Folia Microbiol (Praha), 2003, 48(2), 261 - 8 Characterization of Cryptococcus neoformans var . neoformans serotype A and A/D in samples from Egypt; Abdel-Salam HA; 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. J Chemother, 2003 Apr, 15(2), 124 - 8 Flucytosine and cryptococcosis: which in vitro test is the best predictor of outcome? Viviani MA, Esposto MC, Cogliati M, Tortorano AM. The combination of flucytosine and amphotericin B is first choice treatment for active cryptococcosis . Because of innate or acquired resistance of Cryptococcus neoformans to flucytosine, in vitro testing is mandatory . Yeast nitrogen base (YNB) at pH 7.0 is the recommended medium for the broth microdilution test (NCCLS M27-A) and for the E-test . In order to verify if minimum inhibitory concentrations (MICs) were able to predict treatment outcome, the susceptibility of 24 isolates from 21 patients treated with flucytosine alone or in combination was tested by the broth microdilution, agar dilution and E-test using YNB either at pH 7.0 or at pH 5.4 . Only those MICs obtained on YNB pH 5.4 proved to correlate with treatment outcome . The present study suggests that in vitro susceptibility to flucytosine of C . neoformans isolates should be evaluated on YNB pH 5.4 and the test should be standardized accordingly. Eukaryot Cell, 2003 Jun, 2(3), 422 - 30 Calcineurin is essential for Candida albicans survival in serum and virulence; Blankenship JR et al.; Calcineurin is a calcium-activated protein phosphatase that is the target of the immunosuppressants cyclosporin A and FK506 . In T cells, calcineurin controls nuclear import of the NF-AT transcription factor and gene activation . In plants and fungi, calcineurin functions in stress responses (e.g., temperature, cations, and pH) and is necessary for the virulence of the fungal pathogen Cryptococcus neoformans . Here we show that calcineurin is also required for the virulence of another major fungus that is pathogenic to humans, Candida albicans . C . albicans calcineurin mutants had significantly reduced virulence in a murine model of systemic infection . In contrast to its role in C . neoformans, calcineurin was not required for C . albicans survival at 37 degrees C . Moreover, C . albicans calcineurin mutant strains exhibited no defects in known Candida virulence traits associated with host invasion, including filamentous growth, germ tube formation, and adherence to and injury of mammalian cells . C . albicans calcineurin mutant strains failed to colonize and grow in the kidneys of infected animals and were unable to survive when exposed to serum in vitro . Our studies illustrate that calcineurin has evolved to control aspects of the virulence of two divergent fungal pathogens via distinct mechanisms that can be targeted to achieve broad-spectrum antifungal action. Yeast, 2003 Jun, 20(8), 731 - 8 Verification of a new gene on Saccharomyces cerevisiae chromosome III; Zhang Z et al.; We present verification of the existence of a previously unannotated, intron-containing gene of 134 amino acids (predicted molecular weight approximately 15.5 kDa) located on chromosome III of Saccharomyces cerevisiae . Strains carrying a deletion of this gene, which we have called YCL012C, reveal no apparent phenotype . Orthologues of YCL012C are present in related species S . bayanus, S . paradoxus and Ashbya gossypii . Comparison with other fungal sequences reveals that orthologues of this gene are likely present in Schizosaccharomyces pombe, Neurospora crassa and Cryptococcus neoformans as well, indicating that YCL012C is a widely conserved fungal gene . J Immunol, 2003 Jun 15, 170(12), 6125 - 32 OX40 ligation on activated T cells enhances the control of Cryptococcus neoformans and reduces pulmonary eosinophilia; Humphreys IR et al.; Pulmonary eosinophilia induced in C57BL/6 mice after Cryptococcus neoformans infection is driven by CD4(+) Th2 cells . The immunological mechanisms that protect against eosinophilia are not fully understood . Interaction of OX40 (CD134) and its ligand, OX40L, has been implicated in T cell activation and cell migration . Unlike CD28, OX40 is only expressed on T cells 1-2 days after Ag activation . Manipulation of this pathway would therefore target recently activated T cells, leaving the naive repertoire unaffected . In this study, we show that engagement of OX40 by an OX40L:Ig fusion protein drives IFN-gamma production by CD4(+) T cells and reduces eosinophilia and C . neoformans burden in the lung . Using gene-depleted mice, we show that reduction of eosinophilia and pathogen burden requires IL-12 and/or IFN-gamma . C . neoformans infection itself only partially induces OX40L expression by APCs . Provision of exogenous OX40L reveals a critical role of this pathway in the prevention of C . neoformans-induced eosinophilia. Transplantation, 2003 May 15, 75(9), 1544 - 51 Epidemiology of systemic mycoses among renal-transplant recipients in India; Tharayil John G et al.; BACKGROUND: Systemic mycoses have a high impact on tropical renal-transplant recipients . METHODS: Data from 1,476 primary renal-transplant recipients was prospectively recorded from 1986 to 2000 at a single center . Cumulative incidence of systemic mycoses, its time of occurrence, risk factors, outcome, and postmortem findings in 30 patients with systemic mycoses were analyzed . RESULTS: A total of 110 episodes of systemic mycoses occurred in 98 patients . The fungal genera Aspergillus, Cryptococcus, and Candida constituted 61% of pathogens, 45% localizing to the lungs . Cytomegalovirus (CMV) disease caused a 5-fold and chronic liver disease a 2-fold increase in systemic mycoses . Tuberculosis (TB) with or without nocardiosis was a significant coinfection . Cyclosporine (CsA) was associated with nearly a 4-fold risk of systemic mycoses less than 6 months from the time of transplantation as compared with prednisolone+azathioprine (PRED+AZA) therapy . Overall, the probability of survival with systemic mycoses was 73.4%, 60.8%, 39.5%, and 25.6% and was 92.5%, 87.5%, 80.0%, and 75.5% without systemic mycoses at 1, 2, 5, and 10 years, respectively (P<0.0001) . An extended Cox model with time-independent and dependent covariates showed greater than 15 times the risk of death among those who develop systemic mycoses . Similarly, Posttransplantation (postTX) TB+/-Nocardiosis, preTX TB, CMV disease, diabetes mellitus, PTDM, chronic liver disease (>40 months), and Pred+AZA immunosuppression (>2 years) had 3.5, 1.5, 2.9, 1.9, 1.4, 1.6, 2.3 times the risk for death, respectively, as compared with those who did not have those risk factors . CONCLUSIONS: There is a recent predominance of Aspergillus among the transplant recipients . The risk factors for systemic mycoses are CMV disease, chronic liver disease, and hyperglycemia, and TB is an important coinfection . Systemic mycoses increased in the early postTX period with CsA . The risk factors for death are systemic mycoses, CMV disease, chronic liver disease (>40 months), diabetes mellitus, and Pred+AZA immunosuppression (>2 years) . Overall, the probability of survival with systemic mycoses was poor; however, survival has recently improved. Rev Med Chil, 2003 Mar, 131(3), 299 - 302 {Antifungal susceptibility of yeasts by Etest . Comparison of 3 media}; Tapia C et al.; BACKGROUND: The increasing frequency of systemic fungal infections and the emergency of secondary resistance to antifungals in the lasts years, has stimulated the use of methods for antifungal susceptibility testing . Etest is an easily performed quantitative method that has a good agreement with the broth microdilution reference method (NCCLS), if appropriate media are used . AIM: To compare the susceptibility to Amphotericin B (AmB) and Fluconazole (Flu) of 22 opportunistic yeast isolates (C albicans (7), C tropicalis (9), C parapsilosis (3) and Cryptococcus neoformans (3) by Etest, using three different media and to choose the best medium for each tested drug . The studied media were RPMI 1640, Casitone (Cas) and Sabouraud . RESULTS: The interpretation of minimal inhibitory concentration (MIC) endpoints on Sabouraud was difficult for AmB . The American Type Culture Collection (ATCC) strains MICs were out of the acceptable range in this medium . RPMI and Cas were suitable media to test AmB and Flu, but best endpoints were obtained for AmB in RPMI and Flu in Cas . CONCLUSIONS: The use of appropriate media for each antifungal drug optimizes the MIC readings by Etest . AmB should be tested in RMPI and Flu in Cas . Sabouraud must not be used. Microbes Infect, 2003 Jun, 5(7), 667 - 75 The origin and maintenance of virulence for the human pathogenic fungus Cryptococcus neoformans; Steenbergen JN et al.; The origin of virulence in environmental fungi that have no requirement for animal hosts in their life cycle is enigmatic . Cryptococcus neoformans is a human pathogenic fungus with virulence factors for mammalian pathogenesis that also contribute to environmental survival . C . neoformans virulence may originate from selection pressures imposed by environmental predators. Mol Microbiol, 2003 Jun, 48(5), 1377 - 87 The Cryptococcus neoformans MAP kinase Mpk1 regulates cell integrity in response to antifungal drugs and loss of calcineurin function; Kraus PR et al.; Cell wall integrity is crucial for fungal growth, development and stress survival . In the model yeast Saccharomyces cerevisiae, the cell integrity Mpk1/Slt2 MAP kinase and calcineurin pathways monitor cell wall integrity and promote cell wall remodelling under stress conditions . We have identified the Cryptococcus neoformans homologue of the S . cerevisiae Mpk1/Slt2 MAP kinase and have characterized its role in the maintenance of cell integrity in response to elevated growth temperature and in the presence of cell wall synthesis inhibitors . C . neoformans Mpk1 is required for growth at 37 degrees C in vitro, and this growth defect is suppressed by osmotic stabilization . C . neoformans mutants lacking Mpk1 are attenuated for virulence in the mouse model of cryptococcosis . Phosphorylation of Mpk1 is induced in response to perturbations of cell wall biosynthesis by the antifungal drugs nikkomycin Z (a chitin synthase inhibitor), caspofungin (a beta-1,3-glucan synthase inhibitor), or FK506 (a calcineurin inhibitor), and mutants lacking Mpk1 display enhanced sensitivity to nikkomycin Z and caspofungin . Lastly, we show that calcineurin and Mpk1 play complementing roles in regulating cell integrity in C . neoformans . Our studies demonstrate that pharmacological inhibition of the cell integrity pathway would enhance the activity of antifungal drugs that target the cell wall. Antonie Van Leeuwenhoek, 2003, 83(2), 167 - 74 Cryptococcus haglerorum, sp . nov., an anamorphic basidiomycetous yeast isolated from nests of the leaf-cutting ant Atta sexdens; Middelhoven WJ et al.; A yeast strain (CBS 8902) was isolated from the nest of a leaf-cutting ant and was shown to be related to Cryptococcus humicola . Sequencing of the D1/D2 region of the 26S ribosomal DNA and physiological characterization revealed a separate taxonomic position . A novel species named Cryptococcus haglerorum is proposed to accommodate strain CBS 8902 that assimilates n-hexadecane and several benzene compounds . Physiological characteristics distinguishing the novel species from some other members of the C . humicola complex are presented . The phylogenetic relationship of these strains to species of the genus Trichosporon Behrend is discussed. Indian J Pathol Microbiol, 2002 Jul, 45(3), 349 - 51 Cryptococcal lymphadenitis and meningitis in human immunodeficiency virus infection--a case report; Das BP et al.; Cryptococcal meningitis (CM) is the commonest life threatening opportunistic fungal disease in Human Immunodeficiency Virus (HIV) infected individuals . But there are very little reports of lymphadenopathy along with cryptococcal meningitis, although cases of pulmonary, Intestinal, Bone marrow and retinal involvement have been described earlier . Here we report a case of cryptococcal meningitis associated with generalized lymphadenopathy and hepatosplenomegaly. Eur J Immunol, 2003 Jun, 33(6), 1744 - 53 Neutropenia alters lung cytokine production in mice and reduces their susceptibility to pulmonary cryptococcosis; Mednick AJ et al.; Neutrophils are generally considered to contribute to host defense through their potent microbicidal activity . However, there is accumulating evidence that neutrophils also have an important regulatory role in establishing the balance of Th1 and Th2 responses . This study investigated the role of neutrophils in defense against pulmonary Cryptococcus neoformans infection using neutrophil-depleted BALB/c mice generated by administering mAb RB6-8C5 . Neutropenic mice with pulmonary infection survived significantly longer than control mice, but there was no difference between groups infected intravenously . On day 1 of infection, neutropenic mice had significantly smaller fungal burdens than control mice . On day 7, neutropenic mice had significantly higher lung concentrations of IL-10, TNF-alpha, IL-4, and IL-12 than control mice, but there was no difference in IFN-gamma and MCP-1 levels . Neutrophils influenced the outcome of cryptococcal infection in mice through mechanisms that did not involve a reduction in early fungal burden . The absence of neutrophils in lung tissue during the initial stages of infection appeared to alter the inflammatory response in a manner that was subsequently beneficial to the host . Higher levels of Th1- and Th2-associated cytokines in neutropenic mice could have simultaneously promoted a strong cellular response while reducing inflammatory damage to the lung . Our results support the emerging concept that neutrophils play an important function in modulating the development of the immune response. Antonie Van Leeuwenhoek, 2003, 83(3), 231 - 3 Cryptococcus antarcticus var . circumpolaris var . nov., a basidiomycetous yeast from Antarctica; Vishniac HS et al.; Cryptococcus antarcticus Vishniac & Kurtzman var . circumpolaris Vishniac and Onofri var . nov . (Filobasidiales, Tremellomycetidae, Hymenomycetes), an anamorphic yeast with ca . 73% nDNA relatedness to Cryptococcus antarcticus var . antarcticus, differs in failure to assimilate raffinose, a lower maximum temperature for growth, fatty acid profile, and in a single nucleotide change in the D2 region of LSU rDNA. J Dermatol, 2003 May, 30(5), 405 - 10 Disseminated cellulitic cryptococcosis in the setting of prednisone monotherapy for pemphigus vulgaris; Yoo SS et al.; Cryptococcal infections are seen mainly in immunocompromised hosts . The cutaneous manifestations of infection can include papules, plaques, nodules, vesicles, ulcers, ecchymosis, and, rarely, cellulitis . Cryptococcal meningitis is a life-threatening complication of this disease . Pemphigus vulgaris is a rare auto-immune blistering disease that can also be life-threatening . Treatment of pemphigus commonly entails both corticosteroids and steroid-sparing agents . We present a case of disseminated cryptococcal infection in a patient with pemphigus vulgaris treated with high dose corticosteroids as monotherapy . This case provides an opportunity to discuss the difficulty of managing two potentially mortal conditions in which the treatment of pemphigus vulgaris may exacerbate the disseminated cryptococcal infection. Extremophiles, 2003 Jun, 7(3), 185 - 93 Epub 2003 Feb 11. Cold-adapted yeasts as producers of cold-active polygalacturonases; Birgisson H et al.; Eight cold-adapted, polygalacturonase-producing yeasts belonging to four species were isolated from frozen environmental samples in Iceland . They were identified as Cystofilobasidium lari-marini, Cystofilobasidium capitatum, Cryptococcus macerans and Cryptococcus aquaticus species by sequence analysis of rDNA regions . Growth behavior of the isolates was investigated . All strains could grow at 2 degrees C . Addition of glucose to pectin-containing culture medium had a repressive effect on enzyme production except for C . aquaticus, which showed increased polygalacturonase activity . Optimal temperature for enzyme production for the Cystofilobasidium strains was 14 degrees C, while that for the Cryptococcus strains was lower . Among the isolates, C . lari-marini S3B produced highest levels of enzyme activity at pH 3.2 . Preliminary characterization of the polygalacturonases in the culture supernatant showed the enzyme from Cystofilobasidium strains to be optimally active at 40 degrees C and pH 5, and that from the Cryptococcus strains at 50 degrees C and pH 4 . The polygalacturonase from C . macerans started to lose activity after 1 h of incubation at 40 degrees C, while that from the other strains had already lost activity at 30 degrees C . All the strains except C . aquaticus produced isoenzymes of polyglacturonase . In addition to polygalacturonase, the Cystofilobasidium strains produced pectin lyase, C . aquaticus pectin esterase, and C . macerans pectin lyase, pectate lyase and pectin esterase. J Occup Environ Med, 2003 May, 45(5), 470 - 8 Adverse human health effects associated with molds in the indoor environment; Hardin BD et al.; Molds are common and important allergens . About 5% of individuals are predicted to have some allergic airway symptoms from molds over their lifetime . However, it should be remembered that molds are not dominant allergens and that the outdoor molds, rather than indoor ones, are the most important . For almost all allergic individuals, the reactions will be limited to rhinitis or asthma; sinusitis may occur secondarily due to obstruction . Rarely do sensitized individuals develop uncommon conditions such as ABPA or AFS . To reduce the risk of developing or exacerbating allergies, mold should not be allowed to grow unchecked indoors . When mold colonization is discovered in the home, school, or office, it should be remediated after the source of the moisture that supports its growth is identified and eliminated . Authoritative guidelines for mold remediation are available . Fungi are rarely significant pathogens for humans . Superficial fungal infections of the skin and nails are relatively common in normal individuals, but those infections are readily treated and generally resolve without complication . Fungal infections of deeper tissues are rare and in general are limited to persons with severely impaired immune systems . The leading pathogenic fungi for persons with nonimpaired immune function, Blastomyces, Coccidioides, Cryptococcus, and Histoplasma, may find their way indoors with outdoor air but normally do not grow or propagate indoors . Due to the ubiquity of fungi in the environment, it is not possible to prevent immunecompromised individuals from being exposed to molds and fungi outside the confines of hospital isolation units . Some molds that propagate indoors may under some conditions produce mycotoxins that can adversely affect living cells and organisms by a variety of mechanisms . Adverse effects of molds and mycotoxins have been recognized for centuries following ingestion of contaminated foods . Occupational diseases are also recognized in association with inhalation exposure to fungi, bacteria, and other organic matter, usually in industrial or agricultural settings . Molds growing indoors are believed by some to cause building-related symptoms . Despite a voluminous literature on the subject, the causal association remains weak and unproven, particularly with respect to causation by mycotoxins . One mold in particular, Stachybotrys chartarum, is blamed for a diverse array of maladies when it is found indoors . Despite its well-known ability to produce mycotoxins under appropriate growth conditions, years of intensive study have failed to establish exposure to S . chartarum in home, school, or office environments as a cause of adverse human health effects . Levels of exposure in the indoor environment, dose-response data in animals, and dose-rate considerations suggest that delivery by the inhalation route of a toxic dose of mycotoxins in the indoor environment is highly unlikely at best, even for the hypothetically most vulnerable subpopulations . Mold spores are present in all indoor environments and cannot be eliminated from them . Normal building materials and furnishings provide ample nutrition for many species of molds, but they can grow and amplify indoors only when there is an adequate supply of moisture . Where mold grows indoors there is an inappropriate source of water that must be corrected before remediation of the mold colonization can succeed . Mold growth in the home, school, or office environment should not be tolerated because mold physically destroys the building materials on which it grows, mold growth is unsightly and may produce offensive odors, and mold is likely to sensitize and produce allergic responses in allergic individuals . Except for persons with severely impaired immune systems, indoor mold is not a source of fungal infections . Current scientific evidence does not support the proposition that human health has been adversely affected by inhaled mycotoxins in home, school, or office environments. J Biochem (Tokyo), 2003 Apr, 133(4), 553 - 62 Xylose reductase from the Basidiomycete fungus Cryptococcus flavus: purification, steady-state kinetic characterization, and detailed analysis of the substrate binding pocket using structure-activity relationships; Mayr P et al.; Xylose reductase has been purified to apparent homogeneity from cell extracts of the fungus Cryptococcus flavus grown on D-xylose as carbon source . The enzyme, the first of its kind from the phylum Basidiomycota, is a functional dimer composed of identical subunits of 35.3 kDa mass and requires NADP(H) for activity . Steady-state kinetic parameters for the reaction, D-xylose + NADPH + H(+)<--> xylitol + NADP(+), have been obtained at pH 7.0 and 25 degrees C . The catalytic efficiency for reduction of D-xylose is 150 times that for oxidation of xylitol . This and the 3-fold tighter binding of NADPH than NADP(+) indicate that the enzyme is primed for unidirectional metabolic function in microbial physiology . Kinetic analysis of enzymic reduction of aldehyde substrates differing in hydrophobic and hydrogen bonding capabilities with binary enzyme-NADPH complex has been used to characterize the substrate-binding pocket of xylose reductase . Total transition state stabilization energy derived from bonding with non-reacting sugar hydroxyls is approximately 15 kJ/mol, with a major contribution of 5-8 kJ/mol made by interactions with the C-2(R) hydroxy group . The aldehyde binding site is approximately 1.2 times more hydrophobic than n-octanol and can accommodate linear alkyl chains of <or=6 carbons . Hydrophobic interactions provide a total binding energy of approximately 10 kJ/mol . Specificity for the aldehyde substrate is achieved through large decreases in apparent K(m) ( approximately 100-fold) and smaller but significant increases in turnover number ( approximately 5-fold) . We observed up to 250-fold preference of xylose reductase for reaction with pyridine carbaldehydes, 4-nitro-benzaldehyde, and alpha-oxo-aldehydes over reaction with D-xylose, perhaps reflecting a secondary role of this enzyme in detoxication metabolism of reactive endogenous aldehydes and compounds of xenobiotic origin. Infect Immun, 2003 Jun, 71(6), 3384 - 91 Prior exposure to live Mycobacterium bovis BCG decreases Cryptococcus neoformans-induced lung eosinophilia in a gamma interferon-dependent manner; Walzl G et al.; Some common childhood infections appear to prevent the development of atopy and asthma . In some Mycobacterium bovis BCG-vaccinated populations, strong delayed-type hypersensitivity responses to mycobacterial antigens are associated with a reduced risk of atopy . Although BCG exposure decreases allergen-induced lung eosinophilia in animal models, little attention has been given to the effect of immunity to BCG on responses against live pathogens . We used the murine Cryptococcus neoformans infection model to investigate whether prior BCG infection can alter such responses . The present study shows that persistent pulmonary BCG infection of C57BL/6 mice induced an increase in gamma interferon, a reduction in interleukin-5, and a decrease in lung eosinophilia during subsequent Cryptococcus infection . This effect was long lasting, depended on the presence of live bacteria, and required persistence of mycobacterial infection in the lung . Reduction of eosinophilia was less prominent after infection with a mutant BCG strain (DeltahspR), which was rapidly cleared from the lungs . These observations have important implications for the development of vaccines designed to prevent Th2-mediated disease and indicate that prior lung BCG vaccination can alter the pattern of subsequent host inflammation. Transpl Int, 2003 May, 16(5), 336 - 40 Epub 2003 Mar 04. Longitudinal study of cryptococcosis in adult solid-organ transplant recipients; Vilchez R et al.; While studies in kidney recipients have found meningitis to be the most common clinical manifestation of cryptococcosis (Cry), it is unclear if the clinical presentation of Cry differs among various solid-organ transplant (SOT) recipients and whether the serum cryptococcal antigen (SCA) might predict the site of infection . We report the clinical manifestations and the correlation with a positive SCA among 55 consecutive SOT recipients diagnosed with Cry at the University of Pittsburgh Medical Center . These included: heart (n=13), lung (n=4), liver (n=28), kidney (n=9) and small bowel (n=1) recipients . While there were no significant differences in the manifestations of Cry in heart and lung recipients, kidney recipients had disseminated disease as the most common presentation (P=0.02) . In contrast, pneumonia (P=0.003) and meningitis (P=0.02) were more frequent than disseminated disease in liver recipients . Positive SCA was higher in patients with disseminated disease and meningitis than in patients with isolated pneumonia (P=0.0001) . Significant differences in the manifestations of Cry were observed among types of SOT populations . A positive SCA may be predictive of dissemination and meningitis, but it may not be sensitive for pulmonary disease. Southeast Asian J Trop Med Public Health, 2002 Dec, 33(4), 822 - 31 The spectrum of HIV-related disease in rural Central Thailand; Inverarity D et al.; To determine the spectrum of HIV-related illnesses presenting to a rural primary and secondary healthcare facility in Central Thailand, a cross-sectional study was conducted . Routinely collected data were extracted from outpatient medical notes for all adult HIV-infected new attenders of the Manorom Christian Hospital Infectious Disease Clinic . Data concerning inpatient admissions of HIV-infected individuals were collected from ward admission books and discharge summaries . Complete data were available for 229 outpatients, 70% of whom were men . The median age at presentation was 31 years for men and 30 years for women . The majority of the outpatients were married (69%) and infected heterosexually (91%) . The commonest conditions requiring admission were cryptococcal meningitis (15%), bacterial pneumonia (12%), extrapulmonary tuberculosis (12%), Pneumocystis carinii pneumonia (7%), cerebral toxoplasmosis (4%) and pulmonary tuberculosis (3%) . Of the patients presenting for the first time, 32% had AIDS-defining illnesses . Presentations with some conditions, such as tuberculosis and septicemia, were fewer than expected . The common opportunistic infections among HIV-infected adults in this rural region are treatable and preventable . Most patients present with advanced disease and earlier diagnosis, through improved access to voluntary counseling and testing, would enable them to receive appropriate preventive therapies and antiretrovirals as they becomes available . The high prevalence of cryptococcal disease suggests that prophylactic anti-fungal therapy may be of value in this area . Septicemia and tuberculosis may be under-diagnosed, highlighting the need for improved diagnostic laboratory facilities or treatment based upon validated clinical algorithms . Community care and palliative care services for HIV-infected individuals will increasingly be required in this region. J Med Assoc Thai, 2003 Apr, 86(4), 293 - 8 The efficacy of fluconazole 600 mg/day versus itraconazole 600 mg/day as consolidation therapy of cryptococcal meningitis in AIDS patients; Mootsikapun P et al.; Cryptococcal meningitis is one of the major complications affecting the central nervous system of patients suffering from AIDS . The results of treatment, when following current recommendation are still unsatisfactory . OBJECTIVE: This study aimed to evaluate the efficacy of a higher than recommended dose of oral fluconazole and itraconazole as consolidation therapy for cryptococcal meningitis in AIDS patients . DESIGN AND METHOD: HIV infected patients with primary cryptococcal meningitis, who had been treated initially with amphotericin B for 2 weeks were included in this study . They were randomized into two groups, to receive either fluconazole 600 mg daily or itraconazole 600 mg daily for 10 weeks . The response towards the two different treatments was clinically defined to be successful, if after 10 weeks of treatment no clinical symptoms and signs of meningitis remained and the cerebrospinal fluid (CSF) fungal culture was negative . RESULTS: The trial was performed from April 1999 to April 2000 at Srinagarind Hospital, Khon Kaen, Thailand . At the beginning of the trial 44 cases were selected, but only 35 patients proved to be suitable for the final evaluation of the study . Out of those, 19 cases were assigned to the fluconazole and 16 cases to the itraconazole group . Ten weeks after treatment, all patients clinically recovered completely . The CSF sterilization rate for the fluconazole group and for the itraconazole group were 100 and 94 per cent respectively . The Fisher's exact test showed no significant difference in the CSF sterilization rate between both groups (p = 0.26) . CONCLUSION: The result of this study indicates that treatment with either 600 mg per day of fluconazole or itraconazole as consolidation treatment have the same efficacy for AIDS patients suffering from cryptococcal meningitis . The results of this study also suggest, comparing the result of this trial with the results of similar trials published somewhere else, that treatment with the higher doses may be superior to treatment regimens using lower doses, as can be judged from the clinical outcome and the results of the mycological cultures. Antonie Van Leeuwenhoek, 2003, 83(1), 57 - 61 Effects of lycorine on growth and effects of L-galactonic acid-gamma-lactone on ascorbic acid biosynthesis in strains of Cryptococcus laurentii isolated from Narcissus pseudonarcissus roots and bulbs; Onofri S et al.; The alkaloid lycorine, which is considered to inhibit the last step in ascorbic acid biosynthesis, is produced by Narcissus pseudonarcissus . The growth of two strains (C1 and C3) of Cryptococcus laurentii isolated from root tips of N . pseudonarcissus is inhibited by lycorine, as is the in vivo production of ascorbic acid from L-galactonic acid-gamma-lactone . In contrast, C . laurentii strain C4, isolated from the lycorine-containing bracts of the bulb, was not inhibited by lycorine and did not contain ascorbic acid when cultivated with or without L-galactonic acid-gamma-lactone. Rev Inst Med Trop Sao Paulo, 2003 Mar-Apr, 45(2), 69 - 74 Epub 2003 May 14. Opportunistic infections in patients with AIDS admitted to an university hospital of the Southeast of Brazil; Nobre V et al.; Opportunistic diseases in HIV-infected patients have changed since the introduction of highly active anti-retroviral therapy (HAART) . This study aims at evaluating the frequency of associated diseases in patients with AIDS admitted to an university hospital of Brazil, before and after HAART . The medical records of 342 HIV-infected patients were reviewed and divided into two groups: group 1 comprised 247 patients before HAART and, group 2, 95 patients after HAART . The male-to-female rate dropped from 5:1 to 2:1for HIV infection . There was an increase in the prevalence of tuberculosis and toxoplasmosis, with a decrease in Kaposi's sarcoma, histoplasmosis and cryptococcosis . A reduction of in-hospital mortality (42.0% vs . 16.9%; p = 0.00002) has also occurred . An agreement between the main clinical diagnoses and autopsy findings was observed in 10 out of 20 cases (50%) . Two patients with disseminated schistosomiasis and 2 with paracoccidioidomycosis are reported . Overall, except for cerebral toxoplasmosis, it has been noticed a smaller proportion of opportunistic conditions related to severe immunosuppression in the post HAART group . There was also a significant reduction in the in-hospital mortality, possibly reflecting improvement in the treatment of the HIV infection. Scand J Infect Dis, 2003, 35(3), 211 - 2 Chronic diarrhoea caused by Cryptococcus neoformans in a non-human immunodeficiency virus-infected patient; Sungkanuparph S et al.; A 43-y-old woman with underlying cervix carcinoma stage IIIB, status postirradiation, and diabetes mellitus with tripathy suffered from chronic diarrhoea for more than 6 months . Stool examination showed few white blood cells and red blood cells . Rectal swab cultures disclosed Cryptococcus neoformans in 2 samples obtained 2 weeks apart . The diarrhoea responded to treatment with oral fluconazole for 4 weeks . Repeated rectal swabs revealed no yeast growth . During follow-up for 4 y, she had no problems with diarrhoea . Chronic diarrhoea caused by C . neoformans is rare and clinical suspicion is needed. Mycopathologia, 2002, 156(3), 171 - 6 Effect of different K+ concentrations on Cryptococcus neoformans phenoloxidase activity; Vidotto V et al.; Melanin synthesis in Cryptococcus neoformans, catalyzed by phenoloxidase activity, is one of the oldest virulence factors known . However, until now, the relationship between melanin production in C . neoformans and its virulence has been poorly understood . Among different chemical compounds only Fe3+ and Cu2+ cations enhance the phenoloxidase activity in C . neoformans . A few reports in the literature describe the influence of different cations on C . neoformans phenoloxidase activity, excluding iron . In this study, 13 C . neoformans strains isolated from AIDS patients and 7 from bird droppings (B.D.), were examined in order to clarify the effect of different K+ concentrations on phenoloxidase activity . A new solid and liquid caffeic acid minimal synthetic medium (MSM-CAF) containing only caffeic acid and ferric citrate with different potassium concentrations was used to evaluate C . neoformans phenoloxidase activity . In the MSM-CAF solid medium the degree of brown pigmentation on the agar plates was read on days 1, 2 and 3 of incubation, and the pigmentation of the C . neoformans strains was classed into 5 categories . The brown pigment of the liquid MSM-CAF test tubes were checked after 24 hours of incubation by measuring the optical density (O.D.) at 480 nm . Three C . neoformans AIDS and B.D . strains, randomly chosen, were tested for phenoloxidase activity, according to the modified protocols of Polacheck et al., Torres-Guerrero et al . and Rhodes . According to the results obtained, it has been observed that K+ does not activate the phenoloxidase activity in the C . neoformans AIDS and B.D . strains . In particular, with an increase in potassium concentrations in the MSM-CAF solid and liquid medium, there was a corresponding inhibition of the phenoloxidase activity on both the C . neoformans AIDS and B.D . strains. Nippon Ishinkin Gakkai Zasshi, 2003, 44(2), 139 - 44 Immunomagnetic isolation of Cryptococcus neoformans by beads coated with anti-Cryptococcus serum; Katsu M et al.; Immunomagnetic separation (IMS) was utilized for the selective isolation of Cryptococcus neoformans from environmental sources, such as soils and pigeon droppings . Magnetic beads coated with anti-cryptococcal IgG (serotypes A and B) were used to isolate the fungus . In a modeled spiking experiment using C . neoformans serotype A strain and anti-serotype A antibody, the recovery rate of the cells was more than 47% . Specificity experiments using C . neoformans and Candida albicans showed that the beads, when coated with specific antibody for C . neoformans, were highly effective for the separation of C . neoformans strains from C . albicans (more than 97%) . The IMS of serotype B cells with purified anti-serotype B antibody indicated a high specificity . When this IMS technique was applied to soils and pigeon droppings, C . neoformans cells were selectively isolated from 3 out of 8 samples, and C . neoformans DNAs were identified by PCR . Therefore C . neoformans cells were thus selectively isolated and the efficiency of the technique further confirmed. Nippon Ishinkin Gakkai Zasshi, 2003, 44(2), 61 - 4 {Molecular and quantitative analyses of Malassezia microflora on the skin of atopic dermatitis patients and genotyping of M . globosa DNA}; Sugita T et al.; To elucidate the role of Malassezia species in atopic dermatitis (AD) requires investigation of the Malassezia microflora on the skin of AD patients . Previously, M . furfur was considered the dominant species in the microflora, however, this microorganism has been reclassified into five species and reanalysis of the microflora based on the current Malassezia taxonomy is therefore needed . Malassezia is more difficult to isolate and culture than other pathogenic yeasts such as Candida and Cryptococcus, making it difficult to elucidate the microflora of AD patients accurately . We developed a PCR-based non-culture method that does not require the use of isolation or culture techniques . Of the members of the genus Malassezia, M . globosa colonized the skin of both AD patients and healthy subjects more frequently than other Malassezia species . In addition, we found polymorphisms in the intergenic spacer 1 region of the M . globosar RNA gene . The genotypes of the microorganisms obtained from AD patients were significantly different from those obtained from healthy subjects . We believe that a specific genotype of M . globosa is responsible for exacerbation of AD. FEMS Yeast Res, 2003 Jun, 3(4), 405 - 15 Molecular typing of clinical and environmental Cryptococcus neoformans isolates in the Brazilian state Rio Grande do Sul; Casali AK et al.; In Brazil, 4.5% of the AIDS-related opportunistic infections are caused by Cryptococcus neoformans . This pathogen is a ubiquitous environmental basidiomycetous encapsulated yeast, commonly found in soil and avian excreta . The present study investigates further the population structure of clinical and environmental C . neoformans isolates from south Brazil . One hundred five clinical and 19 environmental (pigeon excreta and Eucalyptus spp.) isolates from the Brazilian state Rio Grande do Sul were characterized based on morphological, biochemical, molecular and serological data . The majority of the clinical and environmental isolates analyzed belonged to C . neoformans var . grubii serotype A (89.5 and 52.6%, respectively), were mating type alpha (98.1 and 94.7%, respectively) and were phospholipase-positive (94.3 and 73.7%, respectively) . PCR-fingerprinting with the microsatellite-specific primer M13 and the minisatellite-specific primer (GACA)(4) grouped the majority of the isolates into the molecular type VNI (89.5 of the clinical and 52.6% of the environmental isolates) . Our results add considerable new information to the few available data on ecology, molecular biology and epidemiology of C . neoformans in the southern region of Brazil. FEMS Yeast Res, 2003 Jun, 3(4), 401 - 4 ATP leakage from yeast cells treated by extracellular glycolipids of Pseudozyma fusiformata; Kulakovskaya TV et al.; The ustilaginaceous yeast Pseudozyma fusiformata secreted glycolipids which were lethal to many yeasts and fungi more active at pH of about 4.0, and in the temperature range of 20-30 degrees C . Purified glycolipids enhanced non-specific permeability of the cytoplasmic membrane in sensitive cells, which resulted in ATP leakage and susceptibility of the cells to staining with bromocresol purple . Cells of Saccharomyces cerevisiae lost the ability to acidify the medium . Basidiomycetous yeasts were more sensitive to the glycolipids than ascomycetous ones . The minimal effective glycolipid concentration was 0.13 and 0.26 mg ml(-1) for Cryptococcus terreus and Filobasidiella neoformans, while for Candida albicans and Saccharomyces cerevisiae it was 1.0 and 1.6 mg ml(-1). Clin Infect Dis, 2003 May 15, 36(10), 1329 - 31 Epub 2003 May 02. Discontinuation of secondary prophylaxis for cryptococcal meningitis in human immunodeficiency virus-infected patients treated with highly active antiretroviral therapy: a prospective, multicenter, randomized study; Vibhagool A et al.; A prospective, multicenter, randomized study was conducted with human immunodeficiency virus (HIV)-infected patients who were successfully treated for acute cryptococcal meningitis, were receiving secondary prophylaxis, and were naive for antiretroviral therapy . Patients were randomized to continue or discontinue secondary prophylaxis when the CD4 cell count had increased to >100 cells/microL and an undetectable HIV RNA level had been sustained for 3 months . At a median of 48 weeks after randomization, there were no episodes of cryptococcal meningitis in either group. Expert Opin Pharmacother, 2003 May, 4(5), 807 - 23 Caspofungin: first approved agent in a new class of antifungals; Johnson MD et al.; Caspofungin (Cancidas, Merck & Co . Inc.) is the first echinocandin antifungal agent to gain FDA-approval for use in the US . It has excellent clinical activity against Candida spp . and Aspergillus spp . but lacks significant activity against Cryptococcus neoformans . Caspofungin may have some activity against dimorphic fungi such as Histoplasma capsulatum and Coccidioides immitis, but no clinical data is available for treatment of these infections . Caspofungin has demonstrated poor activity against most filamentous fungi in vitro . Several clinical trials have demonstrated its efficacy in the treatment of oropharyngeal, oesophageal and invasive candidiasis, as well as invasive aspergillosis . As a result of caspofungin's unique mechanism of action, and the high morbidity and mortality of invasive fungal infections, there is considerable interest in using this new antifungal agent as part of a combination antifungal therapy . In vitro studies and small case series indicate that caspofungin does not appear to be antagonistic when combined with other antifungals, such as itraconazole, voriconazole or amphotericin B against Aspergillus spp . Caspofungin exerts concentration-dependent killing effects in many different in vitro and animal models of disseminated fungal infection . The usual daily dose is 50 mg/day i.v . following a 70 mg i.v . loading dose . However, higher caspofungin doses have been safely administered and up to 70 mg/day can be administered for patients who fail to respond to lower doses . Caspofungin has an excellent safety profile with reduced toxicities, compared to other licensed antifungal agents . Fever, thrombophlebitis, headache and liver enzyme elevations were the most common drug-related side effects reported in clinical trials so far . Additional data are needed to document its safety in long-term use, and with higher doses in patients with invasive fungal infections . Caspofungin is a promising agent as first-line therapy for invasive candidiasis, and as salvage therapy for invasive aspergillosis . However, more clinical data are needed to define its role as primary therapy for invasive aspergillosis, and its role in combination antifungal therapy. Kansenshogaku Zasshi, 2003 Apr, 77(4), 227 - 34 {Comparison of four diagnostic methods using clinical blood by measuring (1-->3)-beta-D-glucan}; Moro H et al.; Using Amebocyte lysate of horseshoe crab to measure (1-->3)-beta-D-glucan specifically, a component of the cell wall, several kinds of diagnostic methods for deep mycosis are in practical use in Japan . However, the most important problem is that the judgment of positive or negative is method dependent . To elucidate the cause of the problem, each measurement value of the identical sample by four methods, beta-Glucan Test Maruha (MARUHA), beta-Glucan Test Wako (WAKO) . FUNGITEC G Test (FUNGITEC-G) and FUNGITEC G Test MK (FUNGITEC-MK) was compared with the clinical data using 119 cases and 289 tests . Each case was divided into three groups; proven fungal infection, probable fungal infection and non-fungal infection . The negative cases for all the methods tested in the groups of proven fungal infection and probable fungal infection were allergic bronchopulmonary aspergillosis and cryptococcosis, and that for all the methods tested except FUNGITEC-MK method in the group was pulmonary aspergilloma . It seems that these cases cannot be detected correctly by only measuring (1-->3)-beta-D-glucan . On the other hand, the ratio of false positive, positive for non-fungal infection group was high in the case of FUNGITEC-MK . About 23% against the total case was positive for FUNGITEC-MK method, but negative for MARUHA, WAKO, and FUNGITEC-G methods . Although the difference of the sensitivity among four methods was not observed, the specificity, the diagnostic efficiency, and the positive predictive value of FUNGITEC-MK method were remarkably lower than those of the other methods due to false positive measurement . In conclusion, MARUHA, WAKO and FUNGITEC-G except FUNGITEC-MK is not method dependent . It is suggested that FUNGITEC-MK detects non-specific reaction resulted in false positive. Curr Opin Infect Dis, 2003 Apr, 16(2), 91 - 6 Melanin and fungi; Gomez BL et al.; PURPOSE OF REVIEW: Production of melanin has been associated with virulence in diverse microorganisms . Melanization of fungi has been noted for many years in predominantly subcutaneous infections such as chromoblastomycosis and more recently most extensively studied in a yeast causing systemic infection, Cryptococcus neoformans . Pigmented fungi are increasingly important human pathogens and currently available antifungals are often sub-optimal for serious infections . This review focuses on recent publications on melanin in fungi with particular reference to the role of melanin in virulence, protection against antifungal drugs, and promoting survival in the environment . RECENT FINDINGS: Inhibition of melanin production by C . neoformans can prolong survival of lethally infected mice . In contrast, melanin in C . neoformans and Histoplasma capsulatum yeast cells can bind amphotericin B and caspofungin, thereby reducing the fungicidal affects of these drugs . H . capsulatum and Paracoccidioides brasiliensis have only recently been shown to produce melanin in vitro and during infection . Additionally, melanin derived from melanized C . neoformans yeast and Aspergillus niger conidia can activate complement, which may modify immune responses to infection . Studies on C . neoformans laccase have revealed that the enzyme is located on the cell wall, which may allow for interactions with the host . Melanization reduces the susceptibility of C . neoformans to enzymatic degradation and toxicity from a heavy metal, which may afford protection to the fungus against similar insults in the environment . SUMMARY: Melanin has been referred to as 'fungal armor' due to the ability of the polymer to protect microorganisms against a broad range of toxic insults . Recent publications continue to reveal important contributions of melanin to survival of fungi in the environment and during infection. J Trop Pediatr, 2003 Apr, 49(2), 124 - 5 Cryptococcal meningitis in pediatric AIDS; Kaur R et al.; Cryptococcus neoformans is an infrequent cause of meningitis in children . We report two cases of HIV positive children of HIV non-reactive parents who were diagnosed as suffering from cryptococcal meningitis . Treatment with amphotericin B and flucytosine was instituted in both the children who recovered and are doing well. J Chin Med Assoc, 2003 Jan, 66(1), 19 - 26 Radiological manifestations of cryptococcal infection in central nervous system; Cheng YC et al.; BACKGROUND: Cryptococcus neoformans is one of the most common infectious agents causing central nervous system (CNS) infections in immunocompromised patients . Patients with CNS cryptococcosis commonly present with non-specific manifestations such as headache, fever, seizure and conscious disturbance . This article presents the computerized tomography scans (CT) and magnetic resonance images (MRI) findings of cryptococcal infection in CNS and displays the image difference between immunocompromised and non-immunocompromised patients . METHODS: CNS cryptococcal infection was found in 26 cases over the past 14 years in Taipei Veterans General Hospital, including 18 males and 8 females . Ten patients were immunocompromised . In all cases, the CNS cryptococcosis was diagnosed by positive cryptococcal antigen titer and India ink stain from cerebral spinal fluid (CSF) . Their clinical data and findings from CT and MRI were retrospectively studied . RESULTS: The most common CT manifestation is ventricular dilatation; other common findings include leptomeningeal enhancement, infarction (half of them revealed dilatation of Virchow-Robin spaces in subsequent MRI study) and normal appearance . Less common findings are ventriculitis and abscess formation . The most common MRI pattern is Virchow-Robin space dilatation and leptomeningeal enhancement . Other common appearance includes ventricular dilatation . Less common manifestations include cryptococcoma, hemorrhagic infarction and pontine abscess . There is no significant difference in image patterns between immunocompromised and non-immunocompromised patients (p > 0.05) . CONCLUSIONS: The most common pattern ofCNS cryptococcosis is ventricular dilatation in CT and Virchow-Robin space dilatation in MRI . MRI is more sensitive in detecting CNS cryptococcal infection like Virchow-Robin space dilatation and leptomeningeal enhancement . There is no significant pattern difference between immunocompromised and non-immunocompromised patients with CNS cryptococcosis. Infez Med, 2003 Mar, 11(1), 42 - 5 {Cutaneous cryptococcosis in an immunocompetent host}; Coppola C et al.; An immunocompetent 62-year-old woman presented with cutaneous lesions over her right arm and the hypogastrium . The diagnosis of cutaneous cryptococcosis was made on the basis of the demonstration and isolation of Cryptococcus neoformans from the aspirated pus . The patient responded to fluconazole treatment. Clin Infect Dis, 2003 May 1, 36(9), 1122 - 31 Epub 2003 Apr 22. Voriconazole treatment for less-common, emerging, or refractory fungal infections; Perfect JR et al.; Treatments for invasive fungal infections remain unsatisfactory . We evaluated the efficacy, tolerability, and safety of voriconazole as salvage treatment for 273 patients with refractory and intolerant-to-treatment fungal infections and as primary treatment for 28 patients with infections for which there is no approved therapy . Voriconazole was associated with satisfactory global responses in 50% of the overall cohort; specifically, successful outcomes were observed in 47% of patients whose infections failed to respond to previous antifungal therapy and in 68% of patients whose infections have no approved antifungal therapy . In this population at high risk for treatment failure, the efficacy rates for voriconazole were 43.7% for aspergillosis, 57.5% for candidiasis, 38.9% for cryptococcosis, 45.5% for fusariosis, and 30% for scedosporiosis . Voriconazole was well tolerated, and treatment-related discontinuations of therapy or dose reductions occurred for <10% of patients . Voriconazole is an effective and well-tolerated treatment for refractory or less-common invasive fungal infections. J Biol Chem, 2003 Jun 27, 278(26), 23639 - 47 Epub 2003 Apr 24. GWT1 gene is required for inositol acylation of glycosylphosphatidylinositol anchors in yeast; Umemura M et al.; Glycosylphosphatidylinositol (GPI) is a conserved post-translational modification to anchor cell surface proteins to plasma membrane in all eukaryotes . In yeast, GPI mediates cross-linking of cell wall mannoproteins to beta1,6-glucan . We reported previously that the GWT1 gene product is a target of the novel anti-fungal compound, 1-{4-butylbenzyl}isoquinoline, that inhibits cell wall localization of GPI-anchored mannoproteins in Saccharomyces cerevisiae (Tsukahara, K., Hata, K., Sagane, K., Watanabe, N., Kuromitsu, J., Kai, J., Tsuchiya, M., Ohba, F., Jigami, Y., Yoshimatsu, K., and Nagasu, T . (2003) Mol . Microbiol . 48, 1029-1042) . In the present study, to analyze the function of the Gwt1 protein, we isolated temperature-sensitive gwt1 mutants . The gwt1 cells were normal in transport of invertase and carboxypeptidase Y but were delayed in transport of GPI-anchored protein, Gas1p, and were defective in its maturation from the endoplasmic reticulum to the Golgi . The incorporation of inositol into GPI-anchored proteins was reduced in gwt1 mutant, indicating involvement of GWT1 in GPI biosynthesis . We analyzed the early steps of GPI biosynthesis in vitro by using membranes prepared from gwt1 and Deltagwt1 cells . The synthetic activity of GlcN-(acyl)PI from GlcN-PI was defective in these cells, whereas Deltagwt1 cells harboring GWT1 gene restored the activity, indicating that GWT1 is required for acylation of inositol during the GPI synthetic pathway . We further cloned GWT1 homologues in other yeasts, Cryptococcus neoformans and Schizosaccharomyces pombe, and confirmed that the specificity of acyl-CoA in inositol acylation, as reported in studies of endogenous membranes (Franzot, S . P., and Doering, T . L . (1999) Biochem . J . 340, 25-32), is due to the properties of Gwt1p itself and not to other membrane components. Infez Med, 2000, 8(3), 167 - 172 {Current epidemiological and clinical features meningitis in a northern Italian area}; De Carlo A et al.; Objectives: To study etiological, epidemiological and clinical features of 97 cases of acute meningitis . Methods: Ninety-seven cases of acute meningitis were examined in adult HIV-negative patients admitted to the Infectious Diseases Unit of the Azienda Ospedale-Universita S . Anna in Ferrara . Demographic, etiological, epidemiological and clinical data were analyzed . Results: All cases were divided into two groups according to the macroscopic aspect of cerebrospinal fluid (CSF): purulent CSF (50 cases) or non-purulent CSF (47 cases) . Purulent CSF meningitis more frequently affected male patients (64% vs 47%) and older patients (average 52 vs 44 years) . The main epidemiological features in both groups were underlying bacterial diseases (i.e . otomastoiditis and/or sinusitis in 50% of pneumococcal meningitis) and iatrogenic immunodeficiency . From a clinical point of view the following alterations in the state of consciousness (stupor, confusion and coma) were most frequently found in purulent meningitis . The following non purulent forms of meningitis were diagnosed: 5 tubercular, 3 viral infections, 2 by Listeria monocytogenes, 1 by Entoameba histolytica, 1 by Cryptococcus neoformans and 35 (74,4%) unknown causes . Purulent meningitis were: 20 (40%) Streptococcus pneumoniae, 10 Neisseria meningitidis, 3 Staphylococcus aureus, 2 Escherichia coli, 1 Haemophilus influenzae and 1 Pseudomonas aeruginosa; 13 cases were unidentified . From 1989 to 1993 and from 1994-98 both groups of meningitis increased; respectively from 17 to 30 cases for non-purulent meningitis and from 18 to 32 cases for purulent meningitis . Meningitis due to Streptococcus pneumoniae increased from 27.7% to 46.8% during the period 1994-98 . Conclusions: The study shows the high incidence of pneumococcal meningitis, during 1994-98, because a large number of patients with sinusitis and otomastoiditis were observed . The incidence of meningococcal meningitis appears stable . These data confirm the importance of timely diagnosis and correct therapy for such infections with reserved prognosis. Infez Med, 2000, 8(3), 134 - 138 {Effects of the introduction of highly active antiretroviral therapy (HAART) on the temporal trend of diagnosis of AIDS and AIDS-defining disorders}; Manfredi R; Objectives: to assess the features of AIDS and AIDS-defining events during the highly active antiretroviral therapy (HAART) era, compared with those observed before the introduction of HAART . Methods: the epidemiological, clinical and laboratory characteristics of 72 consecutive patients with AIDS diagnosed from 1997 to 1999, were compared to those of 144 subjects randomly selected from 436 AIDS patients diagnosed from 1985 and 1995, in a case control study . Results: a sharp drop in AIDS notifications was observed after the introduction of HAART, in association with an increase in mean patient age, and heterosexual transmission of HIV infection . A tendency to reduced frequency of Cytomegalovirus disease, cryptococcosis, mycobacteriosis, cryptosporidiosis and HIV encephalopathy was also noticed, with a slight proportion increase in pneumocystosis, esophageal candidiasis, wasting syndrome, tuberculosis and lymphoma . Non-infectious complications showed a slighly increased frequency during the HAART era, while a mild parallel decrease in overall opportunistic infections was observed . A considerable trend to an increased mean CD4+ lymphocyte count at diagnosis was found during the HAART era for most AIDS-related illnesses (significant for Candida esophagitis, wasting syndrome and tuberculosis) (p<.04) . Only seven of the 72 patients with AIDS recognized since 1997 received HAART prior to diagnosis, while the remaining 65 cases HIV infection was detected concurrently with an AIDS-defining event (40 cases), or HAART was refused or carried out with poor adherence (25 patients) Conclusions: although remarkable changes of AIDS features of AIDS occurred after the widespread use of HAART, early diagnosis and treatment of HIV infection, and careful monitoring of the spectrum of AIDS-related diseases are strongly warranted in order to update and plan future management strategies. Kansenshogaku Zasshi, 2003 Mar, 77(3), 150 - 7 {A clinical study of cryptococcal meningitis--sequential changes of cryptococcal antigen titers}; Kishi K et al.; We reviewed the clinical manifestations, sequential changes in cryptococcal antigen titers in serum and cerebrospinal fluid (CSF), and the antifungal drug susceptibility of Cryptococcus neoformans in three patients with cryptococcal meningitis between 1996 and 2000 . Cryptococcal antigen titers were measured using the latex agglutination method with Pastrex Cryptococcus (Fuji Mebio, Tokyo) and Serodirect Cryptococcus (Eiken Chemical, Tokyo) . The underlying systemic diseases in the three patients were liver cirrhosis, non-Hodgkin's lymphoma associated with miliary tuberculosis, and malignant thymoma associated with systemic lupus erythymatosus . The CSF samples showed positive indian ink staining in two of the three patients and C . neoformans was cultured from all three . The cryptococcal antigen titers in serum were higher than those in the CSF . The serum and CSF cryptococcal antigen titers measured by Serodirect Cryptococcus were higher than those measured by Pastrex Cryptococcus . The maximum titers of antigen in serum and CSF measured by Serodirect Cryptococcus were greater than 1,024 in all three patients . The treatment regimens used for the three patients were amphotericin-B (AMPH-B) and flucytosine (5-FC), fluconazole (FLCZ) and intrathecal AMPH-B, FLCZ and 5-FC, and intrathecal AMPH-B, respectively . The antigen titers in serum and CSF decreased after treatment in all three patients . The antigen titers decreased slowly over 7.3 months in the most seriously ill patient who had non-Hodgkin's lymphoma associated with miliary tuberculosis . The time between the beginning of treatment and CSF cryptococal antigen titers falling to less than 8 was 1.7 to 7.3 months in the three patients, but the serum titers did not decrease to less than 8 during this period . The minimum inhibitory concentration was 0.06-0.25 microgram/ml for AMPH-B, 4-8 micrograms/ml for 5-FC, 2-8 micrograms/ml for FLCZ, 0.125-0.5 microgram/ml for miconazole and 0.03-0.125 microgram/ml for itraconazole . The measurement of sequential changes in cryptococcal antigen titers in serum and CSF was useful for evaluating the response to treatment. Microbes Infect, 2003 Apr, 5(4), 285 - 90 Identification of metabolites of importance in the pathogenesis of pulmonary cryptococcoma using nuclear magnetic resonance spectroscopy; Himmelreich U et al.; Primary lung infection with Cryptococcus neoformans is characterised by circumscribed lesions (cryptococcomas) . To identify cryptococcal and/or host products of importance in pathogenesis, we applied proton nuclear magnetic resonance (NMR) spectroscopy, which identifies mobile compounds present in complex mixtures, to experimental pulmonary cryptococcomas from rats . Magnetic resonance experiments were performed on cryptococcomas (n = 10) and healthy lungs (n = 8) . Signal assignment to key metabolites was confirmed by homo-nuclear and hetero-nuclear NMR correlation spectroscopy . Cryptococcal metabolites, dominating spectra from cryptococcomas included the stress protectants, trehalose and mannitol, acetate, and in some animals, ethanol . Glycerophosphorylcholine was also abundant in cryptococcomas, consistent with hydrolysis of phospholipids in vivo by the cryptococcal enzyme, phospholipase B (PLB) . PLB has been identified by molecular studies as a cryptococcal virulence determinant . We propose that PLB secreted by cryptococci promotes tissue invasion by hydrolysing host phospholipids, such as dipalmitoyl phosphatidyl choline, which is abundant in pulmonary surfactant, and lung cell membrane phospholipids . Our results confirm the utility of NMR spectroscopy in studies of microbial pathogenesis. Infect Immun, 2003 May, 71(5), 2868 - 75 Antigenic and biological characteristics of mutant strains of Cryptococcus neoformans lacking capsular O acetylation or xylosyl side chains; Kozel TR et al.; Cryptococcus neoformans is surrounded by an antiphagocytic polysaccharide capsule whose primary constituent is glucuronoxylomannan (GXM) . Three prominent structural features of GXM are single xylosyl and glucuronosyl side chains and O acetylation of the mannose backbone . Isogenic pairs of O-acetyl-positive and O-acetyl-negative strains (cas1 Delta) as well as xylose-positive and xylose-negative strains (uxs1 Delta) of serotype D have been reported . The cas1 Delta strains were hypervirulent, and the uxs1 Delta strains were avirulent . The goal of this study was to examine the effects of the cas1 Delta and uxs1 Delta mutations on the following: (i) binding of anti-GXM monoclonal antibodies (MAbs) in capsular quellung reactions, (ii) activation of the complement system and binding of C3, (iii) phagocytosis by neutrophils, and (iv) clearance of GXM in vivo . The results showed that loss of O acetylation produced dramatic changes in the reactivities of five of seven anti-GXM MAbs . In contrast, loss of xylosylation produced a substantive alteration in the binding behavior of only one MAb . O-acetyl-negative strains showed no alteration in activation and binding of C3 from normal serum . Xylose-negative strains exhibited accelerated kinetics for C3 deposition . Loss of O acetylation or xylosylation had no effect on phagocytosis of serum-opsonized yeast cells by human neutrophils . Finally, loss of O acetylation or xylosylation altered the kinetics for clearance of GXM from serum and accumulation of GXM in the liver and spleen . These results show that O acetylation and/or xylosylation are important for binding of anti-GXM MAbs, for complement activation, and for tissue accumulation of GXM but do not impact phagocytosis by neutrophils. Genetics, 2003 Apr, 163(4), 1315 - 25 Mitochondria are inherited from the MATa parent in crosses of the basidiomycete fungus Cryptococcus neoformans; Yan Z et al.; Previous studies demonstrated that mitochondrial DNA (mtDNA) was uniparentally transmitted in laboratory crosses of the pathogenic yeast Cryptococcus neoformans . To begin understanding the mechanisms, this study examined the potential role of the mating-type locus on mtDNA inheritance in C . neoformans . Using existing isogenic strains (JEC20 and JEC21) that differed only at the mating-type locus and a clinical strain (CDC46) that possessed a mitochondrial genotype different from JEC20 and JEC21, we constructed strains that differed only in mating type and mitochondrial genotype . These strains were then crossed to produce hyphae and sexual spores . Among the 206 single spores analyzed from six crosses, all but one inherited mtDNA from the MATa parents . Analyses of mating-type alleles and mtDNA genotypes of natural hybrids from clinical and natural samples were consistent with the hypothesis that mtDNA is inherited from the MATa parent in C . neoformans . To distinguish two potential mechanisms, we obtained a pair of isogenic strains with different mating-type alleles, mtDNA types, and auxotrophic markers . Diploid cells from mating between these two strains were selected and 29 independent colonies were genotyped . These cells did not go through the hyphal stage or the meiotic process . All 29 colonies contained mtDNA from the MATa parent . Because no filamentation, meiosis, or spore formation was involved in generating these diploid cells, our results suggest a selective elimination of mtDNA from the MATalpha parent soon after mating . To our knowledge, this is the first demonstration that mating type controls mtDNA inheritance in fungi. FEMS Yeast Res, 2001 Jul, 1(2), 103 - 9 Description of Bullera kunmingensis sp . nov., and clarification of the taxonomic status of Bullera sinensis and its synonyms based on molecular phylogenetic analysis; Bai FY et al.; A ballistoconidium-forming yeast strain, CH 2.506, isolated from a semi-dried leaf of Parthenocissus sp . collected near Kunming City in Yunnan, China, was shown to be closely related to the non-ballistoconidium-forming species Cryptococcus luteolus (Saito) C.E . Skinner and the ballistoconidium-forming species Bullera sinensis Li by molecular phylogenetic analysis based on 18S rDNA sequencing . This strain was demonstrated to represent a distinct undescribed yeast species by internal transcribed spacer (ITS) region sequence and G+C content comparison and DNA-DNA relatedness, for which the name Bullera kunmingensis sp . nov . is proposed . Meanwhile, the taxonomic relationships among Bullera sinensis and its synonyms B . derxii Nakase & Suzuki and B . alba (Hanna) Derx var . lactis Li, were clarified on the basis of molecular phylogenetic analysis and DNA-DNA reassociation . B . derxii was confirmed to be conspecific with B . sinensis, while B . alba var . lactis was shown to represent a variety of B . sinensis . A new combination, Bullera sinensis Li var . lactis (Li) Bai, Takashima et Nakase, is therefore proposed . Comparative analysis of different types of molecular criteria employed in the present study suggested that when inferring phylogenetic relationships among sibling taxa, sequence data from ITS regions should be interpreted with caution. FEMS Yeast Res, 2001 Dec, 1(3), 221 - 4 Rapid methods to extract DNA and RNA from Cryptococcus neoformans; Bolano A et al.; Extraction of nucleic acids from the pathogenic yeast Cryptococcus neoformans is normally hampered by a thick and resistant capsule, accounting for at least 70% of the whole cellular volume . This paper presents procedures based on mechanical cell breakage to extract DNA and RNA from C . neoformans and other capsulated species . The proposed system for DNA extraction involves capsule relaxation by means of a short urea treatment and bead beating . These two steps allow a consistent extraction even from strains resistant to other procedures . Yield and quality of DNA obtained with the proposed method were higher than those obtained with two earlier described methods . This protocol can be extended to every yeast species and particularly to those difficult to handle for the presence of a capsule . RNA purification is accomplished using an original lysing matrix and the FastPrep System (Bio101) after a preliminary bead beating treatment . Yields range around 1 mg RNA from 15 ml overnight culture (10(9) cells), RNA appears undegraded, making it suitable for molecular manipulations. FEMS Yeast Res, 2001 Dec, 1(3), 213 - 20 Genetic diversity in the red yeast Cryptococcus hungaricus and its phylogenetic relationship to some related basidiomycetous yeasts; Gacser A et al.; Cryptococcus hungaricus is a basidiomycetous yeast with the abilities to synthesize carotenoid pigments and to grow under psychrophile conditions . Six C . hungaricus strains have been isolated so far from different habitats . In this study we wished to clarify the relationships amongst them . Morphological and physiological characters, mitochondrial DNA restriction profiles, and the presence of mycoviruses were examined . Internal transcribed spacers together with the 5.8S rDNA, the D1/D2 region of 26S rDNA, and partial sequences of the 18S rRNA gene were also analysed . On the basis of the phylogenetic analyses the type strain CBS 4214(T) together with four other C . hungaricus isolates were closely related to Bullera armeniaca and Bullera crocea, while strain CBS 6569 was much more similar to Cystofilobasidium than to the other C . hungaricus isolates. FEMS Yeast Res, 2002 Dec, 2(4), 551 - 61 Isolation and characterisation of the phospholipase B gene of Cryptococcus neoformans var . gattii; Latouche GN et al.; Cryptococcus neoformans var . gattii (serotypes B and C) is a human pathogen, ecologically, biochemically, clinically and genetically different from C . neoformans var . grubii (serotype A) and C . neoformans var . neoformans (serotype D) . The phospholipase B (PLB1) gene from serotypes B and C was isolated and characterised . It resembled the serotype A and D genes, with an overall sequence homology of more than 85% . The respective open reading frames were 2236 bp (serotype B) and 2239 bp (serotype C) in length . Each contained six introns and encoded a 68-kDa protein destined for secretion . PLB1 was located on the second smallest chromosome in both serotypes . Gene expression, measured as mRNA, was not regulated by temperature, pH or exogenous nutrients. FEMS Yeast Res, 2003 Mar, 3(1), 63 - 8 Double-stranded RNA viruses in a mycocinogenic strain of Cystofilobasidium infirmominiatum; Golubev WI et al.; The viral particles (about 30 nm in diameter) that contain dsRNAs (2.0 and 6.3 kbp) encapsidated by a coat of protein were detected in a mycocin-secreting strain of Cystofilobasidium infirmominiatum isolated from plants in an oak forest (Moscow region) . The mycocin with a molecular mass above 15 kDa is fungicidal (maximum activity at pH 4.5) and active mainly against some species of the Cystofilobasidiales and Filobasidiales ('Cryptococcus aerius' clade) . Curing by incubation at elevated temperature resulted in the concomitant loss of dsRNAs and mycocinogenic activity, and cured derivatives became sensitive to the mycocin produced by the parent strain. J Gen Appl Microbiol, 2002 Dec, 48(6), 345 - 55 Bullera taiwanensis sp nov and Bullera formosensis sp nov, two new ballistoconidium-forming yeast species isolated from plant leaves in Taiwan; Nakase T et al.; Two strains of ballistoconidiogenous yeasts that contain xylose and form Q-10 ubiquinone were isolated from plant leaves collected in Taiwan and were found to represent two new species . The taxonomic properties of both coincide with the genus Bullera so they are described as Bullera taiwanensis sp . nov . and Bullera formosensis sp . nov . In a phylogenetic tree based on the nucleotide sequences of 18S ribosomal DNAs, these two species are distant from the clusters where the remaining members of the genus Bullera are located, i.e., Bullera taiwanensis is located in the Filobasidium lineage (Filobasidiales clade) and Bullera formosensis is located in the Cryptococcus humicola-Trichosporon lineage (Trichosporonales clade). J Clin Microbiol, 2003 Apr, 41(4), 1736 - 7 Detection of single cells of Cryptococcus neoformans in clinical samples by solid-phase cytometry; Bauters TG et al.; A method based on solid-phase cytometry for the detection and enumeration of single cells of Cryptococcus neoformans in serum and cerebrospinal fluid is described . Both viable and nonviable cells are detected by using fluorescence viability labeling and immunofluorescence . This 30-min procedure has a detection limit of 3 to 6 cells per ml. Lancet, 2003 Apr 12, 361(9365), 1267 - 8 S-adenosylmethionine concentrations in diagnosis of Pneumocystis carinii pneumonia; Skelly M et al.; Pneumocystis carinii is unable to synthesise S-adenosylmethionine and thus scavenges this intermediate . We aimed to test whether measurement of concentrations of this metabolic intermediate in plasma could provide a new method for rapid diagnosis of Pneumocystis carinii pneumonia (PCP) . We measured S-adenosylmethionine plasma concentrations in 12 healthy controls, 16 patients with confirmed or suspected PCP, and 36 patients with other infections . Median concentration in healthy controls was 106 nmol/L (range 86-128), but the protein was undetectable in eight patients with histologically proven and seven with suspected PCP, and was 8 nmol/L in another confirmed case (p<0.0001) . In 36 patients with other infections, S-adenosylmethionine concentrations were much the same as in controls: 18 had bacterial pneumonia, two tuberculosis, five cryptococcal meningitis, three had other infections, and eight had asymptomatic HIV-1 infection . After treatment for PCP, S-adenosylmethionine concentrations rose rapidly in all but one patient who died of the disease . Measurement of plasma S-adenosylmethionine concentrations could prove useful for diagnosis of PCP and assessment of patients' response to treatment. Nihon Kokyuki Gakkai Zasshi, 2003 Jan, 41(1), 30 - 4 {Pulmonary cryptococcosis combined with pulmonary tuberculosis}; Kishi K et al.; We report a rare combination of pulmonary cryptococcosis and pulmonary tuberculosis in a diabetic patient . A 63-year-old man was admitted to our hospital in January 2002 for evaluation of an abnormal chest radiograph . In 1999, the abnormality had first been detected by mass screening radiography . In 2000, an abnormality in a chest radiograph was again detected in a mass screening, and he visited another hospital . A chest CT scan revealed a cavitating lesion and several nodules in the left lower lobe . Fiberoptic bronchoscopy was performed, but was not diagnostic . The patient was referred to our hospital . He had a history of diabetes mellitus starting in 1984 . The chest CT scan revealed solid nodules in the left lower lobe and several micronodules in both upper lobes . Video-assisted thoracoscopic surgery was performed and specimens were obtained from the left S8 and left S1 + 2 . Histologically, cryptococci were detected in the resected left S8 . In addition, mycobacterium tuberculosis was cultured from the resected left S1 + 2 . A diagnosis of combined pulmonary cryptococcal and tuberculous infections was made and treatment with itraconazole, isoniazid sodium methansulfonate, rifampicin, and ethambutol hydrochloride was given. J Korean Med Sci, 2003 Apr, 18(2), 149 - 57 Neurologic complications of human immunodeficiency virus-type 1 infection; Kim HJ et al.; A wide variety of neurologic complications associated with human immunodeficiency virus-type 1 (HIV-1) infection result from HIV-1 itself or secondarily related to immunosuppression . In Korea, the number of HIV-1 seropositive populations is increasing, but little has been known about the neurologic complications of HIV-1 infection . To investigate the neurologic complications in HIV-1 infected Korean patients, we performed a cross-sectional study in consecutive admissions to the Seoul National University Hospital between March 1998 and June 1999 . Thirty-four HIV-1 seropositive patients were included . As a result, a total of 26 HIV-1 related neurologic complications were identified from 17 patients . Among them, 10 patients showed cognitive/motor abnormalities: 3 HIV-1-associated dementia and 7 possible HIV-1-associated minor cognitive/motor disorder . Neuromuscular complications were found in 10 patients: 9 distal symmetric polyneuropathy, and 1 possible chronic inflammatory demyelinating polyradiculoneuropathy . In 3 patients with focal brain lesions, 2 were presumptively diagnosed as having primary CNS lymphoma, and 1 as having progressive multifocal leukoencephalopathy in the posterior fossa, based on history, clinical findings, serology, radiological appearances, and response to empirical therapy . Other complications included cryptococcal meningitis and only soft neurologic signs without any neurologic disease . Most of these complications (88%) occurred in the advanced stage of infection. Mycoses, 2002, 45 Suppl 3, 61 - 4 {IWhich are the conditions for Cryptococcus neoformans var . neoformans-strains from avian excrements as a cause for human infections?}; Kielstein P et al.; Detection of antigen factors of Cryptococcus with factor sera in slide agglutination confirms diagnosis of species and varieties of Cryptococcus neoformans (Cr . n) . This method is important in investigations of sources of infections . Serotype D strains of Cr . neoformans were detected in pigeon breedings from Thuringia exclusively . Because of that an essential difference exists in comparison to human isolates in Germany and strains from breeding stocks of companion birds in Thuringia where serotype A strains are predominant in pet birds and in human infections . Using different primers in PCR fingerprinting Cr . neoformans isolates can be assigned to serotypes A, B, C and D and to varieties Cr . neoformans neoformans and Cr . neoformans gattii (primer FM 1) . On the other hand, genetic heterogeneity of Cr . neoformans strains is detectable within the serotypes A and D (primer 60-26) . This genetic heterogeneity can be demonstrated in investigations by Fourier Transform Infrared (FTIR) spectroscopy, too . Isolated Cr . neoformans strains from pigeons (serotype D) could be divided into 3 and from pet birds (serotype A) into 2 different clusters by FTIR spectroscopy . It is important to take into account heterogeneity of strains within serotypes for determination of infection chains of human disease. Mycoses, 2002, 45 Suppl 3, 56 - 60 The first echinocandin: caspofungin; Cornely OA et al.; The antifungal agent caspofungin is the first echinocandin that has been approved in the US and in Europe for treatment of invasive aspergillosis in adult patients who are refractory to or intolerant of conventional amphotericin B, its lipid-based formulations, and/or itraconazole . It is given as a 70 mg loading dose and a 50 mg daily maintainance dose as a one hour infusion . Due to low intestinal absorption an oral formulation has not been developed . Caspofungin is active against Candida spp . and Aspergillus spp . by inhibition the synthesis of beta-(1,3)-D-glucan, a cell wall component . The drug is inactive against Cryptococcus spp., Fusarium spp., Trichosporon spp., Rhizopus spp., and Pseudoallescheria spp . In invasive aspergillosis caspofungin resulted in higher response rates compared to a historic control under standard therapy . Efficacy data on persistently febrile neutropenic patients are pending . In several multicenter randomised double blind trials on candida infections caspofungin proved to be at least non-inferior to standard therapies . Reports of combination therapy or highly effective antifungal treatment (HEAT) in limited patient numbers are promising . New trials of combination therapy are warranted. Mycoses, 2002, 45 Suppl 3, 42 - 7 {Voriconazole - applications and perspectives}; Ruhnke M; Voriconazole (Vfend) is a new broadspectrum antifungal agent belonging to the group of triazole drugs . In vitro and in vivo efficacy was demonstrated against a large variety of yeasts with excellent activity against all Candida species but as well against Cryptococcus neoformans . Furthermore, voriconazole has shown excellent activity against many moulds in particular against Aspergillus species, but endemic fungi such as Histoplasma capsulatum are in the spectrum as well . Clinical efficacy was demonstrated in several large phase II/III studies in diseases such as oral and oesophageal candidosis, acute invasive aspergillosis or chronic invasive aspergillosis . New adverse events such as visual disturbancies has been described together with the use of voriconazole, but the majority of adverse events are similar to other triazole drugs and in particular not life-threatening . With the introduction of voriconazole a great progress in the therapy of invasive fungal infections was achieved . In the therapy of invasive aspergillosis, voriconazole is significantly more effective compared to amphotericin B desoxycholate. Neurol Clin, 2003 Feb, 21(1), 193 - 219 Nervous system infections in patients with cancer; Pruitt AA; The diagnostic approach to the patient with cancer with suspected CNS infection depends on an analysis of the patient's immune defect, the time course of development of manifestations of infection, and the type of clinical syndrome with supportive evidence for a specific diagnosis coming from laboratory and neuroradiographic data . Most patients with CNS infections can be grouped into those with signs of meningitis or meningoencephalitis and those with focal mass lesions . A smaller group presents with stroke-like onset . Except for the group with strokes, those with focal deficits usually present in a more indolent fashion, whereas those with meningitis and encephalitis present more acutely {63} . Patients with B-lymphocyte dysfunction are susceptible to encapsulated bacterial pathogens . Patients with T-lymphocyte impairment develop CNS infections that are caused by intracellular pathogens, particularly viruses (HSV, JC, CMV, HHV-6), Nocardia, Aspergillus, and Toxoplasma . Many noninfectious entities, such as drug treatment complications, radiation effects, recurrent tumor, and paraneoplastic syndromes, can mimic CNS infections . Although cryptococcosis, bacterial meningitis, and some viral infections are easily diagnosed from Gram's stain, culture, or PCR, patients with mass lesions may require tissue biopsy to confirm diagnosis . Patients with cancer differ from normal hosts in the distribution of pathogens, and there is a wider range of differential diagnostic issues, both infectious and noninfectious, for the relatively few clinical syndromes that present as potential CNS infections. J Heart Lung Transplant, 2003 Apr, 22(4), 478 - 83 Anasarca caused by Cryptococcus neoformans after heart transplantation; Molino D et al.; Cryptococcosis is an opportunistic infection caused by the yeast-like fungus Cryptococcus neoformans . The infection predominantly strikes patients with cell-mediated immunodeficiency, that is, patients with organ transplants, leukemia, lymphoma, AIDS, and those receiving steroids or immunosuppressants . We describe a patient with skin lesions and anasarca secondary to intestinal infection from Cryptococcus neoformans after heart transplantation . We based diagnosis on histologic examination of the cutaneous lesions and of the duodenal mucosa . This case demonstrates that in immunosuppressed patients with anasarca of unknown origin, a diagnosis of intestinal opportunistic infection also should be considered. Microbiology, 2003 Apr, 149(Pt 4), 1041 - 9 Regulation of cytochrome c oxidase subunit 1 (COX1) expression in Cryptococcus neoformans by temperature and host environment; Toffaletti DL et al.; In the study of differential gene expression of Cryptococcus neoformans, a transcript of COX1 (cytochrome oxidase c subunit 1) was identified in a serotype A strain . The transcript was upregulated at 37 degrees C compared to 30 degrees C and expressed by yeasts infecting the central nervous system . Northern analysis of COX1 from the serotype A strain revealed two polycistronic transcripts, a temperature-upregulated 2.3 kb transcript and a 1.9 kb transcript that was not affected by temperature . In contrast, COX1 in a serotype D strain showed only a 1.9 kb polycistronic transcript plus a 1.6 kb monocistronic message, and temperature had no effect on the transcripts . The sequence of COX1 revealed similar coding regions between the two strains, but the serotype D strain had five introns whereas no introns were found in the serotype A strain . The serotype D strain had reduced growth rates compared to the serotype A strain at 37 degrees C, but in an AD hybrid strain the serotype D COX1 gene could support efficient high temperature growth . These studies have revealed mitochondrial molecular differences between serotype A and D strains which show evolutionary divergence . It will be important to determine whether differences in mitochondrial structure and function can influence cryptococcosis. Scand J Infect Dis, 2003, 35(1), 68 - 70 Concomitant tuberculous and cryptococcal thyroid abscess in a human immunodeficiency virus-infected patient; Kiertiburanakul S et al.; Suppurative thyroiditis is a rare condition caused mostly by Staphylococcus aureus and streptococci . Both tuberculous and cryptococcal thyroid abscess are even rarer . The incidence of extrapulmonary forms of tuberculosis and cryptococcosis has increased in areas with a high prevalence of human immunodeficiency virus (HIV) infection . A case is reported of dual infection by M . tuberculosis and Cryptococcus neoformans presenting as a thyroid abscess in a 32-y-old woman with symptomatic HIV infection . Atypical presentations of both tuberculosis and cryptococcosis should be considered in areas with a high incidence of these diseases. Appl Environ Microbiol, 2003 Apr, 69(4), 2080 - 6 PCR-restriction fragment length polymorphism analysis of the phospholipase B (PLB1) gene for subtyping of Cryptococcus neoformans isolates; Latouche GN et al.; Cryptococcus neoformans is a pathogenic yeast that is currently divided into three varieties, five serotypes, and eight molecular types . The following report describes the use of PCR-restriction fragment length polymorphism (RFLP) analysis of the phospholipase B gene (PLB1) as a simple tool to differentiate between C . neoformans subgroups . A PLB1 fragment, 1,970 bp, was amplified and digested with either AvaI or HindIII . Both sets of profiles grouped the isolates into their respective varieties, but only the AvaI profiles allowed for the identification of the eight molecular types via the corresponding RFLP profiles A1 to A8 . Digestion of the same fragments with HindIII resulted in RFLP profiles H1 to H5, which distinguished only between serotype A, AD, D, and B/C . Neither enzyme distinguished serotype B from serotype C . The serotype AD profile was a composite of the serotype A and D profiles . Further investigation showed that the serotype AD isolates used in this study are heterozygous, with one allele of PLB1 originating from a serotype A parent and the other from a serotype D parent. Eur J Immunol, 2003 Apr, 33(4), 1041 - 51 Differences in outcome of the interaction between Cryptococcus neoformans glucuronoxylomannan and human monocytes and neutrophils; Monari C et al.; Disseminated infections by the opportunistic yeast Cryptococcus neoformans are characterized by accumulation in tissues of glucuronoxylomannan (GXM), the major component of the capsular polysaccharide . We investigated binding, uptake, and disposal of GXM by peripheral blood neutrophils and monocytes, and the effect of GXM uptake on phagocytic cell function . GXM was efficiently bound and internalized by both types of phagocytic cells, with maximal loading at 50 microg/ml, a GXM concentration found in serum and cerebrospinal fluid of some cryptococcosis patients . However, substantial differences were noted in the kinetics for uptake by macrophages and neutrophils . Whereas neutrophils rapidly ingested limited amounts of GXM and then expelled or degraded it after 1 h of incubation, macrophages demonstrated continuous intracellular accumulation for up to 1 week of incubation . Accumulation of GXM by neutrophils was accompanied by reduced anticryptococcal activity, suggesting one more mechanism for virulence enhancement by the major capsular component of C . neoformans. Arkh Patol, 2003 Jan-Feb, 65(1), 42 - 5 {Pathological anatomy of HIV infection according to data in Saint-Petersburg}; Tsinzerling VA et al.; The results of postmortem investigation of 101 cases of HIV infection at its different stages in Saint-Petersburg are analyzed . The challenges to a pathologist performing the autopsy are discussed . There has been an increase in the number of deaths since 2000, among drug users in particular . Macrophacytic transformation and specific HIV-related encephalitis are among the most important changes directly associated with immunodeficiency virus . Among secondary infections, cryptococcosis, cytomegaly, pneumocystosis, and tuberculosis are most common and significant. Vet Clin Pathol, 1993, 22(4), 109 - 116 Chylothorax associated with cryptococcal mediastinal granuloma in a cat; Meadows RL et al.; A 10-year-old neutered female cat had chylothorax, precaval syndrome, and a mediastinal granuloma resulting from infection with Cryptococcus neoformans . Diagnosis of a chylous effusion was made by cytologic examination of pleural fluid and by finding higher triglyceride levels in the effusion than in serum (825 vs . 64 mg/dl, respectively) . Postmortem examination revealed cryptococcal organisms in the mediastinal granuloma, lungs, cerebral meninges, and connective tissues adjacent to the thyroid gland . Chylous effusion in a cat associated with cryptococcosis has not been reported previously . Cryptococcosis should be included in the differential diagnosis in chylous effusions in cats. J Antimicrob Chemother, 2003 May, 51(5), 1223 - 9 Epub 2003 Mar 28. In vitro and in vivo activity of tea tree oil against azole-susceptible and -resistant human pathogenic yeasts; Mondello F et al.; A tea tree oil (TTO) preparation of defined chemical composition was studied, using a microbroth method, for its in vitro activity against 115 isolates of Candida albicans, other Candida species and Cryptococcus neoformans . The fungal strains were from HIV-seropositive subjects, or from an established type collection, including reference and quality control strains . Fourteen strains of C . albicans resistant to fluconazole and/or itraconazole were also assessed . The same preparation was also tested in an experimental vaginal infection using fluconazole-itraconazole-susceptible or -resistant strains of C . albicans . TTO was shown to be active in vitro against all tested strains, with MICs ranging from 0.03% (for C . neoformans) to 0.25% (for some strains of C . albicans and other Candida species) . Fluconazole- and/or itraconazole-resistant C . albicans isolates had TTO MIC50s and MIC90s of 0.25% and 0.5%, respectively . TTO was highly efficacious in accelerating C . albicans clearance from experimentally infected rat vagina . Three post-challenge doses of TTO (5%) brought about resolution of infection regardless of whether the infecting C . albicans strain was susceptible or resistant to fluconazole . Overall, the use of a reliable animal model of infection has confirmed and extended our data on the therapeutic effectiveness of TTO against fungi, in particular against C . albicans. Mikrobiol Z, 2002 Nov-Dec, 64(6), 41 - 6 {Search for lectin-producers among representatives of some yeast genera}; Babich TV et al.; Hemagglutinating properties of the culture liquid and cells of 451 strains, 24 species, 7 genera of the museum and fresh yeast cultures have been investigated . The researchers have distinguished 333 strains (74%) capable for production of extracellular lectins and those bound to the cell surface with activity about 512-1024 hemagglutinating units . The extracellular lectins were found in 41% of lectin-synthesizing strains; the highest amount of their producers were found among representatives of genera Saccharomyces, Kluyveromyces, Cryptococcus . Strains of Debaryomyces genus mainly synthesized lectins of the bound form, and the yeast of Pichia genus--lectins of the both types . Affinity of the extracellular lectin Saccharomyces cerevisiae UCM Y-530 for D-gluconic acid has been established. Klin Oczna, 2002, 104(5-6), 421 - 3 {Ophthalmic complications in the course of opportunistic infections: part II}; Raczynska K et al.; In the second part of the paper we present ophthalmic complications of fungal infections in immunocompromised patients . Among about 200,000 known species of fungi, only approximately 200 are pathogenic for human . Many of them are not able to cause a disease in persons with normal immunity, but in immunocompromised patients they are the cause of severe, frequently lethal disorder . Most important agents are Candida albicans, Cryptococcus neoformans, Aspergillus fumigatus and Histoplasma capsulatum . Pneumocystis carnii, formerly considered to be a protozoan, currently is rated among fungi, and plays a very important role as a opportunistic pathogen in patients with AIDS . There are presented data about ocular involvement during fungal opportunistic infections--symptoms and clinical picture as well as methods of diagnosing and treatment. Oral Dis, 2003 Mar, 9(2), 55 - 61 Parotid gland involvement in advanced AIDS; Vargas PA et al.; OBJECTIVE: This study describes the involvement and the histological alterations found in the parotid glands of 100 patients who died with AIDS . MATERIALS AND METHODS: Sex, age, CD4 cell count and clinical history were obtained from the files of 100 patients who died with AIDS . Histological analysis of the parotid glands was performed using H&E, Gomori-Grocott, Ziehl-Neelsen and Mucicarmine . Histological findings were grouped in reactive, infectious, cystic, neoplastic and concomitant lesions . RESULTS: None of the patients presented complaints or symptoms related to salivary gland alterations prior to death . The mean age of the patients and CD4 cell count were 36.4 years and 76.07 cells microliter-1, respectively . Histological alterations of the parotid glands were found in 51% of the patients . The most common alteration was non-specific chronic sialadenitis (29 cases), followed by infectious conditions (22 cases) . Mycobacteriosis was the most common infectious disease (10 cases), followed by cytomegalovirus (nine cases), cryptococcosis (three cases) and histoplasmosis (two cases) . Lymphoepithelial cysts occurred in six cases, Warthin's tumor and non-Hodgkin Lymphoma in one case each . CONCLUSIONS: These results indicate that infection and other lesions in the parotid glands are more frequent than hitherto described in the specialized literature in AIDS patients . Clinicians should consider parotid gland involvement, when evaluating disease extension in advanced AIDS patients. Infect Immun, 2003 Apr, 71(4), 1988 - 94 Cryptococcus neoformans with a mutation in the tetratricopeptide repeat-containing gene, CCN1, causes subcutaneous lesions but fails to cause systemic infection; Chung S et al.; We studied a Cryptococcus neoformans strain that caused feline chronic nasal granuloma without disseminated disease . This strain, B-4551, grows at temperatures up to 35 degrees C and fails to cause systemic infection in mice . Many cells of B-4551 formed short hyphal elements in feline nasal tissue and occasionally at 35 degrees C in vitro . A complementation and sequence analysis revealed that the temperature-sensitive (Ts) phenotype of B-4551 was due to deletion of a lysine residue in the cryptococcal CCN1 gene . B-4551 complemented with the wild type CCN1 gene grew at 37 degrees C and caused fatal systemic infection in mice . The CCN1 gene encodes a protein containing 16 copies of a tetratricopeptide repeat . CCN1 is homologous to the Saccharomyces cerevisiae CLF1 gene, which is required for pre-mRNA splicing, cell cycle progression, and DNA replication, and to the Drosophila melanogaster crn gene, which is involved in neurogenesis . CLF1 complemented the Ts phenotype of B-4551 . CCN1, however, failed to rescue the clf1 mutant in S . cerevisiae . These results indicate that the Ccn1p may not be as functionally diverse as Clf1p in yeast. Anal Biochem, 2003 Mar 15, 314(2), 266 - 80 Site-specific characterization of the N-linked oligosaccharides of a murine immunoglobulin M by high-performance liquid chromatography/electrospray mass spectrometry; Wang F et al.; Immunoglobulin M is an especially important product of the immune system because it plays a critical role in early protection against infections . In this report, the glycosylation pattern of the protective murine monoclonal IgM 12A1 to Cryptococcus neoformans polysaccharide was analyzed by high-performance liquid chromatography coupled with electrospray ionization mass spectrometry . Peptide mapping studies covering 88% of the deduced amino acid sequence indicated that of the six potential N-glycosylation sites in this antibody only five were utilized, as the tryptic peptide derived from monoclonal IgM 12A1 containing Asn-260 was recovered without carbohydrates . The oligosaccharide side chains of monoclonal IgM 12A1 were characterized at each of the N-glycosylation sites . Asn-166 possessed 20 monosialylated and nonsialylated, and fucosylated and nonfucosylated complex- and hybrid-type oligosaccharides and one high-mannose-type oligosaccharide . Thirteen oligosaccharides were attached to the site at Asn-401, including six complex-type, four hybrid-type, and three high-mannose-type oligosaccharides . Twelve hybrid-type oligosaccharides were attached to Asn-378, three of which had terminal sialic acids . Eleven hybrid-type oligosaccharides were attached to Asn-331, seven of which had terminal sialic acids . Only two high-mannose type oligosaccharides were attached to Asn-363 . These results indicated great complexity in the structure and composition of oligosaccharides attached to individual IgM glycosylation sites. Tree Physiol, 2000 Apr, 20(7), 485 - 491 Genetic diversity and bark physiology of the European beech (Fagus sylvatica): a coevolutionary relationship with the beech scale (Cryptococcus fagisuga); Krabel D et al.; In 1994 and 1995, the degree of infestation by the beech scale (Cryptococcus fagisuga Lind.) was recorded on 120 beech trees (Fagus sylvatica L.) in the Pless Forest near Gottingen, Germany . Simultaneously, the trees were characterized genetically and compounds of primary and secondary metabolism of beech bark were analyzed . A correlation was established between beech scale infestation and the genotype of the host trees, based on gene locus A of isocitrate dehydrogenase (IDH-A) . The fraction of infested beech trees was higher in the heterozygous genotype A2A3 group than in the homozygous genotype A2A2 and A3A3 groups, whereas the fraction of beech trees with decreasing infestation from one year to the next was lower in the heterozygous genotype A2A3 group than in the homozygous genotype A2A2 and A3A3 groups . Concentrations of soluble carbohydrates, protein amino acids and proanthocyanidins were determined in the inner and outer bark of trees with differing degrees of infestation . The results indicate that the defense reaction of beech against infestation by beech scale comprises multiple processes in which nutrient availability to the beech scale is reduced by concentration shifts and by the formation of inhibiting compounds. Arch Soc Esp Oftalmol, 2003 Feb, 78(2), 103 - 6 {Cryptococcal choroiditis in a patient with acquired immunodeficiency syndrome}; Fernandez Gonzalez MC et al.; PURPOSE/METHODS: We report the case of a 31 year old male with presumed cryptococcal choroiditis associated with cryptococcal meningitis and AIDS . Multiple yellowish-whitish small choroidal lesions, retinal hemorrhages and cotton-wool spots appeared in the posterior pole associated with vision loss . We also carried out differential diagnosis with other frequent diseases and a review of the treatments . RESULTS/CONCLUSIONS: Cryptococcus neoformans is an opportunistic fungus with a predilection for infecting the meninges in patients with Acquired Immunodeficiency Syndrome AIDS (10%) . Ocular manifestation usually includes cranial nerve palsies or papilledema secondary to increased intracranial pressure . Cryptococcal choroiditis is less frequent (5% of patients with meningitis). J Immunol, 2003 Apr 1, 170(7), 3621 - 30 More is not necessarily better: prozone-like effects in passive immunization with IgG; Taborda CP et al.; Despite a century of study, the relationship between Ag-specific Ig concentration and protection remains poorly understood for the majority of pathogens . In certain conditions, administration of high Ab doses before challenge with an infectious agent can be less effective than smaller Ab doses, a phenomenon which is consistent with a prozone-like effect . In this study, the relationship between IgG1, IgG2a, IgG2b, and IgG3 dose, infective inocula, and protection was investigated in a mouse model of Cryptococcus neoformans infection . The activity of each IgG subclass ranged from protective to disease-enhancing depending on both the Ab dose and infective inocula used . Enhanced dissemination to the brain was observed in mice given a high IgG2a dose and a relatively low inoculum . Ab administration had immunomodulatory effects, with cytokine expression in lung, brain, and spleen varying as a function of the infective inoculum Ab dose and IgG subclass . In vitro studies did not predict or explain the mechanism of in vivo prozone-like effects, because all isotypes were opsonic and elicited NO release from macrophages . IgG2a was most efficient in inducing a macrophage oxidative burst . These results reveal that an individual Ab can be protective, nonprotective, or disease-enhancing depending on its concentration relative to a challenge inoculum . Our findings have implications for the potential contribution of Ab responses to defense against microbial diseases because Ab-mediated immunity may be protective, nonprotective, or even deleterious to the host. Indian J Med Sci, 2002 Jul, 56(7), 325 - 9 Prevalence of fungal meningitis among HIV positive & negative subjects in Indore (MP state); Jaiswal SP et al.; The aim of the study was to find out the prevalence of fungal meningitis among AIDS cases and to assess the prognosis of fungal meningitis among HIV positive & negative subjects . The study comprised of 15 & 10 cases of fungal meningitis among HIV positive & negative subjects respectively during the study period 1992-2001 . India ink preparation and Gram's staining procedures were carried out on the centrifuged CSF deposits . The CSF deposits were also used for bacterial and fungal culture . In the present study the prevalence of fungal meningitis was noted among 15 (3.1%) of 483 AIDS cases . Twelve of them had cryptococcal meningitis while 3 were infected with Candida albicans . Four AIDS cases presented fungal meningitis as a primary opportunistic infection and HIV status was confirmed in 4 of them after the diagnosis of fungal meningitis . 13 of the 15 cases were in the age group 26-40 yrs while one was 55 yrs old and the other 16 month old child; these two cases had blood transfusion transmitted and vertically transmitted mode of HIV transmission respectively . Further, only two of 15 cases were females and both acquired HIV infection through blood transfusion . Overall prognosis of fungal meningitis was poor among HIV positives and 7 of the 15 cases died in hospital within 2-3 weeks after diagnosis of cryptococcal meningitis . Among HIV negative subjects, ten cases of fungal meningitis (3 with Candida albicans and 7 with Cryptococcous neoformans) were noted at our end and nine of them had immunocompromised status (3 cases of renal transplant on immunosuppressives, 3 cases neonates/infant and 2 diabetic subjects . The fungal meningitis is one of the important causes of morbidity & mortality among immunocompromised among HIV positive subjects. AIDS Res Hum Retroviruses, 2003 Feb, 19(2), 85 - 90 Human immunodeficiency virus type 1 subtype and other factors associated with extrapulmonary Cryptococcosis among patients in Thailand with AIDS; Amornkul PN et al.; Delineating factors associated with extrapulmonary cryptococcosis (EPC), a major disease burden among Thailand's AIDS patients, can clarify its pathogenesis and guide preventive interventions . From November 1993 through June 1996, enhanced surveillance of 2261 human immunodeficiency virus (HIV)-seropositive patients in a hospital near Bangkok showed EPC among 561 of 1553 AIDS patients (36.1%) . Univariate analysis results were confirmed by multivariate analyses of data on 1259 patients . Logistic regression models identified factors significantly associated with EPC to be male sex (adjusted odds ratio {aOR}, 2.0; 95% confidence interval {CI}, 1.3-2.9), age <33 years (aOR, 1.5; 95% CI, 1.2-1.9), severe immunosuppression (aOR, 1.8; 95% CI, 1.3-2.6), not injecting drugs (aOR, 3.0; 95% CI, 1.7-5.5), and infection with HIV-1 circulating from CRF01_AE (formerly subtype E) versus subtype B (aOR, 2.3; 95% CI, 1.2-4.5) . The association with CRF01_AE may result from undetermined markers of exposure or viral subtype effects on host immune responses . Better understanding of the epidemiology of EPC may reduce EPC incidence through targeted primary prevention and treatment. Rev Med Chir Soc Med Nat Iasi, 2002 Jan-Mar, 106(1), 161 - 3 {Fungal infection in AIDS patients . Consideration for 10 cases}; Mihalache D et al.; Clinic, etiologic and therapeutic study of fungic infection in AIDS patients . The retrospective clinical study of the 10 AIDS patients admitted between 01.01.1994 and 31.12.2000 in Infectious Diseases Clinical Hospital of Iasi . The majority of the cases (7) were registered in the last 3 years: 4 children and 6 adults . HIV infection was known only for 4 patients according to their history, and for the other 6 patients this diagnosis was made concomitantly of fungic infection diagnosis . The disease onset was insidious in 7 cases (2 children and 5 adults) being characterized by manifestations of nervous system involvement and consciousness disturbances (6 cases) . The etiological diagnosis was established for alive patients by cerebrospinal fluid culture and hemocultures and Cryptococcus neoformans was isolated for 9 patients and Candida to another one . Fungic infection lead to meningeal injuries to 6 patients and the systemic one in for 4 patients . Although, under antifungical and antiretroviral therapy, the disease was lethal for 4 patients . The fungic infection recrudesces in AIDS patients, possessing systemic or/and meningeal manifestation, long evolution that can be lethal for a lot of them. Med Mycol, 2003 Feb, 41(1), 75 - 81 CAY-1, a novel antifungal compound from cayenne pepper; Renault S et al.; CAY-1, a novel saponin from Capsicum frutescens (commercially known as cayenne pepper) was investigated to determine its in vitro antifungal activity, mechanism of action and mammalian cell cytotoxicity . CAY-1 was active against 16 different fungal strains, including Candida spp . and Aspergillus fumigatus {minimum inhibitory concentrations (MIC) ranging from 4 to 16 microg ml(-1)}, and was especially active against Cryptococcus neoformans (90% inhibition at 1 microg ml(-1)) . Synergistic activity was also observed between CAY-1 and amphotericin B against Candida albicans and A . fumigatus . No significant cytotoxicity was demonstrated when CAY-1 was tested against 55 mammalian cell lines at up to 100 microg ml(-1) . Importantly, CAY-1 appears to act by disrupting the membrane integrity of fungal cells. Med Mycol, 2003 Feb, 41(1), 59 - 63 Pathogenicity of Cryptococcus neoformans var . gattii in an immunocompetent mouse model; Torres-Rodriguez JM et al.; The pathogenicity of two different genomic profiles of Cryptococcus neoformans var . gattii serotype B isolated from goats that died from cryptococcal pneumonia was assessed in an experimental model of immunocompetent mice . One strain of each randomly amplified polymorphic DNA (RAPD) profile (GR52 and GR56) and three reference C . neoformans isolates representing serotypes B, D and C were used . BALB/c male mice were inoculated by the intraperitoneal route with each strain . After 4 weeks of follow-up, the animals were sacrificed and autopsy specimens of testes, liver, spleen, kidney, lungs and brain were cultured and stained for histopathology . Although spontaneous mortality was only 2% (one animal), all mice except for those inoculated with serotype C showed positive cultures in almost one organ . The strain GR52 isolated from goat showed the highest rate of positive cultures (80%) followed by serotype D (77%) . Serotype B reference strain and second goat strain GR56 were both isolated from 70% of samples . Serotype C was recovered in only 33% of organs, and never from brain or lung specimens . GR52 grew abundantly from all lung cultures, and yeast cells with large capsules were seen in histopathology inside the alveoli, peribronchial vessels and interalveolar spaces . They appeared to elicit no inflammatory response . We conclude that intraperitoneally inoculated C . neoformans var . gattii shows high virulence in this immunocompetent mouse model . Strain GR52 was highest in pathogenicity and had marked lung tropism . In contrast, the serotype C reference strain showed the lowest pathogenicity and seemed not to spread outside the abdominal viscera. Clin Infect Dis, 2003 Mar 15, 36(6), 789 - 94 Epub 2003 Feb 27. The changing epidemiology of cryptococcosis: an update from population-based active surveillance in 2 large metropolitan areas, 1992-2000; Mirza SA et al.; To examine trends in the incidence and epidemiology of cryptococcosis, active, population-based surveillance was conducted during 1992-2000 in 2 areas of the United States (the Atlanta, Georgia, and Houston, Texas, metropolitan areas; combined population, 7.4 million) . A total of 1491 incident cases were detected, of which 1322 (89%) occurred in HIV-infected persons . The annual incidence of cryptococcosis per 1000 persons with AIDS decreased significantly during the study period, from 66 in 19