Mycoses, 2004 Apr, 47(3-4), 93 - 103 The role of complement in invasive fungal infections; Speth C et al.; New therapeutic approaches enable organ transplantations and guarantee longer survival for AIDS patients or patients with haematological neoplasia . The price for these medical advances is immunosuppression and thus enhanced susceptibility to opportunistic fungal infections . As a consequence invasive fungal infections are on the march in modern medicine . Therapeutic limitations and difficulties strongly demand for a deeper understanding of the interaction between the various fungi and the hosts' innate and adaptive immune defence system . This understanding is the essential prerequisite for a potential therapeutic approach, which may support specifically the insufficient antifungal attack of the host . In the present article, we therefore review the current knowledge of the role of the complement system as a central part of innate immunity and as a fine tuner of adaptive immunity in the pathogenesis of invasive fungal infections, such as aspergillosis, candidosis, cryptococcosis, paracoccidioidomycosis, blastomycosis and histoplasmosis. Eukaryot Cell, 2004 Apr, 3(2), 413 - 9 Challenge of Drosophila melanogaster with Cryptococcus neoformans and role of the innate immune response; Apidianakis Y et al.; We found that the ingestion of Cryptococcus neoformans by Drosophila melanogaster resulted in the death of the fly but that the ingestion of Saccharomyces cerevisiae or the nonpathogenic Cryptococcus kuetzingii or Cryptococcus laurentii did not . The C . neoformans protein kinase A and RAS signal transduction pathways, previously shown to be involved in virulence in mammals, also played a role in killing DROSOPHILA: Mutation of the Toll immune response pathway, the predominant antifungal pathway of the fly, did not play a role in Drosophila defense following ingestion of the yeast . However, the Toll pathway was necessary for the clearance of C . neoformans introduced directly into the hemolymph of D . melanogaster and for the survival of systemically infected flies. Eukaryot Cell, 2004 Apr, 3(2), 385 - 92 Cryptococcus neoformans CAP59 (or Cap59p) is involved in the extracellular trafficking of capsular glucuronoxylomannan; Garcia-Rivera J et al.; Several genes are essential for Cryptococcus neoformans capsule synthesis, but their functions are unknown . We examined the localization of glucuronoxylomannan (GXM) in strain B-3501 and in cap59 mutants B-4131 and C536 . Wild-type strain B-3501 showed a visible capsule by India ink staining and immunofluorescence with anticapsular monoclonal antibodies (MAbs) 12A1 and 18B7 . B-4131, a mutant containing a missense mutation in CAP59, showed no capsule by India ink staining but revealed the presence of capsular polysaccharide on the cell surface by immunofluorescence . The cap59 gene deletion mutant (C536), however, did not show a capsule by either India ink staining or immunofluorescence . Analysis of cell lysates for GXM by enzyme-linked immunosorbent assay revealed GXM in C536 samples . Furthermore, the epitopes recognized by MAbs 12A1, 2D10, 13F1, and 18B7 were each detected in the cytoplasm of all strains by immunogold electron microscopy, although there were differences in location consistent with differences in epitope synthesis and/or transport . In addition, the cells of B-3501 and B-4131, but not those of the cap59 deletant, assimilated raffinose or urea . Hence, the missense mutation of CAP59 in B-4131 partially hampered the trafficking of GXM but allowed the secretion of enzymes involved in hydrolysis of raffinose or urea . Furthermore, the cell diameter and volume for strain C536 are higher than those for strain B-3501 or B-4131 and may suggest the accumulation of cellular material in the cytoplasm . Our results suggest that CAP59 is involved in capsule synthesis by participating in the process of GXM (polysaccharide) export. Mikrobiologiia, 2004 Jan-Feb, 73(1), 111 - 7 {Groups and sources of yeasts in house dust}; Glushakova AM et al.; House dust contains bacteria, mycelial fungi, microarthropods, and yeasts . The house dust samples collected in 25 apartments in Moscow and the Moscow region were found to contain yeasts belonging to the genera Candida, Cryptococcus, Debaryomyces, Rhodotorula, Sporobolomyces, and Trichosporon . The most frequently encountered microorganisms were typical epiphytic yeasts, such as Cryptococcus diffluens and Rhodotorula mucilaginosa, which are capable of long-term preservation in an inactive state . The direct source of epiphytic yeasts occurring in the house dust might be the indoor plants, which were contaminated with these yeasts, albeit to a lesser degree than outdoor plants . Along with the typical epiphytic yeasts, the house dust contained the opportunistic yeast pathogens Candida catenulata, C . guillermondii, C . haemulonii, C . rugosa, and C . tropicalis, which are known as the causal agents of candidiasis . We failed to reveal any correlation between the abundance of particular yeast species in the house dust, residential characteristics, and the atopic dermatitis of the inhabitants. Microbiology, 2004 Apr, 150(Pt 4), 785 - 93 Coronamycins, peptide antibiotics produced by a verticillate Streptomyces sp . (MSU-2110) endophytic on Monstera sp; Ezra D et al.; Coronamycin is a complex of novel peptide antibiotics with activity against pythiaceous fungi and the human fungal pathogen Cryptococcus neoformans . It is also active against the malarial parasite, Plasmodium falciparum, with an IC(50) of 9.0 ng ml(-1) . Coronamycin is produced by a verticillate Streptomyces sp . isolated as an endophyte from an epiphytic vine, Monstera sp., found in the Manu region of the upper Amazon of Peru . Bioassay-guided fractionation of the fermentation broths of this endophyte on silica gel and HPLC chromatography yielded two principal, inseparable, peptides with masses of 1217.9 and 1203.8 Da . Three other minor, but related components, are also present in the preparation . Amino acid analysis of coronamycin revealed residues of component 1, component 2, methionine, tyrosine and leucine at a ratio of 2:2:1:1:3 . Other compounds with antifungal activities are also produced by this endophytic streptomycete. J Antimicrob Chemother, 2004 May, 53(5), 750 - 8 Epub 2004 Apr 08. In vitro synergic antifungal effect of MUC7 12-mer with histatin-5 12-mer or miconazole; Wei GX et al.; OBJECTIVES: MUC7 12-mer (RKSYKCLHKRCR), a cationic peptide derived from human salivary MUC7 mucin, exhibits potent in vitro antifungal activity, as determined by killing assays in phosphate buffer . In this study we examined the MUC7 12-mer antifungal activity alone or in combination with other antifungal agents in LYM medium (modified RPMI 1640) . METHODS: Antifungal activities of MUC7 12-mer and other compounds against several fungal strains were first measured by MIC and minimum fungicidal concentration (MFC) tests using broth microdilution assay . The viability of Candida albicans and Cryptococcus neoformans were also determined by killing assays and time kinetics of peptide-mediated killing . Antifungal activities of MUC7 12-mer in combination with other compounds {histatin-5 (Hsn5) 12-mer: AKRHHGYKRKFH, amphotericin B or miconazole} against C . albicans and C . neoformans were determined by chequerboard assays and confirmed by killing assays . Toxicities of individual compounds were determined by haemolytic assays . RESULTS: MICs and MFCs of MUC7 12-mer ranged from 3.13 to 6.25 mg/L for most of the strains tested, and were, in most cases, comparable to those of amphotericin B and miconazole (0.78-6.25 mg/L) . ED(50) values of MUC7 12-mer and Hsn5 12-mer were 7.1 and 7.4 micro M (or 11.2 and 11.6 mg/L), respectively, for C . albicans; and 1.2 and 1.1 micro M (or 1.9 and 1.7 mg/L), respectively, for C . neoformans . The killing of C . albicans and C . neoformans was achieved after 30 and 10 min exposure to the peptides, respectively . Combinations of MUC7 12-mer and Hsn5 12-mer, and of MUC7 12-mer and miconazole have a synergic antifungal effect on C . neoformans, with a fractional inhibitory concentration index (FICI) of 0.37 and 0.25, respectively; and a slightly lower than synergic effect on C . albicans, with a FICI of 0.63 and 0.56, respectively . In addition, using human erythrocytes, the two salivary peptides showed low levels of haemolytic activity . CONCLUSIONS: This study suggests that MUC7 12-mer and Hsn5 12-mer peptides may be suitable candidates for use in combination antifungal therapy. Drug Resist Updat, 2004 Feb, 7(1), 3 - 10 Immunotherapy for invasive fungal infections: from bench to bedside; Pappas PG; Immunotherapy for invasive fungal infections has been an area of significant research and clinical interest since the first-half of the twentieth century when hyperimmune equine immunoglobulin was first successfully administered to patients with acute and chronic cryptococcal meningitis . Since that time, effective antifungal compounds have been developed, but a much more complex array of host disorders have also emerged creating an even greater need for immunotherapy in conjunction with conventional antifungal therapy . In this review, the scientific foundation supporting the use of various immunotherapeutic interventions including granulocyte infusions, cytokine growth factors, pro-inflammatory cytokines including IFN-gamma and interleukin-12, immunoglobulin therapy, and active immunization will be discussed . Clinical data supporting the use of these interventions are often scant and inconclusive, however, relevant clinical information will be presented . In theory, adjunctive immunotherapy for invasive fungal infections has significant potential for improving clinical outcomes in a growing population of patients at risk for these potentially devastating infections . Clearly, randomized double-blind clinical trials will need to be performed to better understand the precise role of these interventions . There are several obstacles preventing the conduct of these studies, but these pressing clinical issues must be addressed through carefully considered study design and effective implementation. Nihon Kokyuki Gakkai Zasshi, 2004 Mar, 42(3), 272 - 6 {A case of secondary pulmonary cryptococcosis complicating diabetes mellitus}; Tanigawa M et al.; In this case, a 65-year-old man complained of fever and productive cough while being treated for diabetes mellitus at the outpatient clinic . His chest CT scan revealed multiple infiltrative lesions in both the right and left lower pulmonary lobes . He was therefore given an antibiotic on suspicion of having bacterial pneumonia, and he also received nutritional instruction in relation to diabetes mellitus, and remission resulted . However, he could not maintain sufficient glycemic control thereafter, and his pulmonary lesions persisted . Because his lesions changed into cavitied multiple nodular lesions, as seen on a chest CT scan, a transbronchial lung biopsy was performed . Histopathological examination of the biopsy specimen demonstrated Cryptococcus organisms, and the Cryptococcus antigen titer was high, which led to a diagnosis of pulmonary cryptococcosis . After oral treatment with fluconazole for 1 year and 4 months, only a small nodule in the right lower lobe and a funicular lesion in the left lower lobe remained on a chest CT scan, and the patient had neither subjective symptoms nor evidence of inflammation, although he still had a positive antigen titer for Cryptococcus . Thus, the treatment was terminated . Improvement of the clinical symptoms and of the laboratory and radiological findings demonstrated the therapeutic efficacy of this treatment. Carbohydr Res, 2004 Apr 28, 339(6), 1047 - 60 Hydrolysis of Nothogenia erinacea xylan by xylanases from families 10 and 11; Nerinckx W et al.; The structures of several enzymatic hydrolysis products of Nothogenia erinacea seaweed xylan, a linear homopolymer with mixed beta-(1-->3)/beta-(1-->4) linkages, were analysed by physicochemical and biochemical techniques . With the glycoside hydrolase family 10 beta-(1-->4)-xylanase from Cryptococcus adeliae, hydrolysis proceeds to a final mixture of products containing a mixed linkage-type triose as a major compound, whereas with the family 11 xylanase from Thermomyces lanuginosus this is a mixed linkage tetraose . The Cryptococcus xylanase is shown to be capable of also catalysing the hydrolysis of beta-(1-->3) linkages, that is this of a mixed type tetraose intermediary formed, in accordance with the broader substrate specificity of family 10 enzymes . From a partial degradation experiment with the T . lanuginosus xylanase, a series of higher mixed oligosaccharides were isolated and identified . The observed oligosaccharide intermediates and splicing pattern indicate an irregular beta-(1-->3)/beta-(1-->4) linkage distribution within the linear d-xylose polymer . Similar results were obtained with rhodymenan, the seaweed xylan from Palmares palmata. J Med Assoc Thai, 2004 Feb, 87(2), 173 - 9 HIV infection/acquired immunodeficiency syndrome at Siriraj Hospital, 2002: time for secondary prevention; Anekthananon T et al.; The authors retrospectively reviewed the medical records of HIV/AIDS patients who were admitted to the medical service, Siriraj Hospital from January 1, 2002 through December 31, 2002 . Demographics, CD4 lymphocyte counts, discharge diagnoses, the incidence of Pneumocystis carinii pneumonia (PCP), cerebral toxoplasmosis and cryptococcosis in patients who received and did not receive appropriate chemoprophylaxis against those opportunistic infections when indicated, and outcome of the patients were collected . Three hundred medical records of 286 HIV/AIDS patients were available for review . One hundred and seventy two patients (60.1%) were male . Mean age of the patients was 36.8 +/- 9.91 years (range 14-74) . The mean CD4 lymphocyte count that was determined in 165 patients was 74.7 +/- 134.21 cells/mm3 (range 0-894) . Of the 300 admissions, 36 per cent were newly diagnosed HIV infection . Only 23 (7.7%) patients had received antiretroviral drugs at the time of hospitalization . The leading HIV-related diseases were tuberculosis (29.3%), Pneumocystis carinii pneumonia (18.7%), and cryptococcosis (15.7%) . The rest of them included cytomegalovirus diseases (6.3%), lymphoma (6.3%), Salmonella bacteremia (6%), cerebral toxoplasmosis (5.7%), cryptosporidiosis (5.3%), disseminated Mycobacterium avium complex infection (1.0%), extrapulmonary histoplasmosis (1.0%), Candida esophagitis (1.0%), progressive multifocal leukoencephalopathy (1.0%), and rhodococcosis (0.7%) . Among those for whom HIV infection was established and chemoprophylaxis for PCP, cerebral toxoplasmosis and cryptococcosis were indicated, 9.8 per cent vs 28.2 per cent, 3.6 per cent vs 5.1 per cent, and 10 per cent vs 15.2 per cent of whom received and did not receive the appropriate chemoprophylaxis developed PCP, cerebral toxoplasmosis and cryptococcosis respectively . One hundred and ninety (63.3%) patients were alive at discharge, 84 (28.0%) had died, 21 (7%) were referred to other hospitals, and 5 (1.7%) left hospital against medical advice . The mortality rate in newly diagnosed HIV and in known HIV without antiretroviral treatment were comparable but much lower in known HIV-infected patients who received antiretroviral therapy . Secondary prevention by detection of HIV-infected patients while they are asymptomatic and providing them with appropriate chemoprophylaxis against specific opportunistic infections as well as appropriate antiretroviral treatment would decrease morbidity, mortality, and improve the quality of life of HIV-infected patients in Thailand. Lett Appl Microbiol, 2004, 38(5), 383 - 7 Isolation and characterization of psychrophilic yeasts producing cold-adapted pectinolytic enzymes; Nakagawa T et al.; AIMS: The present study was conducted to screen for psychrophilic yeasts that are able to degrade pectin compounds at low temperature, and to examine the cold-active pectinolytic enzymes produced by the isolated psychrophilic yeasts . METHODS AND RESULTS: Psychrophilic yeasts, which grow on pectin as a sole carbon source, pectinolytic-psychrophilic yeast (PPY) strains PPY-3, 4, 5 and 6, were isolated from soil from Abashiri (Hokkaido, Japan) . The sequences of 28S rDNA D1/D2 of strains PPY-3 and 4 indicated a taxonomic affiliation to Cryptococcus cylindricus and Mrakia frigida, respectively, strains PPY-5 and 6 belonged to Cystofilobasidium capitatum . The isolated strains were able to grow on pectin at below 5 degrees C, and showed the activities of several cold-active pectinolytic enzymes . CONCLUSION: The findings of this study indicate the possibility that the isolated strains produce novel pectinolytic enzymes that are able to degrade pectin compounds at low temperature . Significance and Impact of the Study: It is possible that the cold-active pectinolytic enzymes from the isolated strains can be applied to the food industry, e.g . the clarification of fruit juice below 5 degrees C. Mycoses, 2003, 46 Suppl 2, 15 - 20 {New therapeutical options for pulmonary fungal infections in non-neutropenic ICU-patients}; Hohl R; Pulmonary fungal infections in non-neutropenic ICU-patients are not frequent, however, their incidence is increasing with high morbidity and mortality, leading to prolonged stay in ICU wards and to excessive costs dependent on difficult delayed diagnosis . Candida as well as Aspergillus spp . are most important pathogens, but also species of less frequent genera must be taken into account, such as Fusarium, Scedosporium, Cryptococcus and others . Newly evaluated antimycotic agents, such as voriconazole and caspofungin, apart from fluconazole, are not only new options, but must be regarded as agents of choice in non-neutropenic patients. Arkh Patol, 2004 Jan-Feb, 66(1), 35 - 7 {Generalized cryptococcosis in HIV infection}; Parkhomenko IuG et al.; Two cases of generalized cryptococcosis in patients who died of HIV infection are described . The course of the disease was masked by other diseases and final diagnosis was established after necropsy . Leukemia was a clinical "mask" in one case and generalized tuberculosis in the other . Numerous cryptococci were found in different organs histologically with positive Shiff-reaction . Ultrastructurally, cryptococci were of variable forms this probably reflecting different stages of interaction between microorganisms and the host. Curr HIV Res, 2004 Jan, 2(1), 61 - 78 Advances in neuroimaging for HIV-1 associated neurological dysfunction: clues to the diagnosis, pathogenesis and therapeutic monitoring; Boska MD et al.; Persons with advanced human immunodeficiency virus type one (HIV-1) infection seek medical advice for a wide range of neurological disorders including, but not limited to, peripheral neuropathy, toxoplasmosis, cryptococcal meningitis, cytomegalovirus retinitis progressive multifocal leukoencephalopathy, lymphoma and dementia . The diagnosis of HIV-1-associated dementia (HAD) induced as a direct consequence of HIV infection of the brain comes commonly by exclusion . Diagnostic decisions can often be clouded by concomitant depression, motor impairments, and lethargy that follow debilitating immune suppression and weight loss . Indeed, cognitive, motor and behavior abnormalities underlie a variety of neurological dysfunctions associated with advanced HIV-1 infection . Thus, even combinations of clinical, laboratory and neuroimaging tests {for example, magnetic resonance imaging (MRI), computed tomography (CT), single photon emission computed tomography (SPECT) and positron emission tomography (PET)} often fail to provide conclusive diagnostic information . Nonetheless, the recent development of quantitative MR spectroscopic imaging has improved diagnostic possibilities for HAD . We are pleased to discuss these developments as well as taking a forward look into what will soon be made available to improve neuroimaging diagnostic precision . New MR and SPECT testing are being developed in our laboratories and elsewhere both for animal model systems and in humans with HIV-1 disease . Such tests can facilitate dynamic measures of HIV-1 neuropathogenesis providing information for disease events that even 2 years ago were unattainable. Korean J Intern Med, 2004 Mar, 19(1), 53 - 7 First report of Cryptococcus albidus--induced disseminated cryptococcosis in a renal transplant recipient; Lee YA et al.; Cryptococcus albidus, a non-neoformans species of the genus Cryptococcus, is generally regarded as a rare cause of disease . There have been only 14 previously reported cases in which this organism has been isolated as a pathogen, none of which occurred in a renal transplant recipient . A 23-year-old renal transplant recipient taking medication consisting of cyclosporine and prednisolone was admitted with a 10-day history of dry cough, fever and progressive dyspnea . The next day, his respiratory status deteriorated dramatically, and he developed acute respiratory distress syndrome (ARDS) and fulminant septic shock . On the eighth hospital day, tender macules on both his shins coalesced to form erythematous patches . Cryptococcus albidus was isolated by skin biopsy and tissue culture . We report here the first case of disseminated cryptococcosis caused by C . albidus in a renal transplant recipient who had been successfully treated with fluconazole monotherapy. Mol Microbiol, 2004 Apr, 52(1), 13 - 24 Molecular architecture of the Cryptococcus neoformans capsule; Gates MA et al.; Many microbes are surrounded by phagocytosis-inhibiting capsules . We took advantage of the large size of the polysaccharide capsule of the pathogenic yeast Cryptococcus neoformans to examine capsular architecture and the relationship between molecular architecture and the interaction of the capsule with potentially opsonic serum proteins . Our experimental design used complementary approaches in which (i) assessment of permeability to macromolecules of different Stokes radii; (ii) determination of the binding of Fab fragments of anticapsular antibodies as a measure of matrix density; (iii) capsular deconstruction by treatment with dimethyl sulphoxide; and (iv) evaluation of capsule plasticity, were used to probe the molecular structure of the capsule . The results showed that the capsule is a matrix with a variable porosity that increases with distance from the cell wall . A high density of the matrix at the capsule interior prevents penetration of large macromolecules to sites near the cell wall . In contrast, the capsular edge that is the interface with phagocytes presents capsular polysaccharide in a very low density that exhibits considerable plasticity and permeability to macromolecules . Notably, the capsule of yeast cells harvested from infected tissue showed a greater matrix density than yeast cells grown in vitro under capsule induction conditions. An Acad Bras Cienc, 2004 Mar, 76(1), 67 - 84 Epub 2004 Mar 04. Structure and biological functions of fungal cerebrosides; Barreto-Bergter E et al.; Ceramide monohexosides (CMHs, cerebrosides) are glycosphingolipids composed of a hydrophobic ceramide linked to one sugar unit . In fungal cells, CMHs are very conserved molecules consisting of a ceramide moiety containing 9-methyl-4,8-sphingadienine in amidic linkage to 2-hydroxyoctadecanoic or 2-hydroxyhexadecanoic acids, and a carbohydrate portion consisting of one residue of glucose or galactose . 9-Methyl 4,8-sphingadienine-containing ceramides are usually glycosylated to form fungal cerebrosides, but the recent description of a ceramide dihexoside (CDH) presenting phytosphingosine in Magnaporthe grisea suggests the existence of alternative pathways of ceramide glycosylation in fungal cells . Along with their unique structural characteristics, fungal CMHs have a peculiar subcellular distribution and striking biological properties . In Pseudallescheria boydii, Candida albicans, Cryptococcus neoformans, Aspergillus nidulans, A . fumigatus, and Schizophyllum commune, CMHs are apparently involved in morphological transitions and fungal growth . The elucidation of structural and functional aspects of fungal cerebrosides may therefore contribute to the design of new antifungal agents inhibiting growth and differentiation of pathogenic species. Sante, 2003 Oct-Dec, 13(4), 225 - 9 {Cryptococosis and HIV/AIDS: a review of 21 cases reported in the Tropical Diseases Centre, Ho Chi Minh City (Vietnam)}; Huynh TX et al.; Cryptococcus neoformans affections during HIV-infection are frequent and serious . The aim of this study was to analyse the epidemiological, clinical, biological and therapeutic characteristics of cryptococcal meningitis in HIV-positive patients, admitted into the Center for Tropical Diseases Ho Chi Minh City (Vietnam), during a 5-month period (May-September 2001) . Twenty-one patients (20 men and one woman) were included (identification of C . neoformans from the cerebrospinal fluid) . The mean age was 28 years . The majority of patients had been living in Ho Chi Minh City (48%) . The use of drugs and unprotected sexual relations were the principal risk factors of HIV-infection . The paucity and the confusion of clinical signs and symptoms, along with a high frequency of meningitis have been analysed . Clinical presentation features included: headache (95%), emaciation (90%), oro-pharyngeal candidiasis (90%), stiff neck (80%), nausea/vomiting (70%), fever (67%), coughing (38%), diarrhoea (33%), skin lesions (5%), convulsion (5%), photophobia (5%), and hemiparesis (5%) . The severity of the prognosis was mainly linked to the delay before hospitalization, to the possible association with other opportunistic infections, and to the availability of appropriate treatment. Curr Opin Investig Drugs, 2004 Feb, 5(2), 198 - 201 Amphotericin B cochleates: a vehicle for oral delivery; Perlin DS; Cochleates are a novel lipid-based delivery vehicle consisting of crystalline phospholipid-cation structures that form spiral lipid sheets . They represent a new technology platform for oral delivery of clinically important drugs that possess poor oral bioavailability . Orally administered cochleates containing amphotericin B (CAMB) showed broad-spectrum activity in murine infection models of candidiasis, aspergillosis and cryptococcosis . Initial biodistribution studies of CAMB administered orally in mice demonstrated that cochleates delivered significant levels of AMB to target organs . The lipid particulate nature of cochleates also imparted reduced toxicity that mimics other lipid-amphotericin B complexes . Cochleates are a promising new vehicle for oral delivery of amphotericin B at therapeutic levels. Rev Soc Bras Med Trop, 2004 Jan-Feb, 37(1), 65 - 6 Epub 2004 Mar 19. {Cryptococcal mastitis after corticosteroid therapy}; Ramos-Barbosa S et al.; We described a case of cryptococcal mastitis in a 46 years old female patient receiving corticosteroids for sarcoidosis . There was radiological pulmonary deterioration and a cystic lesion was found in the left breast.The mycosis was diagnosed by histopathology and cryptococcal antigens . We emphasized the sarcoid granuloma diagnostic's dilemma an the rare manifestation of cryptococcal infection as mastitis. FEMS Yeast Res, 2004 Mar, 4(6), 597 - 603 Diversity of Cryptococcus and Dioszegia yeasts (Basidiomycota) inhabiting arbuscular mycorrhizal roots or spores; Renker C et al.; The genera Cryptococcus and Dioszegia contain basidiomycetous yeasts found in a wide range of habitats . Primers to amplify the internal transcribed spacer (ITS) region of the nuclear ribosomal DNA (nrDNA) of arbuscular mycorrhizal fungi (AMF) also allow detecting members of this yeast group . Here we report the results of a sequence analysis using maximum parsimony on a set of 50 ITS sequences of yeasts associated with AMF structures (roots of 26 plant species, AM spores) from six field sites in Central Germany . Among 10 separated taxa, respectively five in the Tremellales and two in the Filobasidiales had unknown sequences . Therefore it was not possible to assign these sequences to any known species . The study indicates that exploring the diversity of Cryptococcus and Dioszegia in soil habitats with molecular methods might enlarge the actually estimated biodiversity of the group. Infect Immun, 2004 Apr, 72(4), 2338 - 49 Cytokine and inducible nitric oxide synthase mRNA expression during experimental murine cryptococcal meningoencephalitis; Maffei CM et al.; The immune events that take place in the central nervous system (CNS) during cryptococcal infection are incompletely understood . We used competitive reverse transcription-PCR to delineate the time course of the local expression of mRNAs encoding a variety of cytokines and inducible nitric oxide synthase (iNOS) during progressive murine cryptococcal meningoencephalitis and assessed the CNS inflammatory response using immunohistochemistry . Interleukin 18 (IL-18), transforming growth factor beta1, and IL-12p(40) mRNAs were constitutively expressed in the brains of infected and uninfected mice; IL-2 mRNA was not detected at any time . Increased levels of transcripts corresponding to IL-1 alpha, tumor necrosis factor alpha (TNF-alpha), and iNOS were detected as early as day 1 postinfection, with TNF-alpha rising by approximately 30-fold and iNOS increasing by approximately 5-fold by day 7 . Each remained at these levels thereafter . IL-4, IL-6, and gamma interferon transcripts were detected on day 5, and IL-1 beta and IL-10 transcripts were detected beginning on day 7 . Once detected, each remained at a relatively constant level through 28 days of infection . This cytokine profile does not suggest a polarized Th1 or Th2 response . Immunohistochemistry did not reveal inflammatory infiltrates before day 7, despite the presence of cryptococci . Intraparenchymal abscesses with inflammatory cells in their peripheries were found beginning on day 10 . The infiltrates were comprised primarily of cells expressing CD4, CD8, or CD11b; low numbers of cells expressing CD45R/B220 were also present . The persistence of Cryptococcus observed in the CNS may result from an ineffective immune response, perhaps owing to an insufficient anticryptococcal effector function of endogenous glial cells resulting from competing pro- and anti-inflammatory cytokines . These data detail the immune response in the brain and could be important for the future design of specific immunomodulatory therapies for this important opportunistic infection. Infect Immun, 2004 Apr, 72(4), 2229 - 39 Role of extracellular phospholipases and mononuclear phagocytes in dissemination of cryptococcosis in a murine model; Santangelo R et al.; Secreted phospholipase B (PLB) activity promotes the survival and replication of Cryptococcus neoformans in macrophages in vitro . We therefore investigated the role of mononuclear phagocytes and cryptococcal PLB in the dissemination of infection in a mouse model, using C . neoformans var . grubii wild-type strain H99, a PLB1 deletion mutant (Delta plb1), and a reconstituted strain (Delta plb1(rec)) . PLB facilitated the entry of endotracheally administered cryptococci into lung IM . PLB was also required for lymphatic spread from the lung to regional lymph nodes and for entry into the blood . Langhans-type giant cells containing budding cryptococci were seen free in the lymphatic sinuses of hilar nodes of H99- and Delta plb1(rec)-infected mice, suggesting that they may have a role in the dissemination of cryptococcal infection . The transfer of infected lung macrophages to recipient mice by tail vein injections demonstrated that these cells can facilitate hematogenous dissemination of cryptococci to the brain, independent of cryptococcal PLB secretion . PLB activities of cryptococci isolated from lung macrophages or infected brains were not persistently increased . We conclude that mononuclear phagocytes are a vehicle for cryptococcal dissemination and that PLB activity is necessary for the initiation of interstitial pulmonary infections and for dissemination from the lung via the lymphatics and blood . PLB is not, however, essential for the establishment of neurological infections when cryptococci are presented within, or after passage through, mononuclear phagocytes. Arch Pharm (Weinheim), 2004 Mar, 337(3), 152 - 5 Total synthesis and in vitro-antifungal activity of (+/-)-2-methoxytetradecanoic Acid; Carballeira NM et al.; The marine fatty acid (+/-)-2-methoxytetradecanoic acid was synthesized in two steps (71% overall yield) starting from commercially available methyl-2-hydroxy-tetradecanoate . The title compound was antifungal against Candida albicans (ATCC 14053) in RPMI medium and Aspergillus niger (ATCC 16404) and Cryptococcus neoformans (ATCC 66031) in SDB medium at the minimum inhibitory concentration (MIC) of 100 mM, which compares to the fungitoxicity of a 2-iodotetradecanoic acid against the same fungi . The title compound was also five to ten times more cytotoxic than capric acid to C . albicans and A . niger in the tested medium but comparable in cytotoxicity to either capric acid and its 2-methoxylated analog to C . neoformans . The antifungal activity of (+/-)-2-methoxytetradecanoic acid is explained in terms of inhibition of N-myristoyltransferase. J Ethnopharmacol, 2004 Mar, 91(1), 105 - 8 Antibacterial and antifungal activity of Syzygium jambolanum seeds; Chandrasekaran M et al.; The water and methanolic extracts of Syzygium jambolanum seeds were examined for antibacterial and antifungal activity in vitro using the disc diffusion method, minimum inhibitory concentration, minimum bactericidal concentration and minimum fungicidal concentration . Activity against gram positive bacteria (Bacillus subtilis, Staphylococcus aureus), gram negative bacteria (Salmonella typhimurium, Pseudomonas aeruginosa, Klebsiella pneumoniae and Escherichia coli) and fungal strains (Candida albicans, Cryptococcus neoformans, Aspergillus flavus, Aspergillus fumigatus, Aspergillus niger, Rhizopus sp., Trichophyton rubrum, Trichophyton mentagrophytes and Microsporum gypseum) is discussed. Antonie Van Leeuwenhoek, 2004 Jan, 85(1), 37 - 44 Induced synchrony in Cryptococcus neoformans after release from G2-arrest; Ohkusu M et al.; Cryptococcus neoformans was grown first to OD 4 under moderate aeration, then diluted 2.5 times with fresh medium, and grown under limited aeration for 5 h . Oxygen concentration decreased from 5-6 mg l(-1) to 1.5 mg l(-1) 1 h after the shift to limited aeration, and remained at a similar level thereafter . In all the eleven strains examined the shift caused unbudded G(2)-arrest in more than half of the cells . In three strains more than 80% of the cells were arrested in unbudded G(2), and, therefore they were selected for synchrony experiments . After being shifted to extensive aeration again, the cells resumed growth by synchronous budding, followed by synchronous nuclear division . This method has turned out to be a good tool to prepare synchronized culture in C . neoformans, especially when a large amount of synchronized cells is needed . This is worthy of attention, since synchronous cultures after release from G(2)-arrest have not been reported yet in any yeast species. Biochem J, 2004 Jul 1, 381(Pt 1), 131 - 6 Biosynthesis of UDP-GlcA, a key metabolite for capsular polysaccharide synthesis in the pathogenic fungus Cryptococcus neoformans; Bar-Peled M et al.; UDP-glucose dehydrogenase catalyses the conversion of UDP-glucose into UDP-GlcA, a critical precursor for glycan synthesis across evolution . We have cloned the gene encoding this important enzyme from the opportunistic pathogen Cryptococcus neoformans . In this fungus, UDP-GlcA is required for the synthesis of capsule polysaccharides, which in turn are essential for virulence . The gene was expressed in Escherichia coli and the 51.3-kDa recombinant protein from wild-type and five mutants was purified for analysis . The cryptococcal enzyme is strongly inhibited by UDP-xylose and NADH, has highest activity at pH 7.5 and demonstrates Km (app) values of 0.1 and 1.5 mM for NAD+ and UDP-glucose respectively . Its activity was significantly decreased by mutations in the putative sites of NAD+ and UDP-glucose binding . Unlike previously reported eukaryotic UDP-glucose dehydrogenases, which are hexamers, the cryptococcal enzyme is a dimer. Indian J Pathol Microbiol, 2003 Jul, 46(3), 416 - 9 Needle necropsy in AIDS; Satyanarayana S et al.; The value of autopsy in understanding the natural course of any disease is beyond any argument . The reluctance of pathologists to perform autopsy in HIV infected cadavers is justified due to the risks involved to the prosector and the morgue attendants . A relative low risk needle necropsy protocol is proposed using fine needle aspiration cytology, tru-cut biopsies and microbiological examination . Diagnosis could be offered in all the forty-four needle necropsies performed . Disseminated tuberculosis in 18/44 (40.9%) cases, disseminated cryptococcosis in 12/44 (27.2%) cases, poly-microbial infections in 27.2% cases and non-Hodgkin's lymphoma in 9% cases were detected in the study . Infectious agents like Histoplasma capsulatum, Isospora belli, tachyzoites of Toxoplasma gondii, Candida sp and Cryptococcus sp could be demonstrated in the samples obtained in the study . Lack of material for study of gross pathology, inaccessibility of deep-seated lesions and risk of needle stick injury to the prosector though low are the limitations of this procedure. Diagn Microbiol Infect Dis, 2004 Mar, 48(3), 201 - 5 In vitro activities of voriconazole, posaconazole, and fluconazole against 4,169 clinical isolates of Candida spp . and Cryptococcus neoformans collected during 2001 and 2002 in the ARTEMIS global antifungal surveillance program; Pfaller MA et al.; We examined the in vitro activities of voriconazole, posaconazole, and fluconazole against 3,932 isolates of Candida spp . and 237 isolates of Cryptococcus neoformans obtained from over 100 medical centers worldwide during 2001 and 2002 . The MICs of the antifungal drugs were determined by broth microdilution tests performed according to the National Committee for Clinical Laboratory Standards (NCCLS) methods using RPMI 1640 as the test medium . Voriconazole and posaconazole were very active against Candida spp . (97-98% susceptible at MICs < or =1 microg/ml) and C . neoformans (98-100% susceptible at MICs < or =1 microg/mL) . C . albicans (MIC90, 0.015-0.03 microg/ml) was the most susceptible species of Candida to both agents and C . glabrata (MIC90, 1-2 microg/mL) was the least susceptible . Both voriconazole and posaconazole were more active than fluconazole against all Candida spp . and C . neoformans . These results provide further evidence for the increased spectrum and potency of the new triazoles against a large and geographically diverse collection of opportunistic fungal pathogens. Indian J Pathol Microbiol, 2003 Apr, 46(2), 207 - 11 Pneumocystis carinii pneumonia and its association with other opportunistic infections in AIDS--an autopsy report of five cases; Deshmukh SD et al.; Pneumocystis carinii pneumonia (PCP) has been reported as one of the opportunistic pathogens in AIDS . The significance of this pathogen in AIDS is well established so that, the diagnosis of PCP in an adult simultaneously establishes the diagnosis of AIDS . This point is well emphasised in the CDC case definition of AIDS . In western literature, the occurrence of PCP in AIDS is widely reported . However, in Indian literature only sporadic case reports have been documented . This study reports 5 cases of PCP encountered amongst 34 AIDS-autopsies studied . PCP alone was present in 2 cases . It is worth noting that it was simultaneously associated with cryptococcosis, tuberculosis and CMV in 3 remaining cases, highlighting the need for extensive investigations even after establishing the diagnosis of PCP in a known full blown AIDS patient. Pediatr Infect Dis J, 2004 Mar, 23(3), 278 - 9 Mother-to-child transmission of cryptococcus neoformans; Sirinavin S et al.; Cryptococcal meningitis was diagnosed in a 92-day-old boy who was not HIV-1-infected and who survived after treatment, although with hydrocephalus . The mother was HIV-1-infected, delivered prematurely, had peripartum cryptococcal meningitis and died 14 days postpartum . There was no other possible source for cryptococcal infection in this infant . This is believed to be a case of mother-to-child transmission of cryptococcosis. J Biol Chem, 2004 May 14, 279(20), 21144 - 53 Epub 2004 Mar 10. The sphingolipid pathway regulates Pkc1 through the formation of diacylglycerol in Cryptococcus neoformans; Heung LJ et al.; The sphingolipid biosynthetic pathway generates bioactive molecules crucial to the regulation of mammalian and fungal physiological and pathobiological processes . In previous studies (Luberto, C., Toffaletti, D . L., Wills, E . A., Tucker, S . C., Casadevall, A., Perfect, J . R., Hannun, Y . A., and Del Poeta, M . (2001) Genes Dev . 15, 201-212), we demonstrated that an enzyme of the fungal sphingolipid pathway, Ipc1 (inositol-phosphorylceramide synthase-1), regulates melanin, a pigment required for the pathogenic fungus Cryptococcus neoformans to cause disease . In this study, we investigated the mechanism by which Ipc1 regulates melanin production . Because Ipc1 also catalyzes the production of diacylglycerol (DAG), a physiological activator of the classical and novel isoforms of mammalian protein kinase C (PKC), and because it has been suggested that PKC is required for melanogenesis in mammalian cells, we investigated whether Ipc1 regulates melanin in C . neoformans through the production of DAG and the subsequent activation of Pkc1, the fungal homolog of mammalian PKC . The results show that modulation of Ipc1 regulates the levels of DAG in C . neoformans cells . Next, we demonstrated that C . neoformans Pkc1 is a DAG-activated serine/threonine kinase and that the C1 domain of Pkc1 is necessary for this activation . Finally, through both pharmacological and genetic approaches, we found that inhibition of Pkc1 abolishes melanin formation in C . neoformans . This study identifies a novel signaling pathway in which C . neoformans Ipc1 plays a key role in the activation of Pkc1 through the formation of DAG . Importantly, this pathway is essential for melanin production with implications for the pathogenicity of C . neoformans. Carbohydr Res, 2004 Feb 25, 339(3), 503 - 9 Structural studies of the capsular polysaccharide of a non-neoformans Cryptococcus species identified as C . laurentii, which was reclassified as Cryptococcus flavescens, from a patient with AIDS; Ikeda R et al.; Cryptococcus flavescens, a strain originally identified as C . laurentii, was isolated from the cerebrospinal fluid of an AIDS patient, and the soluble capsular polysaccharide of the yeast was investigated . Glucuronoxylomannan (GXM) was obtained from C . flavescens under conditions similar to those used to obtain C . neoformans polysaccharide . However, the GXM differed from C . neoformans polysaccharide in the decreased O-acetyl group content . The structure of GXM was determined by methylation analysis, partial acid hydrolysis, NMR analyses, and controlled Smith degradation . These analyses indicated that GXM has the following structure: an alpha-(1-->3)-D-mannan backbone with side chains of beta-D-glucuronic acid residues bound to the C-2 position of the mannose residue . The C-6 position of the mannose is substituted with D-man-beta-(1-->4)-D-xyl-beta-(1--> disaccharide . Furthermore, the existence of side chains containing more than two xylose residues was suggested . This mannosylxylose side chain is a novel structure in polysaccharides of C . neoformans and other Cryptococcus species. Mycopathologia, 2004 Jan, 157(1), 29 - 36 Performance of selective and differential media in the primary isolation of yeasts from different biological samples; Silva JO et al.; In view of the increase in yeast infections, especially polymicrobial ones, differential culture media have acquired increasing importance . The present study evaluated the Sabouraud chloramphenicol, Biggy agar, Pagano Levin agar and CHROMagar Candida media in terms of isolation, number of yeast colony forming units per plate, and inhibition of bacteria and filamentous fungi . To this end, we used 223 biological samples, including feces, and oral, vaginal and anal mucosae from 86 patients presenting or not symptoms of fungal infections . The four media did not differ significantly in terms of detection of yeast-positive cultures . The number of colony forming units per plate ranged from zero to 2.380, with a predominance of counts of 1 to 9 colonies per plate . No significant differences were observed among the four culture media in terms of number of colonies counted, for each kind of biological material . Fifteen species belonging to the genera Candida, Saccharomyces, Cryptococcus, Trichosporon and Rhodotorula were isolated, with C . albicans being the predominant species, followed by C . parapsilosis and R . rubra . CHROMagar Candida and Biggy agar were complementary in the isolation of the different species and favored a greater recovery of polymicrobial cultures . Pagano Levin agar isolated the smallest variety of species . Sabouraud chloramphenicol agar was the least effective in terms of bacterial inhibition and favored a greater development of filamentous fungi . The results suggest that more than one culture medium should be used for an adequate primary isolation. Intern Med, 2004 Feb, 43(2), 117 - 9 Pulmonary cryptococcosis with a solitary focal ground-glass opacity on high-resolution computed tomography; Sato M et al.; We report a 74-year-old woman with cervical cancer who developed pulmonary cryptococcosis which presented as a solitary focal ground-glass opacity (GGO) on high-resolution computed tomography (HRCT) . Serial HRCT showed the progression from the GGO to a discrete solid nodule . We hypothesize that the initial GGO may correspond pathologically to partial filling of air spaces with cryptococcal organisms and inflammatory cells . To our knowledge, this is the first report of pulmonary cryptococcosis with a solitary focal GGO on HRCT in the literature. J Immunol, 2004 Mar 15, 172(6), 3670 - 7 Unexpected diversity in the fine specificity of monoclonal antibodies that use the same V region gene to glucuronoxylomannan of Cryptococcus neoformans; McFadden DC et al.; Most mAbs to the capsular polysaccharide glucuronoxylomannan (GXM) of Cryptococcus neoformans are generated from the same VH and VL gene families . Prior Ab studies have assessed protective efficacy, Id structure and binding to capsular polysaccharides, and peptide mimetics . These data have been interpreted as indicating that most mAbs to GXM have the same specificity . A new approach to Ab specificity analysis was investigated that uses genetic manipulation to generate C . neoformans variants with structurally different capsules . C . neoformans mutants expressing GXM with defective O-acetylation were isolated and complemented by the C . neoformans gene CAS1, which is necessary for the O-acetylation of GXM . The mAbs exhibited differences in their binding to the GXM from these mutant strains, indicating previously unsuspected differences in specificity . Analysis of three closely related IgMs revealed that one (mAb 12A1) bound to an epitope that did not require O-acetylation, another (mAb 21D2) was inhibited by O-acetylation, and the third (mAb 13F1) recognized an O-acetylation-dependent conformational epitope . Furthermore, an IgG Ab (mAb 18B7) in clinical development retained binding to de-O-acetylated polysaccharide; however, greater binding was observed to O-acetylated GXM . Our findings suggest that microbial genetic techniques can provide a new approach for epitope mapping of polysaccharide-binding Abs and suggest that this method may applicable for studying the antigenic complexity of polysaccharide Ags in other capsulated microorganisms. J Clin Microbiol, 2004 Mar, 42(3), 1356 - 9 Cryptococcus neoformans shows a remarkable genotypic diversity in Brazil; Barreto de Oliveira MT et al.; The genotypic diversity of Brazilian Cryptococcus neoformans strains was analyzed . The majority of the samples were alphaA (65%), followed by alphaB (17.5%), alphaD (9%), alphaAaD hybrids (5%), and alphaC (3.5%) . A considerable genotypic diversity occurred within C . neoformans var . grubii, and a new amplified fragment length polymorphism genotype, 1B, was recognized. Ann Otol Rhinol Laryngol, 2004 Feb, 113(2), 121 - 3 Cryptococcal laryngitis: case report and review of the literature; Nadrous HF et al.; Infections caused by Cryptococcus neoformans may range from an asymptomatic illness to lethal systemic disease, especially in immunocompromised hosts . Although cryptococcal infection most commonly involves the lungs or the central nervous system, it can disseminate to virtually any organ . Laryngeal cryptococcal infections are extremely rare--only 5 cases have been reported in the literature . Herein, we describe cryptococcal laryngitis occurring in a 55-year-old man with asthma and allergic fungal sinusitis . He presented with hoarseness and cough . He was treated with itraconazole followed by fluconazole therapy with complete recovery from his laryngeal infection . The patient was disease-free when last seen 10 months after the diagnosis . We present a summary of all reported cases of cryptococcal laryngitis. Photochem Photobiol Sci, 2004 Mar, 3(3), 281 - 6 Epub 2003 Dec 02. Constitutive and UV-inducible synthesis of photoprotective compounds (carotenoids and mycosporines) by freshwater yeasts; Libkind D et al.; Twelve yeasts isolated from lakes of Northwestern Patagonia, Argentina, belonging to eight genera (Sporobolomyces, Sporidiobolus, Rhodotorula, Rhodosporidium, Cystofilobasidium, Cryptococcus, Torulaspora, and Candida) were analysed for their ability to produce photoprotective compounds . For this purpose, three laboratory experiments were performed to study the effect of photosynthetically active radiation (PAR) and PAR in combination with UV radiation (PAR + UVR) on the production of carotenoids and mycosporines . The synthesis of carotenoid compounds was clearly stimulated in six out of nine red yeast strains tested upon exposure to PAR or PAR + UVR; however, the latter conditions produced a stronger response than PAR alone . The increase in carotenoids in the red strains under PAR + UVR irradiation showed a negative exponential relationship with their basal carotenoid content, suggesting that cells with higher constitutive levels of carotenoids are less responsive to induction by PAR + UVR . Three red yeasts, Rhodotorula minuta, Rh . pinicola, and Rhodotorula sp., and the colourless Cryptococcus laurentii produced a UV-absorbing compound when exposed to PAR or PAR + UVR . This compound showed an absorption maximum at 309-310 nm and was identified as mycosporine-glutaminol-glucoside (myc-glu-glu) . In these strains, exposure to PAR or PAR + UVR resulted in elevated concentrations of both carotenoids and myc-glu-glu . This is the first report on the production of mycosporines by yeasts . All strains that developed under PAR + UVR were able to synthesise carotenoids either constitutively or in response to PAR exposure, and a few of them also produced myc-glu-glu when exposed to PAR . Collectively, our results suggest that the presence of carotenoids, either alone or in combination with mycosporines, are required for sustaining growth under exposure to PAR + UVR in the freshwater yeast strains studied. Pharmacotherapy, 2004 Feb, 24(2 Pt 2), 4S - 28S; quiz 29S-32S Changing strategies for the management of invasive fungal infections; Rapp RP; The frequency of invasive fungal infections (IFIs) has increased with the increase in number of high-risk patients . United States trends in mortality due to invasive mycoses showed a striking increase in the past 2 decades . Human immunodeficiency virus-associated opportunistic mycoses accounted for part of the increase, as did mycoses in other immunocompromised populations . Those at-risk populations include recipients of solid organ transplants or hematopoietic stem cell transplants, those with hematologic malignancies, and others receiving immunosuppressive treatment . Infections due to Candida sp are the most common fungal infections . The mortality rate due to invasive aspergillosis has risen steadily, with a 357% increase from 1980-1997 . Depending on whether an IFI is possible, probable, or proved, three treatment strategies are available: prophylactic, empiric, and specific . Options for preventing and treating IFIs have increased, with four classes of antifungal agents available: polyenes, azoles, nucleoside analogs, and echinocandins . The effectiveness of the polyene amphotericin B deoxycholate (the standard for > 40 yrs) is limited by toxic effects, notably renal and infusion-related toxicity . Three lipid formulations are approved for the treatment of IFIs in patients refractory to or intolerant of amphotericin B: amphotericin lipid complex, amphotericin B colloidal dispersion, and liposomal amphotericin B . The nucleoside analog class contains only flucytosine--an antimetabolite targeting the nucleus of the fungal cell and generally not given as monotherapy because of frequent development of resistance . The azoles debuted with ketoconazole, followed by fluconazole, itraconazole, and voriconazole . Fluconazole is largely active against Candida sp and Cryptococcus neoformans; itraconazole's activity is largely against moulds, such as Aspergillus, and dimorphic fungi, such as Histoplasma and Blastomyces; and voriconazole is active against both yeasts and moulds . The echinocandin class has one drug approved for clinical use--caspofungin, which targets the fungal cell wall . Deciding which antifungal agent to use involves weighing such clinical factors as mycoses susceptibility and drug toxicity, as well as pharmacoeconomic considerations . Besides the price of the drug, the cost of antifungal therapy includes costs of mortality associated with failed treatment, prolonged hospitalization and treatment related to complications, and additional antifungal treatment to compensate for primary treatment failure. Pathol Res Pract, 2003, 199(12), 811 - 4 Autopsy findings in AIDS patients from a reference hospital in Brazil: analysis of 92 cases; Cury PM et al.; The aim of this work was to evaluate the opportunistic diseases and the cause of death of AIDS patients who were submitted to autopsy . We included all AIDS patients submitted to autopsy at a reference hospital of a medical school in Sao Paulo, Brazil, during the period of 1993 to 2000 . Out of 1,478 autopsy cases in this period, 92 patients (6.22%) had the previously confirmed diagnosis of AIDS . Sixty-nine patients (75%) were men ranging in age from 19 to 68 years (mean 34.8) . Eighty-five patients (92.4%) died due to infectious diseases, while only two died of neoplasia . Forty-four (48%) patients died from pulmonary infection, 14 (15%) from sepsis, 14 (15%) from disseminated mycobacteriosis, and six (7%) from Central Nervous System (CNS) infection . The opportunistic diseases found were mycobacteriosis (n = 25), Pneumocystis carinii infection (n = 16), Cytomegalovirus infection (n = 17), toxoplasmosis (8 CNS cases), candida sp infection (n = 12), histoplasmosis (n = 5), cryptococcus (n = 4), and one case of blastomycosis in the lung . Most of our AIDS patients are dying of infectious and opportunistic diseases that are not always diagnosed during their lifetime. FEMS Microbiol Lett, 2004 Feb 16, 231(2), 165 - 9 Biochemical characterization of alpha-amylase from the yeast Cryptococcus flavus; Wanderley KJ et al.; During our screening of amylolytic microorganisms from Brazilian fruits, we isolated a yeast strain classified as Cryptococcus flavus . When grown on starch-containing medium this strain exhibited the highest amylase production after 24 h of cultivation . The extracellular amylase from C . flavus was purified from the culture broth by a single step using chromatography on a Sephacryl S-100 column . The enzyme was purified 16.14-fold with a yield of 50.21% of the total activity . The purified enzyme was a glycoprotein with an apparent molecular mass of 75 and 84.5 kDa as estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and gel filtration, respectively . The enzyme lost approximately 50% of the molecular mass after treatment with glycosidases . The major end products of starch, amylose, amylopectin, pullulan and glycogen were maltose and maltotriose . The K(m) value for the pure enzyme was 0.056 mg ml(-1) with soluble starch as the substrate . Enzyme activity was optimal at pH 5.5 and 50 degrees C . The enzyme retained 90% of the activity after incubation at 50 degrees C for 60 min and was inhibited by Cu(2+), Fe(2+) and Hg(2+). Clin Exp Dermatol, 2004 Mar, 29(2), 159 - 60 Primary cutaneous cryptococcosis in an elderly man; Werchniak AE et al.; Cryptococcus neoformans is an opportunistic yeast with a worldwide distribution . Although well recognized as a cause of cutaneous disease in immunocompromised patients, this rarely occurs in healthy hosts . In this article, we present the case of a farmer who developed cutaneous cryptococcosis at the site of an injury he sustained while cleaning out his barn . Despite an extensive work-up, no evidence of disseminated disease or underlying immunosuppression was found . The patient recovered completely several weeks after beginning treatment with oral fluconazole. J Leukoc Biol, 2004 Jun, 75(6), 1147 - 55 Epub 2004 Feb 24. Fcgamma receptor signaling in primary human microglia: differential roles of PI-3K and Ras/ERK MAPK pathways in phagocytosis and chemokine induction; Song X et al.; Cryptococcus neoformans monoclonal antibody immune complex (IC) induces beta-chemokines and phagocytosis in primary human microglia via activation of Fc receptor for immunoglobulin G (FcgammaR) . In this report, we investigated microglial FcgammaR signal-transduction pathways by using adenoviral-mediated gene transfer and specific inhibitors of cell-signaling pathways . We found that Src inhibitor PP2 and Syk inhibitor piceatannol inhibited phagocytosis, macrophage-inflammatory protein-1alpha (MIP-1alpha) release, as well as phosphorylation of extracellular-regulated kinase (ERK) and Akt, consistent with Src/Syk involvement early in FcgammaR signaling . Constitutively active mitogen-activated protein kinase kinase (MEK) induced MIP-1alpha, and Ras dominant-negative (DN) inhibited IC-induced ERK phosphorylation and MIP-1alpha production . These results suggest that the Ras/MEK/ERK pathway is necessary and sufficient in IC-induced MIP-1alpha expression . Neither Ras DN nor the MEK inhibitor U0126 inhibited phagocytosis . In contrast, phosphatidylinositol-3 kinase (PI-3K) inhibitors Wortmannin and LY294002 inhibited phagocytosis without affecting ERK phosphorylation or MIP-1alpha production . Conversely, Ras DN or U0126 did not affect Akt phosphorylation . Together, these results demonstrate distinct roles played by the PI-3K and Ras/MEK/ERK pathways in phagocytosis and MIP-1alpha induction, respectively . Our results demonstrating activation of functionally distinct pathways following microglial FcgammaR engagement may have implications for human central nervous system diseases. Antimicrob Agents Chemother, 2004 Mar, 48(3), 1004 - 6 Evaluation of acute hematologic and long-term pulmonary toxicities of radioimmunotherapy of Cryptococcus neoformans infection in murine models; Dadachova E et al.; We evaluated acute hematological and long-term pulmonary toxicity of radioimmunotherapy in murine models of Cryptococcus neoformans infection . Activities up to 250 microCi were well tolerated by healthy A/JCr mice for (213)Bi-18B7 and (188)Re-18B7 monoclonal antibodies . In infected mice, doses up to 150 microCi produced only transient toxicity . The lungs of treated mice had no evidence of radiation fibrosis. Antimicrob Agents Chemother, 2004 Mar, 48(3), 985 - 91 Amphotericin B and fluconazole, a potent combination therapy for cryptococcal meningitis; Larsen RA et al.; We evaluated the antifungal activities of amphotericin B, fluconazole, and flucytosine, alone and in combination, in a murine model of cryptococcal meningitis . The objectives were to determine the greatest antifungal effects achievable with these drugs alone or in combination . Meningitis was established in male BALB/c mice weighing 23 to 25 g by intracerebral injection of Cryptococcus neoformans . Treatment was started on day 2 . Amphotericin B was tested at 0.3 to 1.3 mg/kg of body weight/day by slow intravenous injection . Fluconazole at 10 to 40 mg/kg/day and flucytosine at 20 to 105 mg/kg/day were administered in the sole source of drinking water . The mice were killed at 16 days, and the numbers of fungal colonies in the brain were quantified . The association between the response and the dose combination was evaluated by local nonparametric response surface methods; 99% confidence intervals were used to evaluate the antifungal effects . Ninety-five percent of the mice treated with amphotericin B at 0.5 mg/kg survived to the end of the experiment, regardless of the fluconazole or flucytosine dose used . The greatest activity was seen with amphotericin B plus fluconazole with or without flucytosine . However, the addition of flucytosine did not increase the antifungal activity . Given the widespread availability of amphotericin B and fluconazole and the relative safety profile of fluconazole compared to that of flucytosine, the full potential of this two-drug combination deserves further evaluation. Mol Microbiol, 2004 Mar, 51(5), 1447 - 58 Thiol peroxidase is critical for virulence and resistance to nitric oxide and peroxide in the fungal pathogen, Cryptococcus neoformans; Missall TA et al.; Cryptococcus neoformans is a fungal pathogen most commonly causing meningitis in immunocompromised patients . Current therapies are inadequate, and novel antifungal targets are needed . We have identified by proteomics two thiol peroxidases that are differentially expressed at 37 degrees C, the temperature of the mammalian host . Consistent with their antioxidant role, we show that the genes encoding these thiol-specific antioxidants, TSA1 and TSA3, are transcriptionally induced when C . neoformans is exposed to hydrogen peroxide . Genome sequence analysis of C . neoformans revealed a third thiol peroxidase, TSA4 . We constructed single, double and triple mutants of the thiol peroxidase genes through homologous recombination and analysed their function by comparing the growth of these mutants with that of the wild-type strain . The tsa1 Delta mutant shows sensitivity to hydrogen peroxide and t-butylhydroperoxide, as well as significant growth retardation at 25 degrees C and 38.5 degrees C . The tsa1 Delta mutant is also sensitive to NO, demonstrating a link between oxidative and nitrosative stress pathways . In two mouse models of cryptococcosis, the tsa1 Delta mutant is significantly less virulent. Med Mycol, 2004 Feb, 42(1), 15 - 25 Resistance to Cryptococcus neoformans infection in the absence of CD4+ T cells; Aguirre K et al.; Previous studies of Cryptococcus neoformans infection have revealed a role for CD4+ T cells and CD8+ T cells in anticryptococcal resistance in the lungs, but such a role has been revealed only for CD4+ T cells in the brains of experimentally infected mice . In this study, we found that mice genetically engineered to lack CD4+ T cells could be successfully vaccinated to express resistance to a rechallenge with Cryptococcus neoformans, provided the challenge dose was kept to lower than 1000 organisms per mouse . The challenge infection was uniformly lethal for unvaccinated control mice . Depletion of CD8+ T cells weakened this resistance to re-challenge: both naive and vaccinated mice that were treated with antibody raised against CD8+ T cells died significantly earlier than did mice that received an irrelevant control antibody . In vitro, purified CD8+ T cells taken from draining lymph nodes of antigen-experienced mice were less efficient than were identically prepared CD4+ T cells at stimulating the cells of a transformed microglial cell line to inhibit C . neoformans proliferation, possibly mirroring the inferiority of CD8+ T-cell-mediated protection observed in vivo . RNase protection assays showed similar IFN-gamma mRNA levels in both lymphocyte subsets . Class II major histocompatibility antigen expression was up-regulated strikingly on microglia cultured with IFN-gamma, but class I expression was less dramatically affected . Therefore microglial cell interaction may be more greatly enhanced with CD4+ cells than with CD8+ cells. Infect Immun, 2004 Mar, 72(3), 1746 - 54 Protective efficacy of antigenic fractions in mouse models of cryptococcosis; Mansour MK et al.; Infections due to the encapsulated fungus Cryptococcus neoformans are a significant cause of morbidity and mortality in patients with impaired T-cell function, particularly those with AIDS . Presumably then, T-cell responses to cryptococcal antigens are critical for protection against this ubiquitous fungus . To test the protective efficacy of these antigens as vaccine candidates, secreted cryptococcal antigens were separated by concanavalin A affinity chromatography into adherent (mannoprotein {MP}) and nonadherent (flowthrough {FT}) fractions, and the fractions were tested in murine models of disseminated cryptococcosis . Compared with adjuvant alone, C57BL/6 mice that received two inoculations of MP and FT exhibited prolonged survival and reduced brain and kidney fungal loads following intravenous challenge with C . neoformans strain B3501 . MP-immunized animals had increased brain levels of tumor necrosis factor alpha, gamma interferon, and interleukin-2 . Histopathologic examination revealed that compared with organs from mice that received only adjuvant, MP-immunized mice were able to recruit a stronger cellular infiltrate in brain, kidney, and liver in response to cryptococcal infection . Conjugated O-linked glycans were necessary for optimal MP-mediated protection, because chemical O deglycosylation reduced the protective efficacy of MP immunization . FT and MP immunization protected B-cell-deficient, but not T-cell-deficient mice, suggesting that protection was T-cell mediated . CBA/J mice also benefited from immunization with FT and MP, although the benefits were more modest than those seen with C57BL/6 mice . Thus, both MP and FT fractions of C . neoformans contain components that protect mice from disseminated cryptococcosis, and this protection appears to be T-cell mediated. Infect Immun, 2004 Mar, 72(3), 1693 - 9 CNLAC1 is required for extrapulmonary dissemination of Cryptococcus neoformans but not pulmonary persistence; Noverr MC et al.; The pathogenic yeast Cryptococcus neoformans produces a laccase enzyme (CNLAC1), which catalyzes the synthesis of melanin in the presence of phenolic compounds . A number of genes have been implicated in the regulation of laccase and melanization, including IPC1, GPA1, MET3, and STE12 . Albino mutants derived from random mutagenesis techniques may contain mutations in genes that regulate multiple virulence factors, including CNLAC1 . The goal of our study is to investigate the role of CNLAC1 in virulence and evasion of pulmonary host defenses after infection via the respiratory tract . Using a set of congenic laccase-positive (2E-TUC-4) and laccase-deficient (2E-TU-4) strains, we found that both strains are avirulent at a lower dose (10(4) CFU/mouse) in mice . After the infectious dose was increased to 10(6) CFU/mouse, 70% mortality was observed in mice infected with 2E-TUC-4 compared to no mortality in mice infected with 2E-TU-4 at day 30 postinfection . This observation confirms the requirement for CNLAC1 in virulence . Interestingly, we observed no differences between the two strains in pulmonary growth or in elicitation of cellular immune responses in the lung . The only measurable defect of 2E-TU-4 was in dissemination to extrapulmonary sites . To examine the role of CNLAC1 in dissemination, mice were infected intravenously . By week 3 postinfection, equal numbers of strains 2E-TUC-4 and 2E-TU-4 were recovered from the brain and spleen . This observation indicates that CNLAC1 facilitates escape from the lung, but not growth in the lungs or brain, and suggests a novel role for CNLAC1 in virulence during an infection aquired via the respiratory tract. Salud Publica Mex, 2003 Nov-Dec, 45(6), 455 - 60 {Frequency of mycosis in immunosuppressed patients at a regional hospital of Mexico City}; Hernandez-Hernandez F et al.; OBJECTIVE: To isolate and identify the fungi in specimens collected from immunocompromised patients seen in Mexico City hospital, and to assess their association with mycosis . MATERIAL AND METHODS: A total of 268 specimens from 108 patients were processed for mycological study, including direct examination, smears, cultures, and microcultures in specific media, in addition to biochemical tests . Most of the patients had been diagnosed with pulmonary tuberculosis and Acquired Immunodeficiency Syndrome (AIDS) . RESULTS: One hundred and eighty-three yeasts and 66 mycelial fungi were isolated . Forty-five mycoses were diagnosed; the most frequent mycosis was pulmonary candidosis (32 cases) . Candida albicans and C parapsilosis were the predominant species associated with mycotic infection . Five Cryptococcus neoformans var . neoformans, one C albidus, three Histoplasma capsulatum and one Geotrichum candidum isolates were associated with mycosis . CONCLUSIONS: Of the studied patients, 41.6% showed a mycotic infection, principally associated with AIDS and pulmonary tuberculosis . The English version of this paper is available at:http://www.insp.mx/salud/index.html. Hong Kong Med J, 2004 Feb, 10(1), 38 - 43 Screening for vestibular schwannoma by magnetic resonance imaging: analysis of 1821 patients; Kwan TL et al.; OBJECTIVES: To study the spectrum of diseases that can be detected by magnetic resonance imaging in patients suspected to have vestibular schwannoma (acoustic neuroma) presenting with sensorineural or mixed hearing loss, and to assess the extent of the problem of hearing loss in a screened population . DESIGN: Retrospective study . SETTING: Diagnostic radiology and imaging department of a regional hospital, Hong Kong . PATIENTS: A total of 1821 consecutive patients from September 1999 to February 2001 with sensorineural or mixed hearing loss were referred by otolaryngologists for magnetic resonance imaging of the internal auditory canal . MAIN OUTCOME MEASURES: Vestibular schwannoma; other cerebellopontine angle masses and other diseases that could account for the patients' hearing loss . RESULTS: In all, 132 (7%) patients had positive findings that could explain their hearing loss . Fifty-four (41%) of the 132 patients had vestibular schwannoma; 39 (30%) had inflammation of the middle ear and mastoids; 17 (13%) had ischaemic foci in the brainstem; 10 (8%) had other cerebellopontine angle masses or tumours; four (3%) had inner ear dysplasia; seven (5%) had vascular loop compression; and one (1%) had chronic cryptococcal meningitis . The overall incidence of vestibular schwannoma detected in this screened population was about 3% . CONCLUSIONS: This study indicates that magnetic resonance imaging is an effective tool to screen for vestibular schwannoma in patients with sensorineural or mixed hearing loss . It can also be used to assess a considerable number of different pathological conditions in patients with audiovestibular disorders. J Nucl Med, 2004 Feb, 45(2), 313 - 20 Susceptibility of the human pathogenic fungi Cryptococcus neoformans and Histoplasma capsulatum to gamma-radiation versus radioimmunotherapy with alpha- and beta-emitting radioisotopes; Dadachova E et al.; Fungal diseases are difficult to treat in immunosuppressed patients and, consequently, new approaches to therapy are urgently needed . One novel strategy is to use radioimmunotherapy (RIT) with fungal-binding monoclonal antibodies (mAbs) labeled with radionuclides . However, many fungi manifest extreme resistance to gamma-radiation, such that the doses of several thousand gray are required for 90% cell killing, whereas for mammalian cells the lethal dose is only a few gray . We compared the susceptibility of human pathogenic fungi Cryptococcus neoformans (CN) and Histoplasma capsulatum (HC) to external gamma-radiation and to the organism-specific mAbs 18B7 and 9C7, respectively, conjugated to (213)Bi and (188)Re radionuclides . METHODS: CN and HC cells were irradiated with up to 8,000 Gy ((137)Cs source, 30 Gy/min) . RIT of CN with (213)Bi- and (188)Re-labeled specific mAb and of HC with (188)Re-labeled specific mAb used 0-1.2 MBq per 10(5) microbial cells . After irradiation or RIT, the cells were plated for colony-forming units (CFUs) . Cellular dosimetry calculations were performed, and the pathway of cell death after irradiation was evaluated by flow cytometry . RESULTS: Both CN and HC proved to be extremely resistant to gamma-radiation such that significant killing was observed only for doses of >4,000 Gy . In contrast, these cells were much more susceptible to killing by radiation delivered with a specific mAb, such that a 2-logarithm reduction in colony numbers was achieved by incubating them with (213)Bi- and (188)Re-labeled mAb 18B7 or with (188)Re-9C7 mAb . Dosimetry calculations showed that RIT was approximately 1,000-fold more efficient in killing CN and approximately 100-fold more efficient in killing HC than gamma-radiation . Both gamma-radiation and RIT caused cell death via an apoptotic-like pathway with a higher percentage of apoptosis observed in RIT-treated cells . CONCLUSION: Conjugating a radioactive isotope to a fungal-specific antibody converted an immunoglobulin with no antifungal activity into a microbicidal molecule . RIT of fungal cells using specific antibodies labeled with alpha- and beta-emitting radioisotopes was significantly more efficient in killing CN and HC than gamma-radiation when based on the mean absorbed dose to the cell . These results strongly support the concept of using RIT as an antimicrobial modality. Eukaryot Cell, 2004 Feb, 3(1), 14 - 26 Cyclic AMP-dependent protein kinase catalytic subunits have divergent roles in virulence factor production in two varieties of the fungal pathogen Cryptococcus neoformans; Hicks JK et al.; Our earlier findings established that cyclic AMP-dependent protein kinase functions in a signaling cascade that regulates mating and virulence of Cryptococcus neoformans var . grubii (serotype A) . Mutants lacking the serotype A protein kinase A (PKA) catalytic subunit Pka1 are unable to mate, fail to produce melanin or capsule, and are avirulent in animal models, whereas mutants lacking the PKA regulatory subunit Pkr1 overproduce capsule and are hypervirulent . Because other mutations have been observed to confer different phenotypes in two diverged varieties of C . neoformans (grubii variety {serotype A} and neoformans variety {serotype D}), we analyzed the functions of the PKA genes in the serotype D neoformans variety . Surprisingly, the Pka1 catalytic subunit was not required for mating, haploid fruiting, or melanin or capsule production of serotype D strains . Here we identify a second PKA catalytic subunit gene, PKA2, that is present in both serotype A and D strains of C . neoformans . The divergent Pka2 catalytic subunit was found to regulate mating, haploid fruiting, and virulence factor production in serotype D strains . In contrast, Pka2 has no role in mating, melanin production, or capsule formation in serotype A strains . Our studies illustrate how different components of signaling pathways can be co-opted and functionally specialized during the evolution of related but distinct varieties or subspecies of a human fungal pathogen. Int J STD AIDS, 2004 Jan, 15(1), 48 - 50 Aetiology of meningitis in HIV-infected patients in a referral hospital in Phnom Penh, Cambodia; Chhin S et al.; The HIV epidemic in Cambodia is one of the most extensive in Asia . Meningitis accounts for a substantial proportion of HIV-related morbidity and mortality in Cambodia . A retrospective chart review was performed to identify the clinical and spinal fluid characteristics of patients undergoing spinal tap at an AIDS referral hospital in Phnom Penh, Cambodia during a 16-month period . Of 932 charts reviewed, 89 met criteria for analysis . Overall mortality was 49.4% . Cryptococcus was the most commonly identified pathogen (83%), followed by mycobacteria (8%) . No pathogen was identified in 9% of charts reviewed . In hospital mortality was similar in all groups. Mikrobiologiia, 2003 Nov-Dec, 72(6), 822 - 7 {A new basidiomycetous yeast species, Cryptococcus mycelialis, related to Holtermannia Saccardo et Traverso}; Golubev VI et al.; A new species of the genus Cryptococcus, Cr . mycelialis (the type strain VKM Y-2863), is described based on the taxonomic study of four strains isolated from the soil and plant samples collected on the South Georgia and East Falkland islands . This species differs from the known Cryptococcus species in its ability to form a true monokaryotic mycelium with pseudoclamp connections and haustoria . The species can be distinguished from the phylogenetically related and phenotypically similar species Holtermannia corniformis and Cr . nyarrowii by some assimilatory reactions, maximum growth temperature, and sensitivity to mycocins. J Clin Microbiol, 2004 Feb, 42(2), 722 - 6 Purification and characterization of a 43-kilodalton extracellular serine proteinase from Cryptococcus neoformans; Yoo Ji J et al.; An extracellular proteinase was purified from culture filtrates of Cryptococcus neoformans NHPY24 by DEAE ion-exchange chromatography and gelatin affinity column chromatography with azoalbumin as the substrate . The molecular mass of the purified enzyme was 43 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, its pH optimum was 7.0 to 8.0, and maximal activity was obtained at pH 7.5 and 37 degrees C . By isoelectric focusing, the purified enzyme had a pI of 4.77 . Enzyme activity was inhibited by serine proteinase inhibitors such as phenylmethylsulfonyl fluoride and diisopropylfluorophosphate . The purified enzyme was thus a serine proteinase . It hydrolyzed natural substrates including hemoglobin, beta-casein, and gamma globulin. Clin Infect Dis, 2004 Feb 15, 38(4), 565 - 71 Epub 2004 Jan 29. Discontinuation of maintenance therapy for cryptococcal meningitis in patients with AIDS treated with highly active antiretroviral therapy: an international observational study; Mussini C et al.; We conducted a retrospective, multicenter study evaluating the safety of discontinuing maintenance therapy for cryptococcal meningitis after immune reconstitution . Inclusion criteria were a previous definitive diagnosis of cryptococcal meningitis, a CD4 cell count of >100 cells/microL while receiving highly active antiretroviral therapy (HAART), and the subsequent discontinuation of maintenance therapy for cryptococcal meningitis . The primary end point was relapse of cryptococcal disease . As of July 2002, 100 patients were enrolled . When maintenance therapy was discontinued, the median CD4 cell count was 259 cells/microL and the median plasma virus load was <2.30 log10 copies/mL, and serum cryptococcal antigen was undetectable in 56 patients . During a median follow-up period of 28.4 months (range, 6.7-64.5; 262 person-years), 4 events were observed (incidence, 1.53 events per 100 person-years; 95% confidence interval, 0.42-3.92) . Three of these patients had a CD4 cell count of >100 cells/microL and a positive serum cryptococcal antigen test result during the recurrent episode . In conclusion, discontinuation of maintenance therapy for cryptococcal meningitis is safe if the CD4 cell count increases to >100 cells/microL while receiving HAART . Recurrent cryptococcal infection should be suspected in patients whose serum cryptococcal antigen test results revert back to positive after discontinuation of maintenance therapy. Nippon Ishinkin Gakkai Zasshi, 2004, 45(1), 1 - 6 Construction of a complete URA5 deletion strain of a human pathogenic yeast Cryptococcus neoformans; Drivinya A et al.; Cryptococcus neoformans is an opportunistic human pathogen, which infects the central nervous system causing the fatal disease, meningitis . In order to understand the genetic background of this human pathogen, the basic molecular manipulation techniques of deletion, overexpression, and so on have been developed . URA5, a gene encoding orotate phosphoribosyltransferase, has frequently been used to introduce foreign gene fragments by complementing ura5 mutant strains, which are not, however, stable; reversion to uracil prototroph is thus frequently observed on selective condition . The high possibility of reversion makes it inconvenient to use this mutation to identify appropriate transformants and thus, manipulation in molecular genetics . We report here the isolation of a stable ura5 mutant of C . neoformans, designated as TAD1, by eliminating the URA5 gene by homologous recombination using the biolistic DNA delivery system . The availability of the stable ura5 mutant offers the advantage that no spontaneous reversion occurs so that a satisfactory rate of homologous recombination can be achieved . The strain will allow efficient genomic analysis in C . neoformans. Rev Inst Med Trop Sao Paulo, 2003 Nov-Dec, 45(6), 299 - 305 Epub 2004 Jan 26. Cryptococcosis: a review of the Brazilian experience for the disease; Pappalardo MC et al.; Cryptococcosis is a systemic mycosis caused by Cryptococcus neoformans . The disease occurs in patients with cellular immunodeficiency . The incidence of cryptococcosis arises with aids, and mycosis is one of the opportunistic infections that defines AIDS . After the HAART era the occurrence of cryptococcosis decreased all over the world, but it still continues to be a prevalent disease in Brazil . Thus, we consider this paper to be very important as a result of our reviewing of Brazilian literature regarding some relevant aspects of that disease. Jpn J Thorac Cardiovasc Surg, 2004 Jan, 52(1), 21 - 5 Lung adenocarcinoma with coexisting pulmonary cryptococcoma; Kawasaki H et al.; A 73-year-old female was referred to our hospital in June 2000 for the evaluation of an abnormal shadow in the left upper lobe of her lung and two other shadows in the left lower lobe . A computed tomography examination revealed an ill-defined tumor with ground glass opacity in the left upper division segment, suggesting a primary lung adenocarcinoma, and two well-defined nodules in the left lower lobe, suggesting metastatic carcinomas . A transbronchial lung biopsy of the tumor in the upper division segment confirmed the presence of an adenocarcinoma but failed to provide a histological diagnosis for the nodules in the lower lobe . A partial resection of the left lower lobe using video-assisted thoracoscopy revealed granulomatous tissue with no signs of malignancy . A curative resection of the adenocarcinoma in the left upper division segment was therefore performed . Histologically, the resected specimens obtained from the left lower lobe were diagnosed as cryptococcomas . The patient was diagnosed as having a stage IA lung adenocarcinoma and two cryptococcomas . The patient is presently well and has not experienced any recurrences or relapse for 3 years since the resection. Br J Neurosurg, 2003 Dec, 17(6), 556 - 9 Vertebral cryptococcosis simulating tuberculosis; Gupta SK et al.; Infection with the fungus Cryptococcus neoformans is seen predominantly in two forms: (a) pulmonary and (b) cerebromeningeal . Skeletal cryptococcosis is uncommon . There have been only occasional case reports of thoracic vertebral cryptococcosis presenting as cord compression . A young female had cervical lymphadenopathy diagnosed as tuberculosis by fine needle aspiration cytology (FNAC) and was on antitubercular therapy (ATT) for 5 months . She developed rapidly progressive paraparesis and imaging demonstrated a destructive vertebral body lesion involving T2-3 with a paraspinal abscess producing cord compression . A costotransversectomy with excision of the diseased bone and bone grafting was done . Histopathological examination revealed cryptococcosis . The patient was put on antifungal medication, but expired 2 weeks after surgery . Radiological, magnetic resonance imaging and surgical finding of vertebral cryptococcosis can mimic tuberculosis . The definite diagnosis of cryptococcosis depends upon microscopic identification of the organism . A high index of suspicion leading to early surgical confirmation and institution of anti fungal therapy is necessary to reduce the mortality and morbidity. Curr Med Chem, 2004 Jan, 11(2), 253 - 66 Effects of the capsular polysaccharides of Cryptococcus neoformans on phagocyte migration and inflammatory mediators; Ellerbroek PM et al.; An important virulence factor of the pathogenic fungus Cryptococcus neoformans is its polysaccharide capsule . The capsular polysaccharides glucuronoxylomannan (GXM), galactoxylomannan (GalXM) and the mannoproteins (MPs) display various immunomodulatory effects on the host response, such as the inhibition of phagocytosis, suppression of T-cell mediated immunity, and induction of immunogenic tolerance . Moreover, these capsular polysaccharides are able to interfere with the migration of phagocytes despite adequate stimulation of chemokine production and their concerted action accounts for the mild inflammatory response often observed in cryptococcosis . Different mechanisms contribute to this phenomenon . First, cryptococcal polysaccharides impair leukocyte migration towards chemoattractants . A combination of the intrinsic chemoattracting properties of circulating polysaccharides and the ability to induce cross-desensitization of chemokine receptors prevents leukocytes from leaving the bloodstream and migrating towards inflammatory site . Polysaccharide-induced repressive effects on the C5a receptor expression on neutrophils may also add to this impaired chemokinesis . Second, polysaccharides interfere with leukocyte adhesion to and migration through the endothelium . Both GXM and MP-4 induce L-selectin shedding from the surface of leukocytes; hence, interference with leukocyte rolling on the endothelium can be expected . GXM also interferes with the subsequent process of firm leukocyte adhesion to the endothelium in vitro . Thirdly, capsular polysaccharides enhance the production of anti-inflammatory interleukin-10 (IL-10) and induce tumor necrosis factor-alpha (TNFalpha) receptor loss from the surface of neutrophils . The capacity to reduce neutrophil influx makes cryptococcal polysaccharides interesting compounds to study in clinical models of inflammation (i.e.; sepsis, auto-immune disorders) in which leukocyte influx can be potentially damaging to host tissues. Clin Microbiol Infect, 2004 Mar, 10 Suppl 1, 48 - 66 Infections due to emerging and uncommon medically important fungal pathogens; Walsh TJ et al.; The emergence of less common but medically important fungal pathogens contributes to the rate of morbidity and mortality, especially in the increasingly expanding population of immunocompromised patients . These pathogens include septate filamentous fungi (e.g., Fusarium spp., Scedosporium spp., Trichoderma spp.), nonseptate Zygomycetes, the endemic dimorphic pathogen Penicillium marneffei, and non-Cryptococcus, non-Candida pathogenic yeast (e.g., Trichosporon spp.) . The medical community is thus called upon to acquire an understanding of the microbiology, epidemiology and pathogenesis of these previously uncommon pathogens in order to become familiar with the options for prevention and treatment. Zhonghua Er Ke Za Zhi, 2003 Jul, 41(7), 542 - 5 {Causes and diagnostic procedure of diffuse lung disease in 28 children}; Zhao SY et al.; OBJECTIVE: Diffuse lung disease comprises a large, heterogeneous group of pulmonary interstitial and parenchymal disease . It is therefore difficult to some extent to make etiologic diagnosis . Little information on clinical spectrum and diagnostic evaluation of pediatric diffuse lung disease is available in our country . The purpose of this study was to explore the causes of and diagnostic approach to diffuse lung disease in children . METHODS: Twenty-eight children with diffuse lung disease aged 2 months to 14 years were studied retrospectively . Their history, physical examination, radiographic findings, final diagnosis and diagnostic processes were reviewed . RESULTS: Confirmed diagnosis was established in 25 cases and suggestive diagnosis in 3 cases . Confirmed diagnoses included: mycoplasma pneumonia in 1 case, Chlamydia trachomatis pneumonia in 2 cases, Epstein-Barr virus pneumonia in 1, CMV pneumonia in 2, hematogenous disseminated pulmonary tuberculosis in 3, pulmonary cryptococcosis in 1, invasive pulmonary aspergillosis in 2, Staphylococcus aureus sepsis in 1, diffuse bronchiectasis in 2, idiopathic pulmonary hemosiderosis in 1, idiopathic pulmonary fibrosis in 1, extrinsic allergic alveolitis in 1, HIV-related lymphocytic interstitial pneumonitis in 1, Wegner's granulomatosis in 1, Langerhan's cell histiocytosis in 2, and lymphoma in 3 . Suggestive diagnoses included Nocardia pneumonia in 1, Pneumocystis carinii pneumonia in 1, and juvenile rheumatoid arthritis-associated pulmonary fibrosis in 1 . The diagnostic directions of 26 patients were conducted by radiographic features . In 17 of 26 cases, the diagnostic range was confined by history . The diagnosis of 14 cases was made by noninvasive tests including antibody detection, bacterial culture, those of 8 cases by examination of biopsy material, and those of 2 cases by autopsy . CONCLUSIONS: The causes of pediatric diffuse lung disease included pulmonary infectious disease, idiopathic pulmonary disease and pulmonary lesion associated with systemic diseases . The diagnosis may be made by radiography, history, physical examination, noninvasive tests in most cases, while in some cases invasive procedures were necessary. Stereotact Funct Neurosurg, 2003, 81(1-4), 5 - 9 Stereotactic brainstem biopsy is indicated for the diagnosis of a vast array of brainstem pathology; Samadani U et al.; BACKGROUND/AIMS: A meta-analysis of thirteen studies performing stereotactic biopsy of brainstem lesions is combined with our recent series of 12 patients with brainstem lesions comparing MRI findings to biopsy histopathology to determine whether these lesions could have been diagnosed radiographically . METHODS: 12 consecutive stereotactic biopsies and literature analysis were performed to analyze clinical, radiographic and histopathologic data . RESULTS: Stereotactic biopsy of lesions in the brainstem was in 96% diagnostic . There was one mortality (3% of cases) and 4% morbidity associated with the procedure . Pathology showed that half of the adult brainstem intrinsic lesions were gliomas, 10% were metastases, and the remainder were hematomas, vascular malformations, lymphomas, demyelination, cysts, radiation necrosis, abscesses, vasculitis, infarcts, leukemia, cryptococcus, or granulomas . CONCLUSIONS: Empiric treatment of adult brainstem lesions is not prudent because there is a wide spectrum of diverse pathology in this location . Stereotactic biopsy is a safe and effective method for determining histopathology . Int J Syst Evol Microbiol, 2004 Jan, 54(Pt 1), 275 - 7 A novel psychrotolerant member of the hymenomycetous yeasts from Antarctica: Cryptococcus watticus sp . nov; Guffogg SP et al.; Soil, snow and organic material, collected in November 1997 from the Vestvold Hills, Davis Base, Antarctica, were screened for yeasts . Two isolates, which were shown to be indistinguishable by rDNA sequencing and protein analysis by SDS-PAGE, are described in this communication as a novel species, Cryptococcus watticus sp . nov . (type culture, CBS 9496T=NRRL Y-27556T) . Sequence analyses of the 26S rDNA D1/D2 region placed C . watticus in the hymenomycetous yeasts in a cluster with Holtermannia corniformis and Cryptococcus nyarrowii . This species has been allocated to the genus Cryptococcus on the basis of physiological and morphological characteristics. Antimicrob Agents Chemother, 2004 Feb, 48(2), 561 - 7 Differential gene expression in auristatin PHE-treated Cryptococcus neoformans; Woyke T et al.; The antifungal pentapeptide auristatin PHE was recently shown to interfere with microtubule dynamics and nuclear and cellular division in the opportunistic pathogen Cryptococcus neoformans . To gain a broader understanding of the cellular response of C . neoformans to auristatin PHE, mRNA differential display (DD) and reverse transcriptase PCR (RT-PCR) were applied . Examination of approximately 60% of the cell transcriptome from cells treated with 1.5 times the MIC (7.89 micro M) of auristatin PHE for 90 min revealed 29 transcript expression differences between control and drug-treated populations . Differential expression of seven of the transcripts was confirmed by RT-PCR, as was drug-dependent modulation of an additional seven transcripts by RT-PCR only . Among genes found to be differentially expressed were those encoding proteins involved in transport, cell cycle regulation, signal transduction, cell stress, DNA repair, nucleotide metabolism, and capsule production . For example, RHO1 and an open reading frame (ORF) encoding a protein with 91% similarity to the Schizophyllum commune 14-3-3 protein, both involved in cell cycle regulation, were down-regulated, as was the gene encoding the multidrug efflux pump Afr1p . An ORF encoding a protein with 57% identity to the heat shock protein HSP104 in Pleurotus sajor-caju was up-regulated . Also, three transcripts of unknown function were responsive to auristatin PHE, which may eventually contribute to the elucidation of the function of their gene products . Further study of these differentially expressed genes and expression of their corresponding proteins are warranted to evaluate how they may be involved in the mechanism of action of auristatin PHE . This information may also contribute to an explanation of the selectivity of auristatin PHE for C . neoformans . This is the first report of drug action using DD in C . neoformans. Antimicrob Agents Chemother, 2004 Feb, 48(2), 384 - 7 In vitro and in vivo efficacies of the new triazole albaconazole against Cryptococcus neoformans; Miller JL et al.; The activity of albaconazole (UR-9825; J . Uriach & Cia . S.A., Barcelona, Spain) was compared to that of fluconazole against 12 isolates of Cryptococcus neoformans in vitro and against 1 isolate in vivo in a rabbit model of cryptococcal meningitis . Albaconazole was 100-fold more potent in vitro than fluconazole on a per-weight basis and was fungicidal at potentially relevant concentrations for two isolates . MICs ranged from </=0.0012 to 1.25 micro g/ml, with the MICs for most isolates being between 0.039 and 0.156 micro g/ml . Isolates were from human immunodeficiency virus (HIV)-infected and non-HIV-infected patients and were of serotypes A, B, and C; and the fluconazole MICs for some of the isolates were elevated . Infected rabbits were treated with either fluconazole or albaconazole at dosages ranging from 5 to 80 mg/kg of body weight/day . The peak concentrations of albaconazole in serum and cerebrospinal fluid (CSF) averaged 4.14 and 0.62 micro g/ml, respectively, in animals receiving 80 mg/kg/day . Comparison of the concentrations in serum and CSF suggested a level of CSF penetration of approximately 15% . Despite limited penetration into the subarachnoid space, at all three doses tested albaconazole was as effective as fluconazole for the treatment of cryptococcal meningitis in rabbits. Bioorg Med Chem Lett, 2004 Feb 9, 14(3), 773 - 7 Bile acid amides derived from chiral amino alcohols: novel antimicrobials and antifungals; Hazra B et al.; Cholic and deoxycholic acid amides 10-17 have been synthesised from (1R,2R)-1-phenyl-2-amino-1,3-propanediol 2, (1S,2S)-1-phenyl-2-amino-1,3-propanediol 4, (1R,2R)-1-para-nitrophenyl-2-amino-1,3-propanediol 3, (1S,2S)-1-para-nitrophenyl-2-amino-1,3-propanediol 5 . Amide 12 derived from N-succinimidyl ester 9 of deoxycholic acid and (1R,2R)-1-phenyl-2-amino-1,3-propanediol 2, found to be active against Cryptococcus neoformans and the amide 17 obtained from N-succinimidyl ester 9 of deoxycholic acid and (1S,2S)-1-para-nitrophenyl-2-amino-1,3-propanediol 5, is found to be potent against various gram-positive bacteria. FEMS Immunol Med Microbiol, 2004 Jan 15, 40(1), 63 - 70 Disruption of CD40/CD40L interaction influences the course of Cryptococcus neoformans infection; Pietrella D et al.; CD40 signaling has been implicated in various pathogenic processes such as chronic inflammatory disease, graft-versus-host disease, autoimmune disease and cancer . We previously demonstrated in an in vitro system that the CD40/CD40L pathway mediates late interleukin (IL) 12 production in response to Cryptococcus neoformans . The purpose of this study was to examine the course of C . neoformans infection in the absence of CD40/CD40L costimulation . We compared infection in mice genetically lacking CD40L (CD40L(-/-)) and in the wild-type counterpart . The animals were injected intratracheally with C . neoformans and monitored for clearance of the organism and the development of cellular immune response . CD40L(-/-) mice exhibited an exacerbation of infection, evaluated as scarce inflammatory response in the lung, that mirrored an increase of fungal burden . This correlated with impairment of nitrite production and antimicrobial activity by macrophages against C . neoformans and unrelated microorganisms such as Candida albicans . Moreover, IL-12 production by splenic macrophages was diminished in CD40L(-/-) mice and interferon-gamma production by CD4 and CD8 T cells was decreased . CD4 T cells retained the ability to express a costimulatory molecule, CTLA-4, but showed a decrease in CD28 expression . This latter molecule is implicated in a positive effect on proliferation, cytokine production and survival of T cells . Collectively these data demonstrate that absence of CD40L correlates with (i) reduced antimicrobial activity of natural effector cells; (ii) reduction of the magnitude of T cell response; and (iii) increase of fungal growth in the brain . These findings suggest that disruption of CD40/CD40L may be deleterious for development of an efficient immune response to C . neoformans and may identify potential molecular targets for novel immunotherapeutic approaches FEMS Yeast Res, 2004 Jan, 4(4-5), 377 - 88 The internal transcribed spacers and 5.8S rRNA gene show extensive diversity among isolates of the Cryptococcus neoformans species complex; Katsu M et al.; Sequences of the internal transcribed spacer (ITS) region including the 5.8S rRNA gene delineated seven genotypes within the three varieties of Cryptococcus neoformans via specific combinations of eight nucleotide differences located at positions 10, 11, 15, 19, 108 (ITS1), 221 (5.8S), 298 and 346 (ITS2) . The ITS types correlated to polymerase chain reaction fingerprint/random amplification of polymorphic DNA (RAPD) molecular types: with ITS type 1 (ATACTAGC)=C . neoformans var . grubii, molecular types VNI+VNII and the serotype A allele of the AD hybrid, VNIIIA; ITS type 2 (ATATAGGC)=the serotype D allele of the AD hybrid, VNIIIB, and C . neoformans var . neoformans, VNIV; and ITS type 3 (GCGCTGGC) and ITS type 7 (ACGCTGGC)=VGI=RAPD type III, ITS type 4 (ACACTGAC)=VGII=RAPD type II, ITS type 5: (ACACTGGG)=VGIII=RAPD type I, ITS type 6 (ACACTGGC)=VGIV=RAPD type IV, all corresponding to C . neoformans var . gattii . Cloned sequences from serotype AD revealed that the hybrid serotype is diploid at the ITS1-5.8S-ITS2 locus carrying the ITS type 1 (ATACTAGC) and the ITS type 2 (ATATAGGC) alleles . ITS sequencing is a useful technique for genotyping the three C . neoformans varieties and for subtyping within C . neoformans var . gattii. FEMS Yeast Res, 2004 Jan, 4(4-5), 361 - 7 Cyclic AMP signaling in Cryptococcus neoformans; Pukkila-Worley R et al.; As pathogenic microorganisms establish an infection, they must be able to sense host-specific signals and respond by elaborating determinants that allow for survival in these hostile conditions . Pathogen cell surface proteins detect these signals and activate signal transduction cascades that ultimately alter gene expression resulting in an adaptive cellular response . Here we review the mechanisms by which a pathogenic fungus uses the highly conserved cAMP signal transduction pathway to regulate cellular differentiation as well as its virulence potential. Zhonghua Er Ke Za Zhi, 2003 Sep, 41(9), 692 - 6 {Molecular diagnosis of the specific DNA patterns of 16S-23S rRNA gene of bacteria}; Shang SQ et al.; OBJECTIVE: To establish the specific 16S-23S rRNA gene spacer regions pattern in different bacteria using polymerase chain reaction (PCR), restriction fragment length polymorphism (RFLP), DNA cloning and sequences analysis . METHODS: A pair of primers were selected from highly conserved sequences adjacent to the 16S-23S rRNA spacer region . Bacterial DNA of sixty-one strains of standard bacteria and corresponding clinical isolates representative of 20 genera and 27 species was amplified by PCR, and further studied by RFLP, DNA cloning and sequences analysis . Meanwhile, all specimens were examined by bacterial culturing and PCR-RFLP analysis . RESULTS: The 27 different standard strains showed one, two, three or more than three bands . The sensitivity of PCR reached 2.5 colony-forming unit (CFU), and there was no cross reaction to the human, fungal or viral genomic DNAs . Fifteen species could be distinguished immediately by PCR, while another 10 species were further identified by Hinf I or Alu I digestion . Klebsiella pneumoniae (Kp) and Enterococcus durans (Ed) could not be differentiated from each other by Alu I or Hinf I digestion . The spacer sequences of the Kp and Ed were 908 bp and 909 bp, respectively, and they differed only at the site of the 779th nucleotide . The former was G, and the latter was A . The 760 - 790 bp sequence of Kp was as follows: CGACTGCACCGCCTCCTAC / GGCCGCGTATTC . The 760 - 790 bp sequence of Ed was as follows: CGACTGCAC CGCCTCCTAC / AGCCGCGTATTC . Only one enzyme XmaIII, could discriminate the two . The cleaving site of XmaIII is C downward arrow GGCCG . Kp DNA was cleaved into 778 bp and 130 bp fragments, while E . durans was not . Of 42 specimens with suspected septicemia, 15 were positive (35.7%) on blood culture, and 27 on PCR (64.29%) . The positive rate of PCR was significantly higher than that of blood culture (P < 0.01) . Of the six CSF specimens, one was positive for Staphylococcus epidermidis (Se) on culture as well as by PCR, while two specimens which were negative on cultures were positive by PCR and were diagnosed as Se according to its DNA pattern . One specimen was culture-positive for Cryptococcus neoformans (Cn) but was negative by PCR . The other two specimens were negative by both PCR and culture . Fifteen blood samples from healthy children were negative by both blood culture and PCR . CONCLUSIONS: The method of detecting bacterial 16S-23S rRNA spacer regions using PCR-RFLP techniques was specific, sensitive, rapid and accurate in detecting pathogens in clinical bacterial infections. Nihon Kokyuki Gakkai Zasshi, 2003 Dec, 41(12), 874 - 7 {A case of pulmonary cryptococcosis with multiple nodular shadows under treatment of bronchiolitis obliterans with organizing pneumonia (BOOP)}; Tashiro N et al.; A 66 year-old man was introduced to our hospital because of multiple infiltrative pulmonary shadows on February, 2001 . We diagnosed bronchiolitis obliterans with organizing pneumonia (BOOP) from the clinical and bronchoalveolar lavage fluid (BALF) findings, and initiated oral steroid therapy . Since the abnormal chest shadows disappeared, the dose of steroid was decreased and maintained at 10 mg/day . In August 2001, multiple infiltrative shadows returned, and we therefore increased the steroid dose to 30 mg/day . The expanding infiltrative shadows were then joined by new multiple nodular shadows . The bronchioalveolar lavage fluid revealed small bodies of cryptococcus species . A positive result for anti-cryptococcus antigen was also obtained from the serum . We then diagnosed pulmonary cryptococcosis without meningitis . Therapy was started with anti-mycotic agents including amphotericin-B, flucytosine and fluconazole, which proved successful . This case of opportunistic cryptococcus infection in an immunocompromized patient, which responded to anti-mycotic therapy, is reported. J Natl Med Assoc, 2003 Dec, 95(12), 1211 - 5 Emerging opportunistic fungal infections: where are we heading? Idemyor V. Medical mycology involves the study of pathogenic fungi and their identification in the laboratory . Mycology has developed into a field that demands the attention of all clinicians treating patients in hospitals . Interest in medical mycology has grown in recent years due to a dramatic rise in the rates of fungal infections . An overview of well-known medically significant opportunistic fungi, such as Candida, Cryptococcus, Aspergillus and Zygomycetes, as well as emerging fungal pathogens, are discussed . Antifungal failures in these individuals are high; consequently, mortality rates are also high, despite standard therapy with amphotericin-B, lipid-associated formulation of amphotericin-B and the azoles . This underscores the need for new approaches and therapies to improve outcomes in high-risk individuals. Bull Soc Pathol Exot, 2003 Nov, 96(4), 291 - 4 {AIDS at Central University Hospital of Brazzaville: experience of the "Grands Enfants" pediatric department}; Loufoua-Lemay AM et al.; The epidemiological factors, clinical aspects and short term evolution of children infected by aids were assessed over a period of five years in "Grands Enfants" paediatric service of Brazzaville CHU . The medical reports of 81 patients have been collected, their average age was 9 years old +/- 3.5 . The HIV 1 was the only identified virus . 59% of children's parents were divorced, and 7% were single . The infection transmission was vertical in 70% of cases, transfusional in 28% of cases and indeterminate in 1% of cases . The symptoms were: the impairment of body status in 90% of cases, shown by a weight loss in 89% of cases; a long-term fever was observed in 56% of cases and a chronic diarrhoea was noticed in 54% of cases . The lymphadenopathies and digestive Candida were observed in 26% and 36% of cases . 6% of the patients had a psychomotor retardation . The respiratory infections and the diarrhoea were pathologies the most frequently noticed . The associated infections were essentially pulmonary tuberculosis 40% of cases, the otitis, the pneumococcal and cryptococcal meningia 2% of cases . The Burkitt lymphoma and the Kaposi sarcoma were shown in 1% of cases . The evolution was shown by a high mortality due to a dehydration in 59% of cases, a respiratory pathology in 22% of cases and anaemia in 15% of cases . This high mortality points out the problem of the treatment of patients, a problem increased by family poverty. Tidsskr Nor Laegeforen, 2004 Jan 8, 124(1), 28 - 30 {Cryptococcal meningitis}; Sivertsen EA et al.; BACKGROUND: Cryptococcus neoformans causes systemic disease in patients with immunodeficiency . The incidence of cryptococcal meningitis has increased in parallel with that of HIV infection . Cancer is also a known predisposing factor . MATERIAL AND METHODS: We present two case reports and a review of the literature concerning the epidemiology, diagnostics and treatment of cryptococcal meningitis . RESULTS: The incidence of cryptococcal meningitis in Scandinavia seems to be lower than in other parts of the world . Clinical signs and symptoms are often uncharacteristic . Detection of antigen in spinal fluid is a sensitive and fast test . INTERPRETATION: Cryptococcal meningitis is a rare disease, often with uncharacteristic symptoms . Patients with haematological malignancies have a higher risk of contracting this disease . It is a differential diagnosis when neurological symptoms occur in these patients. J Clin Microbiol, 2004 Jan, 42(1), 481 - 3 First report of a case of meningitis caused by Cryptococcus adeliensis in a patient with acute myeloid leukemia; Rimek D et al.; Cryptococcus adeliensis is a recently described new fungal species which has been isolated from decaying algae in Terre Adelie, Antarctica . We report the first known case of meningitis caused by C . adeliensis in a patient with acute myeloid leukemia undergoing allogeneic peripheral blood stem cell transplantation. J Clin Microbiol, 2004 Jan, 42(1), 404 - 7 Specific oligonucleotide primers for identification of Cladophialophora carrionii, a causative agent of chromoblastomycosis; Abliz P et al.; Cladophialophora carrionii is one of the relatively common causative agents of chromoblastomycosis . We have developed the specific oligonucleotide primer set based on the internal transcribed spacer regions of ribosomal DNA for the rapid identification of this pathogen . PCR with this primer set amplified a 362-bp amplicon from C . carrionii strains . From other relevant dematiaceous species, including medically important dematiaceous fungi, such as Fonsecaea pedrosoi, Phialophora verrucosa, and Exophiala dermatitidis, and eight species of medically important yeasts, such as Candida albicans and Cryptococcus neoformans var . neoformans, the primer set did not produce any amplicon . PCR with this primer set may be a useful tool for the identification of C . carrionii. J Clin Microbiol, 2004 Jan, 42(1), 380 - 3 Evaluation of the NCCLS M44-P disk diffusion method for determining susceptibilities of 276 clinical isolates of Cryptococcus neoformans to fluconazole; Pfaller MA et al.; We evaluated the NCCLS M44-P fluconazole disk diffusion method in comparison with the NCCLS M27-A2 broth microdilution method for determining the susceptibility of 276 isolates of Cryptococcus neoformans . Disk diffusion testing was performed using Mueller-Hinton agar supplemented with 2% glucose and 0.5 microg of methylene blue/ml . Among the 276 isolates, 259 (93.8%) were susceptible, 16 (5.8%) were susceptible--dose dependent, and 1 (0.4%) was resistant to fluconazole as determined by the NCCLS broth microdilution method . The overall categorical agreement between the two methods was 86%, with 0% very major errors, 2% major errors, and 12% minor errors . The disk diffusion method using Mueller-Hinton agar supplemented with glucose and methylene blue appears to be a useful approach for determining the fluconazole susceptibility of C . neoformans. J Clin Microbiol, 2004 Jan, 42(1), 236 - 8 Nonvalue of culturing cerebrospinal fluid for fungi; Barenfanger J et al.; No studies have evaluated the efficacy of culturing cerebrospinal fluid (CSF) for fungi . Because of the facts that the most common fungi responsible for meningitis grow well in media utilized for routine bacterial cultures and that cryptococcal antigen tests are commonly ordered, the efficacy of routinely performing fungal cultures specifically to recover fungi has been questioned . We examined data from 1225 samples of CSF which were cultured for both bacteria and fungi . Twelve specimens yielded fungi, 10 from fungal cultures and 8 from bacterial cultures .