Cryptococcal meningitis has become the largest cause for the death of infectious diseases in the central nervous system infectious disease worldwide. Most patients with cryptococcal meningitis have AIDS, autoimmune diseases, hematologic malignancies, and some other relevant diseases. It is mainly caused by infection with
Antiphospholipid Syndrome/complications* ; Cryptococcus neoformans ; Female ; HIV Infections ; Humans ; Meningitis, Cryptococcal/complications* ; Middle Aged ; Stroke

Betaine derivatives are considered major ingredients of shampoos and are commonly used as antistatic and viscosity-increasing agents. Several studies have also suggested that betaine derivatives can be used as antimicrobial agents. However, the antifungal activity and mechanism of action of betaine derivatives have not yet been fully understood. In this study, we investigated the antifungal activity of six betaine derivatives against Malassezia restricta, which is the most frequently isolated fungus from the human skin and is implicated in the development of dandruff. We found that, among the six betaine derivatives, lauryl betaine showed the most potent antifungal activity. The mechanism of action of lauryl betaine was studied mainly using another phylogenetically close model fungal organism, Cryptococcus neoformans, because of a lack of available genetic manipulation and functional genomics tools for M. restricta. Our genome-wide reverse genetic screening method using the C. neoformans gene deletion mutant library showed that the mutants with mutations in genes for cell membrane synthesis and integrity, particularly ergosterol synthesis, are highly sensitive to lauryl betaine. Furthermore, transcriptome changes in both C. neoformans and M. restricta cells grown in the presence of lauryl betaine were analyzed and the results indicated that the compound mainly affected cell membrane synthesis, particularly ergosterol synthesis. Overall, our data demonstrated that lauryl betaine influences ergosterol synthesis in C. neoformans and that the compound exerts a similar mechanism of action on M. restricta.
Anti-Infective Agents ; Betaine ; Cell Membrane ; Cryptococcus ; Cryptococcus neoformans ; Dandruff ; Ergosterol ; Fungi ; Gene Deletion ; Genetic Testing ; Genomics ; Humans ; Malassezia ; Methods ; Skin ; Transcriptome

OBJECTIVE: To compare the sensitivity and specificity of real-time fluorescent quanttative PCR(FQ-PCR), blood culture and serum capsular antigen test for the detection of blood flow infection with cryptococcus reoformans, so as to provide the experimental evidence for use of FQ-PCR to detect the blood flow infection with cryptococcus neoformans. METHODS: Sixty male Sprague-Dawley (SD) rats were randomly divided into group A (immune suppression plus infection), group B (immune normal plus infection), group C (immune suppression plus non infection) and group D (normal control). The rats in group A were injected intraperitoneally with cyclophosphamide at D1 of experiment and were injected with suspension of cryptococcus neoformans by tail vein at D4 of experiment; the rats in group B were injected intraperitoneally with saline at D1 and were injected with suspension of cryprococcus neoformans by tail vein at D4; the rats in group C were injected intraperitoneally with cyclophosphamide at D1 and were injected with saline by tail vein at D4; the rats in group D were injected intaperitoneally with saline at D1 and were injected with saline by tail vein at D4.At D 4 after successful extablishment of rat model with infection, the blood samples were collected from ocular veneous plexus at 0, 6, 12, 24, 48 and 72 hours by parity number respectively, then the plasma was extracted, and the blood samples infected at different time were detected by FQ-PCR, and at the same time, the blood culture and serum capsular antigen test were performed. The detected results obtained from above-mentioned 3 kinds of detection methods were compared. RESULTS: The FQ-PCR detection of cryptococcus neoformoms showed that the positive rate detected after 12 hours in A group significantly increased (P<0.05), as compared with B, C and D groups. For the blood samples, the positive rate detected by FQ-PCR was significantly higher than that detected by the blood culture and serum capsular antigen test, moreover the detected results could be quantified, and difference was statistically significant (P<0.05). CONCLUSION The FQ-PCR system for detection of cryptococcus neoformant can detect the pathogens in blood of infected rats, and its sensitivity is superior to the blood culture and serum capsular antigen test; the FQ-PCR can detect the pathogens in blood of infected rats much more early, as compared with the blood culture and serum capsular antigen test.
Animals ; Cryptococcus neoformans ; Female ; Male ; Pregnancy ; Rats ; Rats, Sprague-Dawley ; Real-Time Polymerase Chain Reaction ; Sensitivity and Specificity

Cryptococcus neoformans is the most common microorganism found in cerebrospinal fluid (CSF) cytology and causes life-threatening infections in immunocompromised hosts. Although its cytomorphologic features in conventional smear cytology have been well described, those in liquid-based cytology have rarely been. A 73-year-old woman with diffuse large B-cell lymphoma presented with mental confusion and a spiking fever. To rule out infectious conditions, CSF examination was performed. A cytology slide that was prepared using the ThinPrep method showed numerous spherical yeast-form organisms with diameters of 4–11 μm and thick capsules. Occasional asymmetrical, narrow-based budding but no true hyphae or pseudohyphae were observed. Gomori methenamine silver staining was positive. Cryptococcosis was confirmed in blood and CSF through the cryptococcal antigen test and culture. Liquid-based cytology allows for a clean background and additional slides for ancillary testing, facilitating the detection of microorganisms in CSF specimens, particularly when the number of organisms is small.
Aged ; Capsules ; Cerebrospinal Fluid ; Cryptococcosis ; Cryptococcus neoformans ; Female ; Fever ; Humans ; Hyphae ; Immunocompromised Host ; Lymphoma, B-Cell ; Meningitis, Cryptococcal ; Methenamine ; Methods

Mon1 is a guanine nucleotide exchange factor subunit that activates the Ypt7 Rab GTPase and is essential for vacuole trafficking and autophagy in eukaryotic organisms. Here, we identified and characterized the function of Mon1, an ortholog of Saccharomyces cerevisiae Mon1, in a human fungal pathogen, Cryptococcus neoformans. Mutation in mon1 resulted in hypersensitivity to thermal stress. The mon1 deletion mutant exhibited increased sensitivity to cell wall and endoplasmic reticulum stress. However, the mon1 deletion mutant showed more resistance to the antifungal agent fluconazole. In vivo studies demonstrated that compared to the wild-type strain, the mon1 deletion mutant attenuated virulence in the Galleria mellonella insect model. Moreover, the mon1 deletion mutant was avirulent in the murine inhalation model. These results demonstrate that Mon1 plays a crucial role in stress survival and pathogenicity in C. neoformans.
Autophagy ; Cell Wall ; Cryptococcus neoformans* ; Cryptococcus* ; Endoplasmic Reticulum Stress ; Fluconazole ; GTP Phosphohydrolases ; Guanine Nucleotide Exchange Factors ; Humans ; Hypersensitivity ; Inhalation ; Insects ; Saccharomyces cerevisiae ; Vacuoles ; Virulence*

BACKGROUND: Meningitis is a clinically important disease because of its high mortality and morbidity. The epidemiology of this disease has changed remarkably due to the introduction of pneumococcal vaccines and Haemophilus influenzae type b (Hib) conjugate vaccine. Therefore, it is required to continuously monitor and research the organisms isolated from cerebrospinal fluid (CSF) cultures. METHODS: We analyzed trends of bacteria and fungi isolates obtained from CSF cultures between 1997 and 2016 in a tertiary care hospital according to year, month, gender, and age. RESULTS: Out of a total of 38,450 samples, we identified 504 (1.3%) isolates. The isolation rate in the first tested decade (1997–2006) ranged from 1.3% to 3.1%, while that in the second decade (2007–2016) ranged from 0.4% to 1.5%. The most common organisms was coagulase-negative staphylococci (CoNS) (31.9%), followed by Staphylococcus aureus (9.5%), Streptococcus pneumoniae (7.5%), Acinetobacter baumannii (5.8%), and Mycobacterium tuberculosis (5.8%). Monthly isolation rates were highest in May and July and lowest in February and December. Male to female ratio was 1.5:1. The isolation rates of S. pneumoniae, Enterococcus faecium, and Escherichia coli were similar in children and adults, but those of S. aureus, E. faecalis, A. baumannii, Pseudomonas aeruginosa, M. tuberculosis, and Cryptococcus neoformans were higher in adults than in children. CONCLUSION: During the last two decades, the isolation rate of CSF culture per year has decreased, with monthly isolation rates being highest in May and July. CoNS, S. aureus, and S. pneumoniae were most common in males, whereas CoNS, S. pneumoniae, and M. tuberculosis were most common in females. While Group B Streptococcus was most common in infants younger than 1 year, S. aureus and C. neoformans were more common in adults.
Acinetobacter baumannii ; Adult ; Bacteria* ; Cerebrospinal Fluid* ; Child ; Cryptococcus neoformans ; Enterococcus faecium ; Epidemiology ; Escherichia coli ; Female ; Fungi* ; Haemophilus influenzae type b ; Humans ; Infant ; Male ; Meningitis ; Mortality ; Mycobacterium tuberculosis ; Pneumococcal Vaccines ; Pneumonia ; Pseudomonas aeruginosa ; Staphylococcus aureus ; Streptococcus ; Streptococcus pneumoniae ; Tertiary Healthcare* ; Tuberculosis

BACKGROUND: Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) allows rapid and accurate identification of clinical yeast isolates. In-tube formic acid/acetonitrile (FA/ACN) extraction is recommended prior to the analysis with MALDI Biotyper, but the direct on-plate FA extraction is simpler. We compared the Biotyper with the VITEK MS for the identification of various clinically relevant yeast species, focusing on the use of the FA extraction method. METHODS: We analyzed 309 clinical isolates of 42 yeast species (four common Candida species, Cryptococcus neoformans, and 37 uncommon yeast species) using the Biotyper and VITEK MS systems. FA extraction was used initially for all isolates. If ‘no identification' result was obtained following the initial FA extraction, these samples were then retested by using FA (both systems, additive FA) or FA/ACN (Biotyper only, additive FA/ACN) extraction. These results were compared with those obtained by sequence-based identification. RESULTS: Both systems correctly identified all 158 isolates of the four common Candida species after the initial FA extraction. The Biotyper correctly identified 8.7%, 30.4%, and 100% of 23 C. neoformans isolates after performing initial FA, additive FA, and FA/ACN extractions, respectively, while VITEK MS identified all C. neoformans isolates after the initial FA extraction. Both systems had comparable identification rates of 37 uncommon yeast species (128 isolates), following the initial FA (Biotyper, 74.2%; VITEK MS, 73.4%) or additive FA (Biotyper, 82.0%; VITEK MS, 73.4%). CONCLUSIONS: The identification rate of most common and uncommon yeast isolates is comparable between simple FA extraction/Biotyper method and VITEK MS methods, but FA/ACN extraction is necessary for C. neoformans identification by Biotyper.
Candida ; Cryptococcus neoformans ; Mass Spectrometry* ; Methods* ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; Yeasts*

Cryptococcus spp. other than Cryptococcus neoformans or Cryptococcus gattii were previously considered saprophytes and thought to be non-pathogenic to humans. However, opportunistic infections associated with non-neoformans and non-gattii species, such as Cryptococcus laurentii and Cryptococcus albidus, have increased over the past four decades. We experienced a case of cryptococcosis caused by non-neoformans and non-gattii spp. in a 47-year-old female with refractory acute myeloid leukemia after allogeneic hematopoietic stem cell transplantation. The patient underwent salvage chemotherapy with fluconazole prophylaxis and subsequently developed neutropenic fever with multiple erythematous umbilicated papules. A skin biopsy revealed fungal hyphae and repetitive blood cultures showed yeast microorganisms that were identified later as C. laurentii by Vitek-II®. Skin lesions and fever began to improve with conventional amphotericin B therapy. The treatment regimen was continued for 21 days until the disseminated cryptococcosis was completely controlled.
Amphotericin B ; Biopsy ; Cryptococcosis ; Cryptococcus gattii ; Cryptococcus neoformans ; Cryptococcus* ; Drug Therapy ; Female ; Fever ; Fluconazole ; Hematopoietic Stem Cell Transplantation ; Humans ; Hyphae ; Leukemia, Myeloid, Acute* ; Middle Aged ; Opportunistic Infections ; Skin Manifestations* ; Skin* ; Yeasts

Primary cutaneous cryptococcosis is isolated cutaneous cryptococcosis without evidence of systemic involvement. It usually presents as a solitary nodule in an exposed area, such as head and neck or extremities, and the occurrence of primary cutaneous cryptococcosis in the covered area has been rarely reported in the literature. A healthy 81-year-old immunocompetent woman presented with a solitary reddish scaly plaque on her abdomen without clinical evidence of systemic cryptococcal infection. Histopathologic examination showed numerous encapsulated spores, and the organism was identified as Cryptococcus neoformans in a series of fungal studies. We diagnosed this case as a primary cutaneous cryptococcosis by the findings mentioned above. The patient was successfully treated with surgical excision and oral fluconazole without any side effects.
Abdomen* ; Aged, 80 and over ; Cryptococcosis* ; Cryptococcus neoformans ; Extremities ; Female ; Fluconazole ; Head ; Humans ; Neck ; Spores

Cryptococcus neoformans is a fungus that causes opportunistic infections in immunocompromised hosts. Skin lesions are found in 10-20% of systemic cryptococcal infections, usually secondary to cryptococcemia, while primary cutaneous cryptococcosis with cryptococcemia is very rare. We report a case of rapidly spreading cryptococcal cellulitis in a 64-year-old male on maintenance hemodialysis taking steroids for encapsulated peritoneal sclerosis. Bluish bullous cellulitis developed on the left forearm and spread rapidly to the other forearm. We identified C. neoformans in the blood and skin lesions. We treated him successfully with liposomal amphotericin B and fluconazole for 15 months. We also review the literature.
Amphotericin B ; Cellulitis* ; Cryptococcosis ; Cryptococcus neoformans* ; Fluconazole ; Forearm ; Fungi ; Humans ; Immunocompromised Host ; Male ; Middle Aged ; Opportunistic Infections ; Peritoneal Fibrosis ; Renal Dialysis* ; Skin ; Steroids