• flucytosin 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.

• Klíčová slova
• Antifungal activity, Cryptococcus neoformans, Hydroxychloroquine, Itraconazole, Synergism,
• MeSH
• antifungální látky farmakologie   terapeutické užití   MeSH
• Cryptococcus neoformans * MeSH
• hydroxychlorochin farmakologie   terapeutické užití   MeSH
• itrakonazol farmakologie   MeSH
• kryptokokóza * farmakoterapie   mikrobiologie   MeSH
• mikrobiální testy citlivosti MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• antifungální látky MeSH
• hydroxychlorochin MeSH
• itrakonazol 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.

• Klíčová slova
• Cinnamaldehyde, Cryptococcosis, Giant cells, In vitro, In vivo,
• MeSH
• akrolein analogy a deriváty   farmakologie   MeSH
• analýza přežití MeSH
• antifungální látky farmakologie   MeSH
• Cryptococcus neoformans účinky léků   MeSH
• fungální léková rezistence účinky léků   MeSH
• játra patologie   MeSH
• kryptokokóza farmakoterapie   mikrobiologie   patologie   MeSH
• mikrobiální testy citlivosti MeSH
• modely nemocí na zvířatech MeSH
• mozek patologie   MeSH
• mykózy farmakoterapie   MeSH
• myši MeSH
• plíce patologie   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• akrolein MeSH
• antifungální látky MeSH
• cinnamaldehyde MeSH Prohlížeč

Cryptococcosis is caused by members of the Cryptococcus neoformans/Cryptococcus gattii species complex. Based on molecular identification, these two species have been further differentiated into molecular types. The aim of this work was to characterize clinical cryptococcal isolates recovered from six hospitals in Northeast Mexico from 1995 to 2011. One hundred and sixty-six isolates, which were characterized by biochemical tests and in vitro susceptibility to amphotericin B, fluconazole, and voriconazole, and M13 PCR fingerprinting, were included in this study. Utilizing phenotypic tests, 153 isolates (92.16 %) were identified as C. neoformans and 13 (7.83 %) as C. gattii. All isolates were susceptible to all antifungals tested. Employing M13 PCR fingerprinting, eight molecular types were detected. VNI was the most common genotype (124 cases; 74.6 %), followed by VNII (15 cases; 9 %), VNIII (8 cases; 4.8 %), VNIV (6 cases; 3.6 %), VGI (6 cases; 3.6 %), VGII (3 cases; 1.8 %), and VGIII and VGIV (2 cases, 1.2 % each). We confirm the presence of C. gattii in clinical isolates in Northeast Mexico, and a high clonal diversity in the studied strains of C. neoformans/C. gattii species complex.

• MeSH
• antifungální látky farmakologie   MeSH
• Cryptococcus gattii klasifikace   genetika   izolace a purifikace   MeSH
• Cryptococcus neoformans klasifikace   genetika   izolace a purifikace   MeSH
• DNA fingerprinting MeSH
• dospělí MeSH
• kryptokokóza epidemiologie   mikrobiologie   MeSH
• lidé středního věku MeSH
• lidé MeSH
• mikrobiální testy citlivosti MeSH
• mladiství MeSH
• mladý dospělý MeSH
• molekulární epidemiologie MeSH
• molekulární typizace * MeSH
• mykologické určovací techniky * MeSH
• nemocnice MeSH
• polymerázová řetězová reakce MeSH
• senioři MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• mladý dospělý MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Mexiko epidemiologie   MeSH
• Názvy látek
• antifungální látky MeSH

BACKGROUND: This basic research aimed to investigate the effects of the actin inhibitor latrunculin A (LA) on the human pathogen Cryptococcus neoformans, by freeze-substitution (FS) and electron microscopy (EM), to determine whether the actin cytoskeleton can become a new antifungal target for inhibition of cell division. METHODS: Cells treated with LA for 20 h in yeast-extract peptone dextrose medium were investigated by phase-contrast and fluorescent microscopy, FS and transmission EM, counted in a Bürker chamber and the absorbance was then measured. RESULTS: The disappearance of actin patches, actin cables and actin rings demonstrated the response of the cells of C. neoformans to the presence of the actin inhibitor LA. The removal of actin cables and patches arrested proliferation and led to the production of cells that had ultrastructural disorder, irregular morphology of the mitochondria and thick aberrant cell walls. Budding cells lysed in the buds and septa. CONCLUSION: LA exerts fungistatic, fungicidal and fungilytic effects on the human pathogenic yeast C. neoformans.

• MeSH
• aktiny antagonisté a inhibitory   MeSH
• antifungální látky farmakologie   MeSH
• bicyklické sloučeniny heterocyklické farmakologie   MeSH
• buněčná stěna účinky léků   metabolismus   MeSH
• buněčné dělení účinky léků   MeSH
• Cryptococcus neoformans účinky léků   metabolismus   MeSH
• elektronová mikroskopie metody   MeSH
• fluorescenční mikroskopie metody   MeSH
• kryptokokóza farmakoterapie   metabolismus   mikrobiologie   MeSH
• lidé MeSH
• mikrofilamenta účinky léků   metabolismus   MeSH
• proliferace buněk účinky léků   MeSH
• thiazolidiny farmakologie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• aktiny MeSH
• antifungální látky MeSH
• bicyklické sloučeniny heterocyklické MeSH
• latrunculin A MeSH Prohlížeč
• thiazolidiny MeSH

Growth patterns of Cryptococcus neoformans submerged culture in different culture volumes, intensity of agitation and types of sealing were evaluated to better understand the physiological role of hypoxia response in this yeast. When low intensity agitation was set at high culture volumes and air exchange between the cultivation vessel and external environment was not abolished completely, the cells proliferated slowly but steadily. On the other hand, when the intensity of agitation was high but the vessel was withheld from fresh air supply, the cells first proliferated rapidly, then arrested completely and finally died. Therefore, the central strategy of C. neoformans here seems to lie in its proliferation-rate adjustment to the available oxygen levels and not in its capacity to survive under anoxia. The data support the opinion that the cultures grown under limited aeration (even though not completely withheld from fresh air supply) are much closer to the real cryptococcal life in human tissues than conventional well-aerated exponential cultures.

• MeSH
• Cryptococcus neoformans růst a vývoj   metabolismus   patogenita   MeSH
• kryptokokóza mikrobiologie   MeSH
• kultivační média metabolismus   MeSH
• kultivační techniky metody   MeSH
• kyslík metabolismus   MeSH
• lidé MeSH
• mikrobiální viabilita MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• hodnotící studie MeSH
• práce podpořená grantem MeSH
• Názvy látek
• kultivační média MeSH
• kyslík MeSH

The cryptococcal polysaccharide antigen was detected in 10 cerebrospinal fluid (CSF) and 23 serum samples from cryptococcal meningitis and intestinal cryptococcosis by the cryptococcal antigen latex agglutination system (CALAS). CALAS titers in CSF and serum samples of cryptococcal meningitis ranged over 8-2048 and 32-2048, respectively, while in cases of intestinal cryptococcosis, serum titers ranged over 8-2048. The isolates of yeast Cryptococcus neoformans were determined to be of serotype A or of the A/D pair. The total leukocyte count and biochemical parameters in CSF were significantly increased as indicators of microbial infection. Furthermore, the in vitro change of the teleomorph (sexual state) to the anamorph (asexual state) was also detected and the teleomorph state changed in vivo to the encapsulated anamoph state which is more virulent during infection in vivo than the yeast-like noncapsulated form. Two primers for internal transcribed spacer (ITS) regions of ribosomal DNA were used for molecular detection of C. neoformans. After PCR amplification, a DNA band of 415 bp, visualized on agarose gel, indicated the presence of C. neoformans cells in the tested CSF and serum samples. The primer sensitivity was also characterized using purified yeast chromosomal DNA as template; it was about 20 pg or more chromosomal DNA which represents about 10 cells of C. neoformans. The primers were also specific for ITS regions of C. neoformans and gave negative results with Candida albicans and E. coli chromosomal DNA templates.

• MeSH
• Cryptococcus neoformans klasifikace   genetika   izolace a purifikace   MeSH
• dítě MeSH
• DNA fungální krev   mozkomíšní mok   MeSH
• dospělí MeSH
• kryptokoková meningitida diagnóza   mikrobiologie   MeSH
• kryptokokóza diagnóza   mikrobiologie   MeSH
• latex fixační testy MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• mykologické určovací techniky MeSH
• nemoci střev diagnóza   mikrobiologie   MeSH
• polymerázová řetězová reakce metody   MeSH
• polysacharidy krev   mozkomíšní mok   MeSH
• předškolní dítě MeSH
• senzitivita a specificita MeSH
• sérotypizace MeSH
• Check Tag
• dítě MeSH
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• předškolní dítě MeSH
• Publikační typ
• časopisecké články MeSH
• hodnotící studie MeSH
• Geografické názvy
• Egypt MeSH
• Názvy látek
• cryptococcal polysaccharide MeSH Prohlížeč
• DNA fungální MeSH
• polysacharidy MeSH

The proportion of yeast species involved in eye infections in 11 patients was examined. The presence of yeast organisms as causative agents of endophthalmitis was found in corneal smears (n = 4), conjunctival swabs (4), and vitreous fluid (3). Altogether 5 strains of Candida albicans, 2 strains of C. krusei and one strain each of C. guilliermondii, C. parapsilosis, C. tropicalis and Cryptococcus neoformans were isolated from the clinical material. The hematogenic origin of endophthalmitis was proved in 7 cases on the basis of positive blood samples and in 2 cases by the isolation of yeasts from the tip of an intravenous catheter. Endophthalmitis-supporting risk factors such as indwelling intravenous catheters, prolonged use of broad-spectrum antibiotics and chemotherapy, surgical intervention, diabetes mellitus, and malignancy were observed in the patients.

• MeSH
• Candida klasifikace   izolace a purifikace   MeSH
• Cryptococcus neoformans izolace a purifikace   MeSH
• kandidóza mikrobiologie   MeSH
• kryptokokóza mikrobiologie   MeSH
• kvasinky izolace a purifikace   MeSH
• lidé MeSH
• novorozenec MeSH
• oční infekce mykotické mikrobiologie   MeSH
• Check Tag
• lidé MeSH
• novorozenec MeSH
• Publikační typ
• časopisecké články MeSH

Cryptococcus neoformans isolated from various clinical materials in 14 cases, was identified by (1) cultivation on Sabouraud glucose agar and CHROMagar Candida, (2) microscopic examination of Indian-ink-stained preparations and (3) determination of biochemical properties (assimilation and fermentation of saccharides, assimilation of KNO3, production of urease and phenol monooxygenase). C. neoformans was determined in five specimens from paediatric patients in the intensive care unit and in nine specimens from adult patients, most frequently from liquor at meningitis (n = 3).

• MeSH
• Cryptococcus neoformans izolace a purifikace   MeSH
• dospělí MeSH
• jednotky intenzivní péče MeSH
• kryptokokóza epidemiologie   mikrobiologie   MeSH
• lidé středního věku MeSH
• lidé MeSH
• meningitida mikrobiologie   MeSH
• nemoci nedonošenců mikrobiologie   MeSH
• novorozenec MeSH
• prevalence MeSH
• senioři MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• novorozenec MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Slovenská republika epidemiologie   MeSH

• MeSH
• Cryptococcus izolace a purifikace   MeSH
• kožní nemoci mikrobiologie   MeSH
• kryptokokóza mikrobiologie   MeSH
• kultivační média MeSH
• lidé středního věku MeSH
• lidé MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• kultivační média MeSH