Sante, 2000 Sep-Oct, 10(5), 329 - 37 {Spectrum of opportunistic infections in subjects infected with HIV at Libreville, Gabon}; Okome-Nkoumou M et al.; This retrospective study (1992-1996) was carried out in the Internal Medicine Department of the Jeanne Ebori Foundation at Libreville . We analyzed 351 files with the aim of counting the numbers of cases of various opportunistic infections and estimating the frequencies of these infections . The prevalence of seropositivity for HIV was 27.7% in the hospital population . Young adults from modest socioeconomic backgrounds were found to consult at stage IV of the infection . Oropharyngeal candidiasis (37%), zona (18.5%), salmonellosis (18.2%) and tuberculosis (14.5%) were the most frequently diagnosed opportunistic infections . Intestinal parasitoses, cryptococcosis, radiculomeningitis, cerebral toxoplasmosis and visceral fungal infections were diagnosed less frequently . The prevalence of Kaposi's sarcoma was 6.3% . Most of the opportunistic infections encountered were treatable but the mortality rate was high (11.7%) due to late diagnosis, the lack of availability of drugs and the poor economic conditions of the population. Mol Microbiol, 2001 Jan, 39(1), 166 - 75 Extracellular phospholipase activity is a virulence factor for Cryptococcus neoformans; Cox GM et al.; The human pathogenic fungus Cryptococcus neoformans secretes a phospholipase enzyme that demonstrates phospholipase B (PLB), lysophospholipase hydrolase and lysophospholipase transacylase activities . This enzyme has been postulated to be a cryptococcal virulence factor . We cloned a phospholipase-encoding gene (PLB1) from C . neoformans and constructed plb1 mutants using targeted gene disruption . All three enzyme activities were markedly reduced in the mutants compared with the wild-type parent . The plb1 strains did not have any defects in the known cryptococcal virulence phenotypes of growth at 37 degrees C, capsule formation, laccase activity and urease activity . The plb1 strains were reconstituted using the wild-type locus and this resulted in restoration of all extracellular PLB activities . In vivo testing demonstrated that the plb1 strain was significantly less virulent than the control strains in both the mouse inhalational model and the rabbit meningitis model . We also found that the plb1 strain exhibited a growth defect in a macrophage-like cell line . These data demonstrate that secretory phospholipase is a virulence factor for C . neoformans. Mol Microbiol, 2000 Dec, 38(5), 1017 - 26 Characterization of the MFalpha pheromone of the human fungal pathogen cryptococcus neoformans; Davidson RC et al.; Cryptococcus neoformans is an important human pathogenic fungus with a defined sexual cycle and well-developed molecular and genetic approaches . C . neoformans is predominantly haploid and has two mating types, MATa and MATalpha . Mating is known to be regulated by nutritional limitation and thought also to be regulated by pheromones . Previously, a portion of the MATalpha locus was cloned, and a presumptive pheromone gene, MFalpha1, was identified by its ability to induce conjugation tube-like filaments when introduced by transformation into MATa cells . Here, the ability of the MFalpha1 gene to induce these morphological changes in MATa cells was used as a phenotypic assay to perform a structure-function analysis of the gene . We show that the MFalpha1 open reading frame is required for the morphological response of MATa cells . We also find that the cysteine residue of the C-terminal CAAX motif is required for activity of the MFalpha1 pheromone . In addition, we use a reporter system to measure the expression levels of the MFalpha1 pheromone gene and find that two signals, nutrient starvation and the presence of factors secreted by mating partner cells, impinge on this promoter and regulate MFalpha1 expression . We identify a second pheromone gene, MFalpha2, and show phenotypically that this gene is also expressed . Finally, we have synthesized the MFalpha1 pheromone and show that only the predicted mature modified form of the alpha-factor peptide triggers morphological responses in MATa cells. Proc Natl Acad Sci U S A, 2000 Dec 19, 97(26), 14455 - 60 Identification of the MATa mating-type locus of Cryptococcus neoformans reveals a serotype A MATa strain thought to have been extinct; Lengeler KB et al.; Cryptococcus neoformans is an opportunistic fungal pathogen with a defined sexual cycle involving mating between haploid MATa and MATalpha cells . Here we describe the isolation of part of the MATa mating-type locus encoding a Ste20 kinase homolog, Ste20a . We show that the STE20a gene cosegregates with the MATa mating type in genetic crosses, maps within the mating-type locus on a 1.8-Mb chromosome, and is allelic with the MATalpha locus . We identify the first MATa isolate of the most common pathogenic variety of C . neoformans (serotype A, variety grubii) which had been thought to be extinct . This serotype A MATa strain is sterile, fails to produce mating pheromone, and is less virulent than a serotype A MATalpha strain in an animal model . Our studies illustrate an association of mating type with virulence and suggest that, like Candida albicans, pathogenic isolates of C . neoformans may be largely asexual. Infect Immun, 2001 Jan, 69(1), 213 - 20 Activation of Valpha14(+) natural killer T cells by alpha-galactosylceramide results in development of Th1 response and local host resistance in mice infected with Cryptococcus neoformans; Kawakami K et al.; We examined the effect of alpha-galactosylceramide (alpha-GalCer) on the synthesis of gamma interferon (IFN-gamma) and local resistance in mice infected intravenously with Cryptococcus neoformans . The level of IFN-gamma in serum increased on day 3, reached a peak level on day 7, and decreased to the basal level on day 14 postinfection in mice treated with alpha-GalCer, while in vehicle-treated mice, no increase was detected at any time points except for a small increase on day 7 . Such effects were not observed in NKT-KO mice . In CD4KO mice, minor synthesis of IFN-gamma was detected on day 3 in sera but was completely abolished by day 7 . The alpha-GalCer-induced IFN-gamma production on day 3 was partially reduced in mice depleted of NK cells by treatment with anti-asialo-GM(1) antibody (Ab) . Spleen cells obtained from infected and alpha-GalCer-treated mice on day 7 produced a large amount of IFN-gamma upon restimulation with live organisms, while only a marginal level of production was detected in splenocytes from infected and vehicle-treated mice . Such effects were abolished in CD4KO and NKT-KO mice . Finally, the fungal loads in the lungs and spleen on days 7 and 14 were significantly reduced in alpha-GalCer-treated mice compared to those in control mice . In NKT-KO mice, local resistance elicited by alpha-GalCer was completely abolished, although no obvious exacerbation of infection was detected . Furthermore, treatment with anti-IFN-gamma monoclonal Ab mostly abrogated the protective effect of this agent . Thus, our results indicated that activation of Valpha14(+) NKT cells resulted in an increased Th1 response and local resistance to C . neoformans through production of IFN-gamma. Infect Immun, 2001 Jan, 69(1), 115 - 22 Serotype AD strains of Cryptococcus neoformans are diploid or aneuploid and are heterozygous at the mating-type locus; Lengeler KB et al.; Cryptococcus neoformans is a pathogenic basidiomycete with a defined sexual cycle involving mating between haploid yeast cells with a transient diploid state . C . neoformans occurs in four predominant serotypes (A, B, C, and D), which represent different varieties or species . Rare clinical and environmental isolates with an unusual AD serotype have been reported and suggested to be diploid . We found by fluorescence-activated cell sorter analysis that serotype AD strains are aneuploid or diploid . PCR analysis with primers specific for serotype A or D alleles of the CNA1, CLA4, and GPA1 genes revealed that both alleles are often present in serotype AD strains . PCR analysis with primers specific for genes in the MATa or MATalpha mating-type loci revealed that serotype AD strains are heterozygous for the mating-type locus . Interestingly, in several serotype AD strains, the MATalpha locus was derived from the serotype D parent and the MATa locus was inherited from a serotype A parent that has been thought to be extinct . Basidiospores from a self-fertile serotype AD strain bearing the putative serotype A MATa locus showed a very low viability ( approximately 5%), and no fertile serotype A MATa strain could be recovered . Serotype AD strains were virulent in a murine model . Hybrid AD strains could readily be isolated following a laboratory cross between a serotype A strain and a serotype D strain . In summary, serotype AD strains of C . neoformans are unusual aneuploid or diploid strains that result from matings between serotype A and D strains . Self-fertile isolates fail to undergo normal meiosis because of genetic divergence . Our findings further suggest that serotype A MATa strains may exist in nature. FEMS Immunol Med Microbiol, 2000 Dec, 29(4), 329 - 32 The effects of Cryptococcus neoformans-secreted antigens on tumor necrosis factor-alpha-induced intercellular adhesion molecule-1 expression on human lung epithelial cells; Merkel GJ et al.; Since primary infection with Cryptococcus neoformans usually occurs in the lungs, and since pulmonary cryptococcosis involves interactions between yeasts and alveolar epithelial cells, we have begun to study the effects of C . neoformans and its secreted antigens (SA) on epithelial reactions potentially associated with localized inflammation . We report here that SAs from encapsulated and acapsular strains of C . neoformans caused significant reductions in tumor necrosis factor-alpha (TNF-alpha)-induced intercellular adhesion molecule-1 (ICAM-1) expression on A549 lung epithelial cells in culture . We also present evidence that the reduction in ICAM-1 expression was not associated with SA-induced shedding of this adhesion molecule. Biochim Biophys Acta, 2000 Dec 20, 1509(1-2), 103 - 10 Sequencing and heterologous expression in Saccharomyces cerevisiae of a Cryptococcus neoformans cDNA encoding a plasma membrane H(+)-ATPase; Gorgojo B et al.; A cDNA containing an open reading frame encoding a putative plasma membrane H(+)-ATPase in the human pathogenic basidiomycetous yeast Cryptococcus neoformans was cloned and sequenced by means of PCR and cDNA library hybridization . The cloned cDNA is 3475 bp in length, containing a 2994 bp open reading frame encoding a polypeptide of 997 amino acids . As in the case of another basidiomycetous fungus (Uromyces fabae), the deduced amino acid sequence of CnPMA1 was found to be more homologous to those of P-type H(+)-ATPases from higher plants than to those from ascomycetous fungi . In order to prove the sequenced cDNA corresponds to a H(+)-ATPase, it was expressed in Saccharomyces cerevisiae and found to functionally replace its own H(+)-ATPase . Kinetic studies of CnPMA1 compared to ScPMA1 show differences in V(max) values and H(+)-pumping in reconstituted vesicles . The pH optimum and K(m) values are similar in both enzymes. J Neurol Neurosurg Psychiatry, 2001 Jan, 70(1), 113 - 6 Encephalomyelitis due to Cryptococcus neoformans var gattii presenting as spinal tumour: case report and review of the literature; Grosse P et al.; A 24 year old immunocompetent German resident is described who developed multifocal encephalomyelitis due to infection with Cryptococcus neoformans var gatti, commonly considered a disease of tropical regions . In the light of current knowledge on the epidemiology of C neoformans var gatti and the travel history of the patient it is assumed that the infection was acquired outside Europe . As exclusive intramedullary involvement is an outstandingly rare manifestation in spinal cryptococcosis, the particular diagnostic procedure and the therapeutic strategies are discussed Trends Microbiol, 2000 Dec, 8(12), 547 - 53 How does Cryptococcus get its coat? Doering TL. During the past few decades, increasing attention has focused on pathogenic fungi both as fascinating research subjects and as the agents of serious illness in diverse patient populations . In particular, opportunistic fungi such as Cryptococcus neoformans command notice as the ranks of their immunocompromised victims grow . C . neoformans is unique among fungal pathogens for its major virulence factor, a complex polysaccharide capsule . In this article, our current understanding of the structure and biosynthesis of the capsule is reviewed, as are the many questions that remain to be answered about how Cryptococcus gets its coat. Vaccine, 2000 Nov 22, 19(7-8), 924 - 30 A novel ELISA for determination of polysaccharide specific immunoglobulins; Spoljar BH et al.; A novel ELISA using Nunc CovaLink microtiter plates has been developed for the determination of polysaccharide specific antibodies in mice sera . Glucuronoxylomannan (GXM), a major capsular polysaccharide of Cryptococcus neoformans serotype B, was immobilised on CovaLink plates by covalent linkage of CNBr activated hydroxyl groups of the sugar and free bnd NH(2) groups of the plates . The binding characteristics of GXM to CovaLink and to conventional polystyrene ELISA plate (Costar) were compared . The differences were observed in quality of standard curve with anti-GXM MoAb 2H1 (R(2) values were 0.9468 and 0.9872 for Costar and CovaLink plates, respectively) and in absorbance values of sera of immunised mice which were 2.5 times higher on CovaLink than on Costar plates . Negative control was low and having the same value on both the plates . Using the novel ELISA we have analysed the influence of immunomodulatory peptidoglycan monomer on humoral immune response to GXM in mice . The treatment with the conjugate of the immunomodulator and GXM resulted in the increase of GXM-specific IgGs in mice sera . Finally, the method has been successfully modified for the determination of dextran-specific antibodies in mice sera, indicating that the described procedure could be applied for other polysaccharides that have &z.sbnd;OH groups available for CNBr activation. J Infect Dis, 2001 Jan 15, 183(2), 294 - 302 Epub 2000 Dec 08. Immune mediators in cerebrospinal fluid during cryptococcosis are influenced by meningeal involvement and human immunodeficiency virus serostatus; Lortholary O et al.; Pro- and anti-inflammatory mediators (tumor necrosis factor {TNF}-alpha, interleukin {IL}-6, IL-8, IL-10, and soluble TNF receptor II {sTNFR} II) were measured in cerebrospinal fluid (CSF) before treatment (day 0), and after 2 weeks and 3 months of antifungal therapy in 51 human immunodeficiency virus (HIV)-positive and 7 HIV-negative patients with culture-confirmed cryptococcosis . On day 0, all mediator concentrations, except IL-10 in HIV-positive patients, were higher in patients with meningeal, rather than extrameningeal cryptococcosis or in control subjects (P<.05) . For meningitis patients, all mediator levels, except sTNFR II, were higher in HIV-negative than HIV-positive patients (P<.05) . Day 0 CSF IL-8 levels were higher in HIV-positive patients receiving antiretroviral therapy than in untreated persons (P<.02) . Day 0 sTNFR II levels were higher in HIV-positive survivors at 3 months, and elevated levels were sustained in HIV-positive patients with meningitis . Overall, these data support the idea that inflammatory responses are crucial to the eradication of cryptococcal infections in the central nervous system. Nihon Kokyuki Gakkai Zasshi, 2000 Sep, 38(9), 670 - 5 {A clinicopathological study of pulmonary cryptococcosis--chest CT and pathologic correlation}; Kishi K et al.; We reviewed the clinicopathological features in 12 patients (7 males and 5 females; mean age 54 yr) with pulmonary cryptococcosis . Eleven of the patients were asymptomatic and the disease was detected by chest radiograph abnormalities . The underlying systemic disease had been diagnosed as diabetes mellitus in two . Chest CT scans showed a solitary nodule in 9 of the 12 patients, multiple nodules in 2, and infiltration in 1 . The nodular diameter was less than 2 cm in 10 of the 12 . All nodules were located in the subpleural region . On the chest CT, cavitary nodules, scattered nodules, or both, and spiculated nodules were difficult to distinguish from pulmonary tuberculosis and primary lung cancer, respectively . According to McDonnell's pathological classification of pulmonary cryptococcosis, the resected 8 lungs revealed peripheral pulmonary granuloma in 5 and granulomatous pneumonia in 3 . It is important to perform a pathological examination for the diagnosis of pulmonary cryptococcosis to avoid misdiagnosis as lung cancer or pulmonary tuberculosis. Rev Med Interne, 2000 Nov, 21(11), 955 - 60 {Cryptococcus neoformans infection in hematologic malignancies}; Vigouroux S et al.; PURPOSE: Cryptococcus is an opportunistic infection that affects immunodepressed patients and is a classical complication of AIDS-stage HIV infection . The aim of this study was to investigate Cryptococcus neoformans infections in patients with hematological malignancies . METHODS: Six cases have been described of cryptococcosis detected in Nantes, France over the past 10 years in patients with hematological malignancies . RESULTS: This infection has been found particularly in the context of lymphoproliferative disorders (chronic lymphocytic leukemia, Waldenstrom macroglobulinemia, Hodgkin's disease, and non-Hodgkin's lymphoma), and also following cytotoxic therapy . In four cases, the patients were treated with fludarabine, which rapidly caused long-duration marked lymphocytopenia, notably in CD4 cells . Cell-mediated immunity plays a major role in systemic defense against C . neoformans . It therefore seems that fludarabine favors the spread of cryptococcal infections . CONCLUSION: In the context of lymphoproliferative syndromes treated with cytotoxic drugs, in particular fludarabine, it appears important to take into account the possible presence of cryptococcal infection in the presence of respiratory, neurological or cutaneous disorders, so that a correct diagnosis can be made and the appropriate treatment administered. Int Microbiol, 1998 Sep, 1(3), 205 - 8 Yeast communities associated with sugarcane in Campos, Rio de Janeiro, Brazil; de Azeredo LA et al.; Yeast communities associated with sugarcane leaves, stems and rhizosphere during different phases of plant development were studied near Campos, in Rio de Janeiro, Brazil . Atmospheric temperature, soil granulometry and pH, and sugar cane juice degree Brix and pH were determined . Yeast communities associated with sugarcane were obtained after cellular extraction by shaking, blending and shaking plus sonication, and cultured on Yeast Nitrogen Base Agar plus glucose (0.5%) and Yeast Extract-Malt Extract Agar . No significant differences in yeast counts were found among the cellular extraction treatments and culture media . 230 yeast cultures were identified according to standard methods, and distinct yeast communities were found for each substrate studied . The prevalent species isolated from sugarcane were Cryptococcus laurentii, Cryptococcus albidus, Rhodotorula mucilaginosa and Debaryomyces hansenii. Braz J Infect Dis, 1997 Apr, 1(2), 60 - 67 Cryptococcal Meningitis; Graybill JR; Meningitis is the most common manifestation of disseminated cryptococcosis . Cryptococcal meningitis is the most common systemic mycotic infection in AIDS, which is the most frequent predisposing condition . The most severe complication is acute cerebral hypertension . Medical options for therapy have broadened considerably, but generally include initial intensive induction treatment followed, in patients with AIDS, by chronic suppression . With aggressive management of cryptococcal meningitis, mortality may be reduced to 10% or less. Microbiol Mol Biol Rev, 2000 Dec, 64(4), 746 - 85 Signal transduction cascades regulating fungal development and virulence; Lengeler KB et al.; Cellular differentiation, mating, and filamentous growth are regulated in many fungi by environmental and nutritional signals . For example, in response to nitrogen limitation, diploid cells of the yeast Saccharomyces cerevisiae undergo a dimorphic transition to filamentous growth referred to as pseudohyphal differentiation . Yeast filamentous growth is regulated, in part, by two conserved signal transduction cascades: a mitogen-activated protein kinase cascade and a G-protein regulated cyclic AMP signaling pathway . Related signaling cascades play an analogous role in regulating mating and virulence in the plant fungal pathogen Ustilago maydis and the human fungal pathogens Cryptococcus neoformans and Candida albicans . We review here studies on the signaling cascades that regulate development of these and other fungi . This analysis illustrates both how the model yeast S . cerevisiae can serve as a paradigm for signaling in other organisms and also how studies in other fungi provide insights into conserved signaling pathways that operate in many divergent organisms. Braz J Infect Dis, 1997 Oct, 1(5), 260 - 265 Bone Paracoccidioidomycosis in an HIV-Positive Patient; Miranda Aires E et al.; AIDS patients are vulnerable to infection by opportunistic microbes, including various fungi such as Pneumocystis carinii, Cryptococcus neoformans, Histoplasma capsulatum, Candida albicans and many others . However, the association of AIDS and infection with Paracoccidioides brasiliensis has been rarely recorded . We report a case of an HIV-positive patient with bone infection by this fungus with a clinical form not previously published . This clinical presentation included primarily a massive bone lesion, but it did not include the lymphatic and disseminated disease described in HIV-positive patients . The patient responded well to medical and surgical treatment . We suggest that patients with moderate, rather than severe, immunosuppression may have forms of paracoccidioidomycosis with a pathologic process intermediate to those seen in the immunologically normal host and the full AIDS syndrome. J Antimicrob Chemother, 2000 Dec, 46(6), 997 - 1000 Comparison of the Etest and microdilution method for antifungal susceptibility testing of Cryptococcus neoformans to four antifungal agents; Aller AI et al.; We performed a prospective study to compare the Etest and the microdilution method (NCCLS guidelines) for determining the MICs of fluconazole, itraconazole, flucytosine and amphotericin B for 35 strains of Cryptococcus neoformans . For the microdilution method (MDM) RPMI 1640 medium with 2% glucose was used for fluconazole, itraconazole and flucytosine, and Antibiotic Medium 3 for amphotericin B . For the Etest, RPMI 1640 medium with 2% glucose and solidified with 1.5% agar was used for the four antifungal agents . Amphotericin B was also tested on Antibiotic Medium 3 solidified with 1.5% agar . Fluconazole and flucytosine MICs by the Etest showed good correlation with the broth MDM (81.1 and 89.2% agreement within two dilutions, respectively) . With the tested population of itraconazole- and amphotericin B-susceptible isolates, the MIC agreement for itraconazole was 54%; amphotericin B showed the lowest agreement (8.1% on Antibiotic Medium 3 and 13.5% on RPMI). Mycoses, 2000, 43 Suppl 1, 61 - 8 {Fourier transform infrared spectroscopy, molecular biologic methods and antimyocotic susceptibility patterns for identification and differentiation of cryptococcus species}; Schmalreck AF et al.; Molecular biological methods as well as the FTIR method allows the rapid, reliable and reproducible determination and identification of Cryptococcus species from human, veterinary and environmental origin and their serovars . The results obtained by FTIR could be verified by the molecular methods . In addition, with the PCR and FTIR fingerprinting methods it is possible to distinctly group the serovars and differentiate the different Cryptococcus strains. Mycoses, 2000, 43 Suppl 1, 48 - 60 {Cryptococcus species--etiological agents of zoonoses or sapronosis?}; Blaschke-Hellmessen R; Cryptococcus strains are doubtless the cause of a sapronosis which occurs worldwide as cryptococcosis in humans and warm-blooded animals . The etiological agent is Cryptococcus neoformans with the varieties neoformans and gattii . The infection proceeds from environmental sources and not from animals suffering from cryptococcosis . Therefore the designation as zooanthroponosis is not right . There is a correlation between the geographical distribution of the varieties of Cryptococcus neoformans in the environments and the clinical manifestation of cryptococcosis . The reservoir of var . neoformans, serotypes A and D, are worldwide pigeons and pet birds which are frequent healthy carriers . They excrete these yeasts with their excrements . Bird droppings favour the propagation of cryptococci in natural substrates . The cryptococcal infection arises from inhalation of contaminated dust . Variety gattii, serotype B, occurs mainly in tropical and subtropical climates and its environmental niche has been identified in Eucalyptus trees . Var . gattii was detected in the air and in decaying wood debris under the canopies of these trees . Serotype C was isolated once from almond trees in Colombia . Cryptococcoses in men caused by var . gattii are endemic in Australia, California, Brazil and other tropical and subtropical regions . Modern molecular typing procedures are available for studying ecological problems of cryptococci and epidemiological questions of cryptococcosis . Immunosuppressed individuals, especially HIV infected persons and AIDS patients, should avoid contact with pigeons and pet birds. Mycoses, 2000, 43 Suppl 1, 36 - 9 {Host-pathogen relations in yeasts}; Gedek BR; Allogenic and autogenic factors are decisive for a colonization of warmblooded hosts by yeasts where they can live inside as symbiont, commensal or pathogen . To develop pathogenicity a capacity of yeasts for multiplication inside of humans or homoiothermic animals and resistance to environmental influences is not sufficient . Pathogenicity rather depends on the ability of structural morphological changes, phenomena of adherence and of microbial competition as well as the presence of genes for down-regulation of the host defense and for enzyme production enabling a parasitic life style . Among the yeasts Cryptococcus neoformans and some Candida species meet these requirements. Mycoses, 2000, 43 Suppl 1, 23 - 8 {Evidence of Cryptococcus neoformans in domestic and sports pigeons in Thyringia}; Kielstein P et al.; 19 strains of Cryptococcus neoformans var . neoformans were isolated from 17 (= 40.5%) of 42 investigated pigeon breeder flocks in Thuringia. Eur Radiol, 2000, 10(11), 1782 - 91 Correlation between high-resolution computed tomographic, magnetic resonance and pathological findings in cases with non-cancerous but suspicious lung nodules; Li F et al.; Computed tomography scans, including thin-section high-resolution computed tomography (HRCT), occasionally fail to differentiate between small non-cancerous nodules from lung cancers . We describe nine such lesions ( < 20 mm in diameter) initially identified through our screening program for lung cancer using CT scanning . Pathological diagnoses included nodular fibrosis (n = 4), granuloma (n = 1), cryptococcoma (n = 1), localised organising pneumonia (n = 1), inflammatory pseudo-tumour (n = 1) and sclerosing haemangioma (n = 1) . High-resolution CT findings, together with MRI findings with contrast-enhanced dynamic studies, were retrospectively evaluated . Additional cases should be identified and radiologically characterised in order to reduce the number of non-cancerous tumours that are treated by unnecessary surgery. Curr Infect Dis Rep, 2000 Aug, 2(4), 352 - 357 Cryptococcal Meningitis in HIV-Infected Patients; Powderly WG; Cryptococcosis is a fungal infection that is potentially deadly for and common among AIDS patients, in the United States and worldwide . Subacute meningitis and meningoencephalitis are typical, clinically . This article will review relevant aspects of cryptococcal meningitis in AIDS, focusing on the most recent information pertaining to pathogenesis, epidemiology, clinical syndromes, and treatment of this devastating disease. Curr Infect Dis Rep, 1999 Jun, 1(2), 160 - 165 Chronic Meningitis; Tunkel AR; Chronic meningitis may be caused by a large number of infectious agents, including spirochetes (Treponema pallidum, Borrelia burgdorferi), Mycobacterium tuberculosis, and fungi (primarily Cryptococcus neoformans) . The incidence of these specific causes of chronic meningitis has been impacted since the advent of HIV infection, and new information is also available on how this epidemic has affected populations in developing countries of the world . In the area of diagnostics, the development of polymerase chain reaction has been a major advance that has increased our capabilities for identifying etiologic agents (such as M . tuberculosis) that are difficult to culture . Finally, the management of patients with chronic meningitis has evolved, and the availability of new antifungal agents and adjunctive strategies has changed the approach to the patient with cryptococcal meningitis. Med Mycol, 2000 Oct, 38(5), 385 - 90 Environmental isolation of Cryptococcus neoformans var . gattii and C . neoformans var . neoformans in the city of São Paulo, Brazil; Montenegro H et al.; In order to determine the environmental occurrence of both varieties of Cryptococcus neoformans in the city of Sao Paulo, pigeon droppings and vegetable material from trees of the genus Eucalyptus were collected at typically crowded places . A total of 38 sites downtown where large heaps of pigeon droppings could be found were selected for sampling . Pigeon droppings from 10 (26.3%) of these sites were positive for C . neoformans var . neoformans in at least one sample . Twelve eucalyptus woods located within four municipal parks were also surveyed; vegetable material from Eucalyptus spp . trees were collected monthly over a 2-year period . C . neoformans var . gattii was recovered from a wood in Ibirapuera Park during the same season on two different occasions (November 1996 and November 1997); this park contained specimens of Eucalyptus camaldulensis, a natural habitat of C . n . var . gattii . C . n . var . neoformans was detected in a wood in Ibirapuera Park and Aclimacao Park . The results show that both C . n . var . neoformans and C . n . var . gattii are present in the urban environment of Sao Paulo city at sites where large numbers of people normally gather. Med Mycol, 2000 Oct, 38(5), 379 - 83 Possible primary ecological niche of Cryptococcus neoformans; Lazera MS et al.; To study hollows of living trees as natural habitats of Cryptococcus neoformans in an endemic area of cryptococcosis in the northeastern region of Brazil, samples of decaying wood were collected inside 32 hollows of living trees and plated on niger seed agar . Identification of C . neoformans was based upon morphological and physiological tests . Canavanine-glycine-bromothymol medium was used to screen the varieties and Crypto Check Iatron Kit to serotype the isolates . A total of 123 C . neoformans colonies were recovered from samples of six (18.5%) out of 32 hollow trees . C . neoformans var . neoformans and C . neoformans var . gattii were found occurring alone (pink shower tree, fig tree and pottery tree) or sharing the same hollow (pink shower tree) . Long lasting positivity (19-36 months) and significant number of cfu of C . neoformans per gram of decaying wood (0.15-21.7 x 10(3) cfu g(-1)) inside hollows of pink shower tree, fig tree and pottery tree were observed, indicating colonization of these habitats by the fungus . For the first time, C . n . var . neoformans and C . n . var . gattii were found sharing the same natural biotope, thus establishing a possible link between them in their life cycle in nature and suggesting the primary natural niche for the species. Med Mycol, 2000 Oct, 38(5), 343 - 53 Differing requirement for inducible nitric oxide synthase activity in clearance of primary and secondary Cryptococcus neoformans infection; Aguirre KM et al.; The role of nitric oxide in resistance to cryptococcal infection was investigated . Mice deficient in inducible nitric oxide synthase (INOS) did not survive a primary intratracheal infection as did INOS-replete control mice . Despite adequate recruitment of host cells and generation of interferon (IFN)-gamma and tumor necrosis factor (TNF)-alpha at the site of infection, INOS-deficient mice failed to clear yeast from their lungs by five weeks of infection, in contrast to wild-type mice . INOS-deficient mice also had higher yeast brain burdens than did control mice after a primary intracerebral infection . Therefore, generation of nitric oxide is required for resistance to primary cryptococcal infection . However, INOS-deficient mice vaccinated subcutaneously and rechallenged intravenously had lung and brain yeast burdens equivalent to those of vaccinated controls, and therefore expressed effective acquired immunity to Cryptococcus neoformans . Cells harvested from infected INOS-deficient mice by bronchoalveolar lavage acted as anti-cryptococcal effectors in vitro at an effector:target ratio of 100:1, provided IFN-gamma was present, but did not inhibit yeast proliferation at a 10:1 effector:target ratio as cells from wild-type mice did . Therefore, INOS activity is important for anti-cryptococcal function of effectors of immunity during the primary response, but not for the generation or expression of secondary immunity to C . neoformans. Arch Pharm (Weinheim), 2000 Oct, 333(10), 347 - 54 Synthesis and in vitro antifungal and cytotoxicity evaluation of thiazolo-4H-1,2,4-triazoles and 1,2,3-thiadiazolo-4H-1,2,4-triazoles; Jalilian AR et al.; The increasing clinical importance of drug-resistant fungal pathogens has lent additional urgency to microbiological and antifungal research . Various thiazolo(or 1,2,3-thiadiazolo)thiosemicarbazides (2a-2e), 3-thiono-1,4-dihydrotriazolothiazoles-(or 1,2,3-thiadiazoles) (3a-3e), their related substituted thio-4H-1,2,4-triazoles (4a-4p) and sulfones (5a-5o) were synthesized . Most of the compounds tested for antifungal activity exhibited significant effects against Cryptococcus neoofrmans and Sacchromyces cerevisiae at MIC ranges of 0.53 to 12.5 micrograms/mL, whereas their activities were moderate against Candida albicans and weak against Aspergillus fumigatus . At 10 ppm concentration, all compounds showed low toxicity on brine shrimps (higher than 80% survival), except compounds 4c and 2c . At 100 ppm concentration most of the compounds showed toxicity except compounds 2b, 2e, 3c, 3d, 3e, and 4e . Compounds 4b, 4c, and 4h showed in vitro cytotoxicity against Kbalb cell lines and compounds 4c and 4g against 143B cell lines at 0.1 mM concentration. AIDS, 2000 Oct 20, 14(15), 2349 - 54 Improving survival following AIDS in Australia, 1991-1996 . National HIV Surveillance Committee; Li Y et al.; OBJECTIVE: To describe the pattern of survival following AIDS . DESIGN: National surveillance for AIDS diagnoses . METHODS: AIDS cases in adults/adolescents (aged 13 years or older at AIDS diagnosis) and deaths following AIDS were notified to the national HIV surveillance centre by the diagnosing doctor through State/Territory health authorities . The date of last medical contact for each case living with AIDS was updated annually . RESULTS: By 30 June 1999, 4814 AIDS cases, diagnosed in Australia in 1991-1996, and 3193 deaths following AIDS had been notified to the National AIDS Registry . Median survival following AIDS was 17.7 months . Survival following AIDS increased from 16.0 months in 1991 to 27.7 months in 1996 . Factors independently associated with improved survival were year of AIDS diagnosis, late HIV diagnosis, CD4+ cell count greater than 50 x 10(6) cells/l, age of less than 45 years and presentation with Pneumocystis carinii pneumonia only or Kaposi's sarcoma only . The risk of death declined over time when the initial AIDS-defining illness was Pneumocystis carinii pneumonia only {adjusted hazard ratio (AHR) = 0.91, P < 0.0005}; other opportunistic infections (AHR, 0.88; P < 0.0005); Kaposi's sarcoma only (AHR, 0.92; P = 0.025); and central nervous system conditions (HIV encephalopathy, cryptococcosis, toxoplasmosis) (AHR, 0.92; P = 0.012) . No time trend was observed for survival following diagnoses of non-Hodgkin's lymphoma or other multiple illnesses . CONCLUSIONS: Survival following AIDS has improved in Australia, especially among cases diagnosed in 1995 and 1996 . Temporal improvements in survival following AIDS were coincident with the introduction of combination antiretroviral treatment for HIV infection and suggest that treatment is effective in limiting disease progression among people with advanced HIV infection. J Infect Dis, 2001 Jan 1, 183(1), 51 - 8 Epub 2000 Nov 21. Interleukin-12 counterbalances the deleterious effect of human immunodeficiency virus type 1 envelope glycoprotein gp120 on the immune response to Cryptococcus neoformans; Pietrella D et al.; The mechanism involved in the envelope glycoprotein gp120-induced Th2 response to Cryptococcus neoformans was investigated . Peripheral blood mononuclear cells (PBMC) from healthy donors were treated with human immunodeficiency virus gp120 and an encapsulated or acapsular strain of C . neoformans in the presence or absence of glucuronoxylomannan, the major capsular polysaccharide . gp120 inhibited early and late production of interleukin (IL)-12 by PBMC . This reduction paralleled IL-10 induction and inhibited translocation of CD40 to the surface of monocytes . Flow cytometric analysis revealed that gp120 down-regulated the expression of IL-12 receptor beta2 subunit on T cells responding to C . neoformans . Because the IL-12/IL-12 receptor beta2 subunit pathway is critical for the Th1 differentiation process, underexpression demonstrates that gp120 contributes to Th2 bias . Exogenous IL-12 added simultaneously with gp120 up-regulated interferon-gamma secretion and limited IL-4 production . These results suggest that gp120 limits the Th1 response to C . neoformans and that exogenous IL-12 could offset this effect. Mycopathologia, 1999, 148(1), 1 - 7 The Cryptococcus neoformans genome sequencing project; Heitman J et al.; Cryptococcus neoformans is a basidiomycete that can cause life-threatening meningoencephalitis in patients with and without impaired immune function . Cryptococcosis is usually an opportunistic infection in patients with compromised immunity as a consequence of HIV-1 infection, steroid administration, cancer chemotherapy, sarcoidosis, diabetes, or inherited immune system defects . This pathogenic yeast has a defined sexual cycle, which allows classical genetic analysis . Molecular biology approaches, including transformation and gene disruption by homologous recombination, and animal models for studies of virulence are both well developed . Recently an international consortium convened to begin the C . neoformans genome sequencing project, and we review here background and arguments for this project . We also discuss the importance of this project to the biology and virulence of this organism in particular, and to virulence in general. J Immunol, 2000 Dec 1, 165(11), 6429 - 36 The role of macrophage inflammatory protein-1 alpha/CCL3 in regulation of T cell-mediated immunity to Cryptococcus neoformans infection; Olszewski MA et al.; Macrophage inflammatory protein-1alpha (MIP-1alpha/CCL3) is a CC chemokine required for optimal recruitment of leukocytes in response to cryptococcal Ags . MIP-1alpha is expressed in the lungs by day 6 post Cryptococcus neoformans infection and could play a role in the development of cell-mediated immunity . To address this possibility, wild-type (MIP-1alpha(+/+)) mice and MIP-1alpha knockout (MIP-1alpha(-/-)) mice were infected intratracheally with a highly virulent strain of C . neoformans (145A) . MIP-1alpha message was detected in the lungs on days 3, 7, and 14 in MIP-1alpha(+/+) mice, but it was undetectable in MIP-1alpha(-/-) mice . On day 16, MIP-1alpha(-/-) mice had a 7-fold increase in C . neoformans burden in the lungs, but no decrease in pulmonary leukocyte recruitment . MIP-1alpha(+/+) and MIP-1alpha(-/-) mice had similar numbers of recruited lymphocytes and monocytes/macrophages . Notably, MIP-1alpha(-/-) mice had a significantly greater number of eosinophils . MIP-1alpha(-/-) mice had extremely high levels of serum IgE . This switch of immune response to a T(2) phenotype was associated with enhanced IL-4 and IL-13 expression in the lungs of MIP-1alpha(-/-) mice compared with MIP-1alpha (+/+) mice . Progression of pulmonary cryptococcosis in the presence of nonprotective T(2) immunity resulted in profound lung damage in MIP-1alpha(-/-) mice (eosinophilic crystal deposition, destruction of lung parenchyma, and pulmonary hemorrhage) . Twelve-week survival was dramatically decreased in MIP-1alpha(-/-) mice . These studies, together with our previous studies, demonstrate that MIP-1alpha plays a role in both the afferent (T(1)/T(2) development) and efferent (T(1)-mediated leukocyte recruitment) phases of cell-mediated immunity to C . neoformans. Infect Immun, 2000 Dec, 68(12), 7049 - 60 Human antibodies against a purified glucosylceramide from Cryptococcus neoformans inhibit cell budding and fungal growth; Rodrigues ML et al.; A major ceramide monohexoside (CMH) was purified from lipidic extracts of Cryptococcus neoformans . This molecule was analyzed by high-performance thin-layer chromatography (HPTLC), gas chromatography coupled with mass spectrometry, and fast atom bombardment-mass spectrometry . The cryptococcal CMH is a beta-glucosylceramide, with the carbohydrate residue attached to 9-methyl-4,8-sphingadienine in amidic linkage to 2-hydroxyoctadecanoic acid . Sera from patients with cryptococcosis and a few other mycoses reacted with the cryptococcal CMH . Specific antibodies were purified from patients' sera by immunoadsorption on the purified glycolipid followed by protein G affinity chromatography . The purified antibodies to CMH (mainly immunoglobulin G1) bound to different strains and serological types of C . neoformans, as shown by flow cytofluorimetry and immunofluorescence labeling . Transmission electron microscopy of yeasts labeled with immunogold-antibodies to CMH and immunostaining of isolated cell wall lipid extracts separated by HPTLC showed that the cryptococcal CMH predominantly localizes to the fungal cell wall . Confocal microscopy revealed that the beta-glucosylceramide accumulates mostly at the budding sites of dividing cells with a more disperse distribution at the cell surface of nondividing cells . The increased density of sphingolipid molecules seems to correlate with thickening of the cell wall, hence with its biosynthesis . The addition of human antibodies to CMH to cryptococcal cultures of both acapsular and encapsulated strains of C . neoformans inhibited cell budding and cell growth . This process was complement-independent and reversible upon removal of the antibodies . The present data suggest that the cryptococcal beta-glucosylceramide is a fungal antigen that plays a role on the cell wall synthesis and yeast budding and that antibodies raised against this component are inhibitory in vitro. Eur J Med Res, 2000 Oct 30, 5(10), 424 - 30 HIV-related nontuberculous mycobacterial infection: incidence, survival analysis and associated risk factors; Arasteh KN et al.; To evaluate the incidence and survival time for AIDS-patients affected by different stages of nontuberculous mycobacterial (NTM) infection we performed a retrospective study . Data of 1540 hospitalised AIDS-patients was analyzed with respect to survival time and incidence rates . The overall incidence rate of NTM following AIDS was 16.6/100 person-years (PY), with an increase from 12.1/100PY (1987-1990) to 18.9/100PY (1991-1994) . Antiretroviral therapy (ART) and toxoplasmosis prophylaxis reduced the risk of NTM disease whereas CD4 cells <40/ microl at time of the first AIDS defining illness led to a 2.5 fold higher risk . Pneumocystis carinii pneumonia (PCP), wasting syndrome and PCP prophylaxis increased the risk of progression from colonization to dissemination . Cryptococcus neoformans infection, wasting syndrome, PCP prophylaxis and CD4 cells <40/ microl were linked to immediate NTM dissemination . Though the incidence of NTM dissemination increased by the factor 1.56 in 1991-1994, survival did not differ between patients with and without NTM infection. Clin Infect Dis, 2000 Nov, 31(5), 1303 - 6 Persistence of cryptococcomas on neuroimaging; Hospenthal DR et al.; Three previously normal patients with cryptococcal meningitis had intracranial lesions on computed tomography and magnetic resonance imaging that persisted for >5 years after successful cure with antifungal drugs . Persistence of lesions on neuroimaging should not be misinterpreted as evidence of active cryptococcosis. J Infect Dis, 2000 Dec, 182(6), 1791 - 5 Epub 2000 Nov 08. The effect of the echinocandin analogue caspofungin on cell wall glucan synthesis by Cryptococcus neoformans; Feldmesser M et al.; The echinocandin derivative caspofungin (MK-0991, L-743,872) inhibits 1,3-beta-d-glucan synthesis and is active against several medically important fungi but is relatively ineffective against Cryptococcus neoformans . To investigate the mechanism of C . neoformans resistance, the prevalence of 1,3- and 1,6-beta-d-glucan linkages was determined in cells grown with and without caspofungin, using affinity-purified antisera and gold particle immunoelectron microscopy . Cryptococcal strains ATCC 24067 (serotype D) and MY2061 (serotype A) were studied . Growth at 4 microg/mL of caspofungin reduced both glucan linkages in both strains . However, growth at 2 microg/mL resulted in reduced 1,6-beta-d-glucan linkage only for MY2061 . Inhibition of 1,6-beta-d-glucan synthesis may be an additional mechanism of action for pneumocandins . The relatively low efficacy of caspofungin against C . neoformans may result from reduced activity against C . neoformans glucan synthase or from yet undiscovered mechanisms of action operative in other fungal pathogens but not in C . neoformans. Kekkaku, 2000 Sep, 75(9), 557 - 60 {Mechanisms of pathogenicity and host defense in infections by intracellular parasitic microbes}; Mitsuyama M et al.; Mycobacterium tuberculosis is one of the intracellular parasitic bacteria escaping the intracellular killing inside macrophages . The aim of this symposium was to get some insight into the mechanism of pathogenicity and host defense in M . tuberculosis infection, which has not yet been elucidated well, by the presentation of up-to-date knowledge on these aspect in infection with different intracellular parasitic microbes . Dr . Yoshikai (Nagoya Univ.) indicated that TLR is involved in the initial response of host against S . choleraesuis . Among the cytokines contributing to the induction of specific immunity, the importance of IL-15 was emphasized, based on their own experimental data using IL-15 transgenic mice and the application of anti-IL-15 antibody in vivo . Dr . Yoshida (Kyushu Univ.) reviewed the mechanisms of intracellular growth of Legionellae . Several genes so far identified as essential genes in intra-macrophage growth appeared to be similar to those encoding type 3 secretion system observed in Shigellae . There is a significant strain difference in the growth of L . pneumophila inside macrophages and such difference seemed to be under the control of a gene at chromosome 13, Lgn 1 . The investigation of difference in the mode of escape among various Legionella . spp . may provide a novel mechansim in bacterial invasion and escape . Dr . Kawamura (Kyoto Univ.) summarized some new reports on the molecular mechanism of the inhibition of P-L fusion by M . tuberculosis . He emphasized the importance of the alteration in phagosomal maturation as indicated by the accumulation of TACO protein . The possible involvement of TLR in the recognition of Mycobacterial cells and its LAM was discussed . Dr . Kawakami (Ryukyu Univ.) first discussed the possibility that Cryptococcus neoformans, a fungal pathogen, could be regarded as one of the intracellular parasitic microbes . His presentation mainly focused on the TH1-Th2 balance in the expression of host defense against C . neoformans in mice . From their experimental infection using attenuated strain TC-13 in various cytokine-knock out mice, the pivotal role of both IL-12 and IL-18 was clearly indicated. Arch Med Res, 2000 Jul-Aug, 31(4), 393 - 8 The clinical spectrum of neurological manifestations in AIDS patients in Mexico; Gongora-Rivera F et al.; BACKGROUND: Neurological complications may be present clinically in up to 39% of patients infected with HIV or AIDS . Some reports have shown different profiles of neurological illness related to geographic variations and the population studied . METHODS: This retrospective study describes the neurological manifestations of patients with AIDS seen between 1990 and 1998 at a single neurological referral hospital in Mexico City . RESULTS: One hundred forty-nine patients were included, 133 males (89%) and 16 females (10.7%) . The average age was 33.8 years (9 to 75 years) . Upon admission, only 50 patients (33.6%) were known to be seropositive to HIV-1 . In 75 patients (50.3%), the neurological illness was definitory of AIDS and also was its first recognized clinical manifestation . The most common infection problems were brain toxoplasmosis (32.2%), meningeal cryptococcosis (21.5%), tuberculosis (8.7%), and AIDS-dementia complex (8.7%) . There were eight (5.4%) cases of ischemic cerebrovascular disease and four (2 . 7%) neoplasms . Two primary brain lymphomas and single cases of astrocytoma and oligodendroglioma, progressive multifocal leukoencephalopathy (PML), aseptic meningitis, acute encephalitis, transverse myelitis, myopathy, and cranial neuropathy were also seen . CONCLUSION: In comparison with other studies of neurological complications of AIDS, opportunistic infections amenable to treatment in our population were more common . A high case fatality rate was observed, as was a large proportion of patients in whom the neurological illness was the first manifestation of HIV infection or AIDS due to denied, unknown, or unrecognized risk factors for HIV infection. Nippon Ishinkin Gakkai Zasshi, 2000, 41(4), 241 - 4 {Mycological and serological diagnosis of cryptococcosis}; Ikeda R et al.; Methods for the diagnosis of cryptococcosis have been established, including serotyping and serodiagnosis . Slide agglutination tests with factor sera, the phenol oxidase test, and the growth test at 37C are used for rapid identification of Cryptococcus neoformans . We identified 140 strains and found that 86, 10, and 4% of the isolates were serotypes A, D, and A-D, respectively . Twelve of 14 serotype D strains were isolated from cutaneous cryptococcosis . The most reliable method of serodiagnosis is the latex agglutination (LA) test for detection of polysaccharide antigen combined with protease pretreatment . The LA test is also used for prognosis . The clearance of cryptococcal polysaccharide antigen often takes a few years after treatment . To model the persistence of cryptococcal polysaccharides, we examined the clearance of antigen from the blood of rabbits injected with polysaccharide . The distribution and elimination half-lives of the antigen suggest the prolonged survival of C . neoformans . Recently, the number of cases of C . albidus and C . laurentii has been increasing . The antigenic pattern and the sensitivity of C . albidus in the LA test are the same as that of C . neoformans serotype A . In contrast, C . laurentii does not react with factor sera for C . neoformans and the reactivity with sensitized latex is extremely low . These results were supported by the chemical structures of polysaccharides from these species . We should consider non-neoformans cases in both the identification of isolates and in serodiagnosis. Nippon Ishinkin Gakkai Zasshi, 2000, 41(4), 221 - 8 {New developments in therapy of deep mycoses}; Yamaguchi H; Over the past two decades the incidence of deep mycoses caused by several major groups of fungal pathogens such as Candida spp., aspergilli, Cryptococcus neoformans and zygomycetes has risen steadily . Moreover, opportunistic fungal infections due to Fusarium spp., Trichosporon spp., Pseudallescheria boydii and other emerging pathogens, as well as fluconazole-resistant Candida albicans, all of which are often resistant to existing antifungal drugs, are also encountered more and more frequently . This makes it more difficult for the clinician to achieve successful treatment . Thus there is an urgent need to develop new antifungal agents or formulations with advantages over and/or complimentary to existing drugs . This review focuses on current approaches to antifungal chemotherapy with special reference to the clinical development of new drugs, including (ii) lipid formulations of amphotericin B, (i) second-generation azoles and (iii) antifungal lipopeptides. J Neuroimmunol, 2000 Nov 1, 111(1-2), 10 - 4 Cryptococcal glucuronoxylomannan delays translocation of leukocytes across the blood-brain barrier in an animal model of acute bacterial meningitis; Lipovsky MM et al.; In bacterial meningitis, neurological damage is associated with a high influx of polymorphonuclear leukocytes (PMN) into the brain . Previous data suggest that the capsular component of the fungus C . neoformans, glucuronoxylomannan (GXM), interferes with PMN-migration into the cerebrospinal fluid (CSF) . Therefore, a rabbit model of bacterial meningitis was treated intravenously with GXM . This resulted in (1) a reduction of PMN in the CSF at 6 h (P=0.05), (2) reduced peak TNF-alpha concentrations in the CSF, and (3) diminished tissue inflammation and intravascular margination of PMN in GXM-treated animals . Thus, GXM may represent a novel adjuvant anti-inflammatory agent in bacterial meningitis. Singapore Med J, 2000 Feb, 41(2), 80 - 2 Cryptococcal meningitis resulting in irreversible visual impairment in AIDS patients--a report of two cases; Ng CW et al.; Cryptococcus neoformans is the leading cause of meningitis in patients with Acquired Immune Deficiency Syndrome (AIDS) and is associated with high mortality rate . Presenting symptoms include fever, nausea and vomiting, altered mentation, headache and meningismus . Cryptococcal meningitis is not infrequently complicated by raised intracranial pressure and visual sequelae (sometimes by blindness) . In patients who survive the infection, the most debilitating outcome appears to be visual impairment or blindness . Management of impending visual complication combines medical and surgical treatment modalities . We report two cases of cryptococcal meningitis associated with visual impairment. Can Vet J, 2000 Oct, 41(10), 799 - 800 A mixed fungal infection in a dog: sporotrichosis and cryptococcosis; Shany M; Unusual ulcerated masses protruding from both nostrils of a 3-year-old terrier were diagnosed histologically as sporotrichosis, and regressed with iodide therapy . Cryptococcus neoformans was recovered from new lesions that appeared near the dog's eye and on the extremities . All lesions regressed with itraconazole therapy. J Antimicrob Chemother, 2000 Nov, 46(5), 815 - 8 Susceptibility of Cryptococcus neoformans by the NCCLS microdilution and Etest methods using five defined media; Petrou MA et al.; The susceptibility of 12 isolates of Cryptococcus neoformans to amphotericin B, 5-fluorocytosine, fluconazole, itraconazole and ketoconazole was tested using the NCCLS and Etest methods with yeast nitrogen base (YNB) pH 5.6 and pH 7.0, RPMI MOPS pH 7.0 with and without added glucose (2%) and RPMI buffered with phosphate buffer to pH 7.0 . Some isolates yielded poor growth in RPMI MOPS after 72 h . Tests indicated that YNB pH 5.6 was the best medium for 5-fluorocytosine but was unsuitable for ketoconazole . In conclusion, YNB pH 7.0 or RPMI MOPS with 2% glucose can be used with either method. J Clin Microbiol, 2000 Nov, 38(11), 4021 - 5 Serological relationships of Cryptococcus spp.: distribution of antigenic factors in Cryptococcus and intraspecies diversity; Ikeda R et al.; The antigenic formulas of 34 species in the genus Cryptococcus were determined by using type strains and eight factor sera prepared from adsorption experiments with Cryptococcus neoformans serotypes . These antigenic factors were shared by 19 species . The strains used could be divided into eight serological groups . The patterns of groups 1, 2, 3, 5, and 6 were the same as the patterns of C . neoformans serotypes A, D, A-D, B, and C, respectively . The species belonging to group 4 reacted to factor sera 1, 2, and 3 . Group 7 contained one species that reacted only to factor serum 1 . The 15 species in group 8 did not react to any of the factor sera used . Compared to the reported molecular phylogenetic tree, the serological and phylogenetic data were correlated in the Filobasidium lineage . All the members of the albidus clade in the Filobasidium lineage had antigens 1, 2, and 3, and all the strains in the magnus clade belonged to serogroup 8 . Moreover, intraspecies diversity was examined using strains of C . curvatus, C . humicolus, and C . laurentii . Serological heterogeneity was observed in the species C . humicolus and C . laurentii, as well as in phylogenetic relationships previously published . Using serological features, similarities and differences between Cryptococcus species were demonstrated . Our study contributes to a better description of the genus Cryptococcus and related species phenotypically and phylogenetically. Yeast, 2000 Nov, 16(15), 1397 - 403 Characterization of the L41 gene in Cryptococcus neoformans: its application as a selectable transformation marker for cycloheximide resistance; Varma A et al.; A transformation system using resistance to the antibiotic cycloheximide as a dominant selectable marker was developed for the pathogenic yeast Cryptococcus neoformans . A 3.5 kb DNA fragment containing a gene encoding the ribosomal protein L41 was cloned from a wild-type strain of C . neoformans which is sensitive to cycloheximide . The open reading frame of the L41 gene contains five introns and encodes a protein of 107 amino acids, which is similar to those reported for other yeasts . The cycloheximide resistance gene to be used as a marker was constructed by replacing a DNA segment of the wild-type L41 gene, which contained the amino acid proline at its 56th position with a homologous DNA segment from a mutant strain resistant to cycloheximide that contained leucine in that position . Cycloheximide resistant transformants were obtained by electroporation on YEPD plates, supplemented with 10-20 microg/ml cycloheximide, at a maximum efficiency of 300 transformants/microg plasmid DNA . While with other genes, most transformants of serotype D in C . neoformans maintain the transforming DNA as episomes, the cycloheximide-resistant transformants were all the result of ectopic genomic integration events . Rinsho Byori, 2000 Jul, 48(7), 575 - 9 {Current HIV therapy and its clinical problems}; Oka S; HIV-specific protease inhibitors(PI) have been available in Japan since 1997 . Since then, highly active anti-retroviral therapy(HAART) including two reverse transcriptase inhibitors combined with PI became the main strategy of HIV treatment . After introducing HAART, incidence of most opportunistic infections dramatically decreased, resulted a steep decline of AIDS death in Japan as well as in the United States . However, several unexpected problems related to HAART have been coming up . One is a lipodystrophy syndrome(LDS) which is a novel side effect caused by PI . Lipid disposition was noted associated with hyperlipidemia and/or hyperglycemia . Ischemic heart diseases will emerge in patients with LDS in future . Another one is inflammatory reactions to some opportunistic pathogens, such as Mycobacteria, Pneumocystis carinii, cryptococcus, and so on, occurred during course of immune reconstitution after HAART . This reaction is sometimes too severe to continue HAART and corticosteroid is often required to control the reaction . How to diagnose and how to manage the reaction are to be determined in future. Semin Neurol, 2000, 20(3), 307 - 22 Fungal meningitis; Gottfredsson M et al.; Fungi provide many benefits to humans . However, some of these fungi have the ability to become human pathogens . All the major fungal pathogens can produce meningitis . From the common cryptococcal meningitis to the rare fungal meningitis caused by a dimorphic or filamentous fungus, medical issues are discussed in this review on a fungus-specific basis . Both primary (Cryptococcus, Blastomyces, Histoplasma, Coccidioides, and other dimorphic fungi) and secondary (Aspergillus, Candida, and a series of molds) fungal pathogens can produce life-threatening central nervous system infections . These infections require immediate and precise diagnosis and carefully selected management strategies to optimize outcomes . In this review, we examine the epidemiology, pathogenesis, clinical manifestations, and treatment for fungal meningitis in all the major fungal groups. Mol Ecol, 2000 Oct, 9(10), 1471 - 81 Multiple gene genealogies reveal recent dispersion and hybridization in the human pathogenic fungus Cryptococcus neoformans; Xu J et al.; Cryptococcus neoformans (= Filobasidiella neoformans) is a significant emerging fungal pathogen of humans . To understand the evolution of this pathogen, 34 strains were obtained from various locations around the world and fragments of four genes were sequenced from each . These strains represented all three varieties and five serotypes . The four sequenced genes are: (i) the mitochondrial large ribosomal subunit RNA; (ii) the internal transcribed spacer region of the nuclear rRNA, including ITS1, 5.8S rRNA subunit and ITS2; (iii) orotidine monophosphate pyrophosphorylase; and (iv) diphenol oxidase . Phylogenetic analyses indicated considerable divergence among lineages, which corresponded to the current classification of C . neoformans into three varieties . However, there is no apparent phylogeographic pattern . Significant incongruences were observed among gene genealogies . The analyses indicated that the major lineages in C . neoformans diverged tens of millions of years ago but have undergone recent dispersion and hybridization. Br J Ophthalmol, 2000 Nov, 84(11), 1275 - 81 Postmortem histological survey of the ocular lesions in a British population of AIDS patients; Pecorella I et al.; AIMS: To study ocular pathology and systemic correlations in a series of 73 postmortem eyes from British patients who died from AIDS before the introduction of a HAART regimen . METHODS: The eyes were studied with conventional histology, special histochemical stainings, and immunohistochemistry . RESULTS: 72.6% of the cases showed chronic uveal inflammation, caused by opportunistic agents in 37.7% of them (cytomegalovirus (CMV) in 30.1%, C neoformans in 5.6%, and Gram positive bacteria in 1.8%) . Cytoid bodies were noted in 10/73 eyes, three linked to CMV retinitis . Six retinal haemorrhages, four of which were secondary to CMV, were found . 14 specimens (19 . 1%) showed foci of calcification, and a further 11 (15%) calcium oxalate deposits . In no cases were the calcific deposits suspected clinically . Six eyes (8.2%) did not show any abnormality . CONCLUSIONS: CMV retinitis is the most common (28.7%, 21/73) ocular infection in this series and may occur either during or in the absence of systemic dissemination . Conversely, ocular cryptococcosis appears to be an epiphenomenon of systemic and CNS disease . No other opportunistic ocular infections were present in this series . Interesting findings were the presence of intraocular precipitates of calcium oxalate and calcium phosphate or carbonate in a significant number of cases (15% and 19%, respectively), and the high prevalence of idiopathic uveal inflammation (43.8%). J Pharmacol Exp Ther, 2000 Nov, 295(2), 655 - 61 Identification of R146225 as a novel, orally active inhibitor of interleukin-5 biosynthesis; Van Wauwe J et al.; Interleukin (IL)-5 regulates the growth, differentiation, and activation of eosinophils . When activated, eosinophils release an array of proinflammatory and cytotoxic products and act as prominent effector cells in the process of allergic inflammation . Depriving eosinophils of IL-5 may therefore represent a viable approach to treat allergic disorders . This study describes the identification of R146225, a novel six-substituted azauracil derivative, as a potent, orally active inhibitor of IL-5 biosynthesis, capable of reducing pulmonary eosinophilia in mice . In vitro, R146225 inhibited IL-5 protein formation by activated human whole blood (IC(50) = 34 nM), human peripheral blood mononuclear cells (IC(50) = 24 nM), and murine spleen cells (IC(50) = 6 nM) . In contrast, the compound enhanced generation of interferon-gamma and had little or no inhibitory effect on the production of IL-2 and IL-4 . Reverse transcription-polymerase chain reaction analysis of stimulated whole blood cells indicated R146225's ability to down-regulate IL-5 mRNA expression . In vivo p.o . administration of R146225 (2.5 mg/kg) to mice before an i.v . anti-CD3 antibody challenge reduced IL-5 but enhanced interferon-gamma serum levels, without affecting IL-2 and IL-4 production . Analogous to the in vitro results, R146225 suppressed splenic IL-5 mRNA expression, while message levels of the other cytokines remained unchanged . Moreover, p.o . dosing of R146225 (0.6-2.5 mg/kg) dose dependently reduced the pulmonary accumulation of eosinophils induced in mice by an intranasal instillation of Cryptococcus neoformans . Based on these data, R146225 may be useful in the therapy of eosinophil-driven allergic conditions. Chem Pharm Bull (Tokyo), 2000 Oct, 48(10), 1422 - 6 A new pentanorlanostane derivative, cladosporide A, as a characteristic antifungal agent against Aspergillus fumigatus, isolated from Cladosporium sp; Hosoe T et al.; In the course of searching for new antifungal agents, a new pentanorlanostane derivative, cladosporide A (1), was isolated along with ergosterol, ergosterol peroxide and 23,24,25,26,27-pentanorlanost-8-ene-3beta,22-diol (2) from Cladosporium sp . as a characteristic antifungal agent against the human pathogenic filamentous fungus Aspergillus fumigatus . The structure of 1 was established as 3beta,22-dihydroxy-23,24,25,26,27-pentanorlanostane-29-al by spectroscopic and chemical investigation and X-ray crystallographic analysis . Inhibitory activity against A . fumigatus (IC80 0.5-4.0 microg/ml) was observed for cladosporide A (1), but no activity was observed against pathogenic yeasts, Candida albicans and Cryptococcus neoformans, and other pathogenic filamentous fungi, Aspergillus niger and A . flavus . The 4beta-aldehyde residue in 1 might be essential for the antifungal activity, since 23,24,25,26,27-pentanorlanost-8-ene-3beta,22-diol (2) showed no inhibition against the above four fungi. Trop Med Int Health, 2000 Oct, 5(10), 687 - 91 AIDS caused by HIV1 and HIV2 infection: are there clinical differences? Results of AIDS surveillance 1986-97 at Fann Hospital in Dakar, Senegal; Ndour M et al.; OBJECTIVE: To compare the clinical manifestations observed in AIDS patients infected with HIV2 and HIV1 infection . METHODS: The medical records of AIDS patients hospitalized between January 1986 and July 1997 at the Department of Infectious Diseases of Fann Hospital, Dakar, were reviewed . RESULTS: 599 hospitalizations (76%) were HIV1 seropositive patients, 137 (17%) were HIV2 seropositive patients and 54 (7%) were patients serologically dually reactive to HIV1 and HIV2 . There was no significant difference in medium CD4 lymphocyte count between patients with HIV1 and HIV2 infection . Chronic diarrhoea and diarrhoea caused by bacterial infections were more frequently observed in HIV2-infected individuals . Oral candidiasis and chronic fever were more often noted in patients with HIV1 infection . Bacterial and cryptococcal meningitis was only observed among patients with HIV1 infection . CONCLUSIONS: Certain clinical differences were observed comparing AIDS patients with HIV1 and those with HIV2 infection . As there is no clear physiopathological explanation for these differences, additional studies with larger numbers of AIDS patients are needed to determine whether these differences are real. J Infect, 2000 Jul, 41(1), 92 - 4 Bone marrow cryptococcal infection in the acquired immunodeficiency syndrome; Pantanowitz L et al.; OBJECTIVES: To describe the bone marrow lesions in eight cases of Cryptococcus neoformans infection involving the bone marrow in HIV-infected patients . METHODS: Archival bone marrow biopsies from patients with HIV-related cryptococcosis of the bone marrow were retrospectively reviewed . Cryptocococcal organisms were identified on haematoxylin- and eosin-stained slides and confirmed using mucicarmine staining . RESULTS: Yeast cells stimulated a granulomatous response in all cases despite immunosuppression . The number of cryptococcal organisms appeared to be inversely proportional to the adequacy of the granulomatous response . All patients had a cytopenia . CONCLUSIONS: The ability to mount a tissue response in order to localize organisms is retained in patients with AIDS . Infection of the bone marrow with cryptococci may act in synergy with HIV to cause cytopenia. J Food Prot, 2000 Oct, 63(10), 1359 - 68 Antimicrobial effect of rosemary extracts; Del Campo J et al.; A rosemary extract commercially exploited (Oxy'less) as an antioxidant of lipids in foods was dissolved in ethanol (100 mg/ml), and the solution was tested against foodborne microorganisms . For gram-positive bacteria, the MIC of the ethanolic solution was 1% for Leuconostoc mesenteroides, 0.5% for Listeria monocytogenes, 0.5% for Staphylococcus aureus, 0.13% for Streptococcus mutans, and 0.06% for Bacillus cereus . It slowed the growth of Penicillium roquefortii and Botrytis cinerea . Up to 1% of the ethanolic solution had no activity on the gram-negative bacteria Escherichia coli, Salmonella Enteritidis, and Erwinia carotovora and on the yeasts Rhodotorula glutinis and Cryptococcus laurentii . Antibacterial activity of the rosemary extract was strongly influenced by the composition of the media . The MIC was reduced by low pH, high NaCl contents, and low temperatures . Low pH and high NaCl concentration had a synergistic effect on the MIC of the rosemary extract for S . aureus . Lipids, surface-active agents, and some proteins decreased its antibacterial activity, whereas pectin had no effect . The inhibitory effect was little modified by heat treatment (100 degrees C) . The natural microflora of pasteurized zucchini broth was inhibited by 0.5% of the rosemary extract . The antibacterial activity was linked to the compounds extracted with hexane, which are presumably phenolic diterpenoids. Bioorg Khim, 2000 Aug, 26(8), 613 - 6 {Production of beta-xylosidase by the yeast Cryptococcus podzolicus}; Shubakov AA; In studying the beta-xylosidase production by yeast Cryptococcus podzolicus (Basidiomycetes), it was shown to be an inducible secretory enzyme . Xylooligosaccharides generated from xylan and methyl beta-xyloside were found to induce the beta-xylosidase synthesis . The enzyme activity in the medium containing xylan or methyl beta-xyloside was 1.0 and 1.5 U/ml, respectively; this production level is similar to that achievable at the beta-xylosidase production by mycelial fungi. Mycopathologia, 1999, 147(3), 121 - 4 Cryptococcus neoformans in bird excreta in the city zoo of Cali, Colombia; Caicedo LD et al.; The presence of Cryptococcus neoformans was studied in bird excreta and in the air circulating in and around bird cages in the City Zoo of Cali, Colombia, between August 1994 and April 1995, using a sunflower seed agar culture medium for fungus isolation . A total of 380 samples was studied, 110 from droppings and 270 from Petri dishes placed inside (148) and outside (122) the cages . C . neoformans var neoformans was found in only two cases, one from bird excreta (0.9%) and the other from air inside a cage (0.7%) . The former positive sample was collected from the cracks of a dead tree where two crested caracaras (Polyborus plancus) roosted; the feces were dry, accumulated, and with a pH of 6 . The other positive sample was found inside the cage of these birds; however, samples taken in a dispersion study at 0.5, 1, 5 and 10 m around this cage were all negative . It appears that this low isolation rate is due to adequate cleaning and disinfection procedures used in the city zoo of Cali. Indian J Med Res, 2000 Aug, 112, 56 - 60 Changing scenario of cryptococcosis in a tertiary care hospital in north India; Chakrabarti A et al.; BACKGROUND & OBJECTIVES: With the increase in the number of patients of AIDS, the incidence of cryptococcosis is on the rise in India . It was therefore considered important to evaluate the predisposing factors, laboratory investigations and outcome of patients with cryptococcosis in this changed scenario . METHODS: We assessed 58 patients with cryptococcosis retrospectively over a five year period (January 1995-December 1999) at the Nehru Hospital, Postgraduate Institute of Medical Education and Research (PGIMER), Chandigarh . RESULTS: The annual incidence of cryptococcosis in PGIMER, Chandigarh has increased about 15 fold from 1970-1982 (pre AIDS era) to 1995-1999 (present series) . Of the 47 patients studied for predisposing factors, 36 patients were identified with predisposing factors, HIV infection (57.4%) was the commonest followed by haematologic malignancies (6.3%) and renal transplant (4.2%) . Forty one patients were diagnosed by isolation of the organism as well as antigen detection in cerebrospinal fluid/serum, 9 by isolation alone and 8 by antigen detection alone . Quantitative antigen titres were done in 38 patients and a significantly higher (P < 0.01) antigen titre (> 512) was observed in HIV positive patients as compared to HIV negative patients . All isolates tested were of Cryptococcous neoformans var neoformans biotype and no resistance to antifungal agents was noted . Twenty of 41 patients receiving treatment improved . The results were compared with other studies available from India . INTERPRETATION & CONCLUSION: The incidence of cryptococcosis is on the rise in this part of north India and this can be attributed to an increase in AIDS cases. Mycoses, 2000 Sep, 43(7-8), 299 - 301 Anti-Cryptococcus activity of combination of extracts of Cassia alata and Ocimum sanctum; Ranganathan S et al.; The paper reports the anti-Cryptococcus activity of combination of ethanolic extracts of leaves of Cassia alata and Ocimum sanctum . The activity of combination of the extracts was heat-stable and worked at acidic pH. Antimicrob Agents Chemother, 2000 Nov, 44(11), 3180 - 3 Activities of an intravenous formulation of itraconazole in experimental disseminated Aspergillus, Candida, and Cryptococcus infections; Odds FC et al.; An intravenous (i.v.) formulation of itraconazole was evaluated in disseminated fungal infection models in guinea pigs . In acute disseminated Candida albicans and Aspergillus fumigatus infections, treatment at 5 mg/kg of body weight twice a day (b.i.d.) significantly prolonged survival . In these models and in animals with chronic disseminated cryptococcosis, itraconazole given i.v . at 2.5 and 5 mg/kg b.i.d . greatly reduced the proportions of organs with culture-detectable fungal burdens . The efficacy of i.v . itraconazole in these animal models justifies its further investigation for the treatment of life-threatening mycoses in humans. Infect Immun, 2000 Nov, 68(11), 6257 - 64 Human neutrophil-mediated nonoxidative antifungal activity against Cryptococcus neoformans; Mambula SS et al.; It has long been appreciated that polymorphonuclear leukocytes (PMN) kill Cryptococcus neoformans, at least in part via generation of fungicidal oxidants . The aim of this study was to examine the contribution of nonoxidative mechanisms to the inhibition and killing of C . neoformans . Treatment of human PMN with inhibitors and scavengers of respiratory burst oxidants only partially reversed anticryptococcal activity, suggesting that both oxidative and nonoxidative mechanisms were operative . To define the mediators of nonoxidative anticryptococcal activity, PMN were fractionated into cytoplasmic, primary (azurophil) granule, and secondary (specific) granule fractions . Incubation of C . neoformans with these fractions for 18 h resulted in percent inhibition of growth of 67.4 +/- 3.4, 84.6 +/- 4.4, and 29.2 +/- 10.5 (mean +/- standard error, n = 3), respectively . Anticryptococcal activity of the cytoplasmic fraction was abrogated by zinc and depletion of calprotectin . Antifungal activity of the primary granules was significantly reduced by pronase treatment, boiling, high ionic strength, and magnesium but not calcium . Fractionation of the primary granules by reverse phase high-pressure liquid chromatography on a C(4) column over an acetonitrile gradient revealed multiple peaks with anticryptococcal activity . Of these, peaks 1 and 6 had substantial fungistatic and fungicidal activity . Peak 1 was identified by acid-urea polyacrylamide gel electrophoresis (PAGE) and mass spectroscopy as human neutrophil proteins (defensins) 1 to 3 . Analysis of peak 6 by sodium dodecyl sulfate-PAGE revealed multiple bands . Thus, human PMN have nonoxidative anticryptococcal activity residing principally in their cytoplasmic and primary granule fractions . Calprotectin mediates the cytoplasmic activity, whereas multiple proteins, including defensins, are responsible for activity of the primary granules. Infect Immun, 2000 Nov, 68(11), 6196 - 201 The Cryptococcus neoformans gene DHA1 encodes an antigen that elicits a delayed-type hypersensitivity reaction in immune mice; Mandel MA et al.; When mice are vaccinated with a culture filtrate from Cryptococcus neoformans (CneF), they mount a protective cell-mediated immune response as detected by dermal delayed-type hypersensitivity (DTH) to CneF . We have identified a gene (DHA1) whose product accounts at least in part for the DTH reactivity . Using an acapsular mutant (Cap-67) of C . neoformans strain B3501, we prepared a culture filtrate (CneF-Cap67) similar to that used for preparing the commonly used skin test antigen made with C . neoformans 184A (CneF-184A) . CneF-Cap67 elicited DTH in mice immunized with CneF-184A . Deglycosylation of CneF-Cap67 did not diminish its DTH activity . Furthermore, size separation by either chromatography or differential centrifugation identified the major DTH activity of CneF-Cap67 to be present in fractions that contained proteins of approximately 19 to 20 kDa . Using N-terminal and internal amino acid sequences derived from the 20-kDa band, oligonucleotide primers were designed, two of which produced a 776-bp amplimer by reverse transcription-PCR (RT-PCR) using RNA from Cap-67 to prepare cDNA for the template . The amplimer was used as a probe to isolate clones containing the full-length DHA1 gene from a phage genomic library prepared from strain B3501 . The full-length cDNA was obtained by 5' rapid amplification of cDNA ends and RT-PCR . Analysis of DHA1 revealed a similarity between the deduced open reading frame and that of a developmentally regulated gene from Lentinus edodes (shiitake mushroom) associated with fruiting-body formation . Also, the gene product contained several amino acid sequences identical to those determined biochemically from the purified 20-kDa peptide encoded by DHA1 . Recombinant DHA1 protein expressed in Escherichia coli was shown to elicit DTH reactions similar to those elicited by CneF-Cap67 in mice immunized against C . neoformans . Thus, DHA1 is the first gene to be cloned from C . neoformans whose product has been shown to possess immunologic activity. Infect Immun, 2000 Nov, 68(11), 6147 - 53 Phagocytosis and protein processing are required for presentation of Cryptococcus neoformans mitogen to T lymphocytes; Syme RM et al.; In addition to eliciting antigen specific T-cell-mediated immunity, Cryptococcus neoformans possesses a mitogen (CnM) that activates naive T cells to proliferate . This mechanism of T-cell activation is accessory cell dependent and major histocompatibility complex unrestricted . CnM-induced T-cell proliferation correlates with internalization of the organism, suggesting that intracellular processing is required to liberate CnM prior to presentation to T cells . To determine whether phagocytosis and processing are required, various inhibitors of accessory cell uptake and processing were used . C . neoformans was observed within the accessory cells . Paraformaldehyde fixation of the accessory cell abrogated presentation of CnM to T cells, indicating that a dynamic accessory cell surface was required . A lysosomotropic agent abrogated the response to CnM but had no effect on a control stimulus that did not require processing . Both aspartic acid and cysteine protease inhibitors blocked effective processing of CnM, so that it was unable to stimulate T cells . Finally, an inhibitor of microfilament polymerization abrogated proliferation to CnM . These results indicate that the mitogenic activity of C . neoformans requires phagocytosis of the organism, lysosomal or endosomal processing, proteolytic activity, and microfilament polymerization and intracellular transport as a prerequisite for T-cell proliferation. Diagn Microbiol Infect Dis, 2000 Oct, 38(2), 87 - 93 A comparison of dynamic characteristics of fluconazole, itraconazole, and amphotericin B against Cryptococcus neoformans using time-kill methodology; Burgess DS et al.; This study evaluated the in vitro pharmacodynamics of fluconazole, itraconazole, and amphotericin B against Cryptococcus neoformans . MICs were determined for three clinical isolates according to NCCLS guidelines (M27) . Time-kill studies were performed using antifungal concentrations of 0.25-32 x MIC and inocula of 10(3) and 10(5) CFU/ml . At predetermined time points over 72 hours, samples of each inoculum/drug combination were withdrawn and plated using a spiral plater . Colony counts were determined after incubation at 35 degrees C for 48 hours . Area under the kill curves (AUKCs) were plotted versus the AUC/MIC ratios . Inoculum effect was evaluated by calculating an estimated AUKC for the low inoculum then comparing it to the measured low inoculum using the unpaired Student's t-test . The MICs of fluconazole and itraconazole for isolate 97-1199, 97-1061, and 97-585 were 2, 4, 32 microg/ml and 0.03, 0.06, 0 . 5 microg/ml, respectively . For amphotericin B, the MIC was 0 . 25 microg/ml for each isolate . The triazoles demonstrated fungistatic activity against each isolate at both inocula with the exception of itraconazole against C . neoformans 97-585 . Maximal suppression was noted at concentrations 8-16 x MIC correlating with an AUC/MIC of 192 for both inocula . Conversely, amphotericin B was fungicidal and displayed concentration-dependent activity against each isolate at both inocula . Maximal killing was observed at concentrations >4 x MIC for the low inoculum and >8 x MIC for the high inoculum for each isolate . No statistically significant differences were detected between the measured and estimated AUKCs for each antifungal agent . In conclusion, our results suggest that the triazoles were most effective against C . neoformans when concentrations were maintained at 8-16 x MIC . Amphotericin B, on the other hand, was concentration-dependent; thus, greater activity was exerted at higher concentrations. J Bacteriol, 2000 Nov, 182(21), 6222 - 7 Mapping of the Cryptococcus neoformans MATalpha locus: presence of mating type-specific mitogen-activated protein kinase cascade homologs; Karos M et al.; In this study we investigated the relationship between the MATalpha locus of Cryptococcus neoformans and several MATalpha-specific mitogen-activated protein (MAP) kinase signal transduction cascade genes, including STE12alpha, STE11alpha, and STE20alpha . To resolve the location of the genes, we screened a cosmid library of the MATalpha strain B-4500 (JEC21), which was chosen for the C . neoformans genome project . We isolated several overlapping cosmids spanning a region of about 71 kb covering the entire MATalpha locus . It was found that STE12alpha, STE11alpha, and STE20alpha are imbedded within the locus rather than closely linked to the locus . Furthermore, three copies of MFalpha, the mating type alpha-pheromone gene, a MATalpha-specific myosin gene, and a pheromone receptor (CPRalpha) were identified within the locus . We created a physical map, based on the restriction enzyme BamHI, and identified both borders of the MATalpha locus . The MATalpha locus of C . neoformans is approximately 50 kb in size and is one of the largest mating type loci reported among fungi with a one-locus, two-allele mating system. Diagn Microbiol Infect Dis, 2000 Sep, 38(1), 29 - 36 Commercial systems for fluconazole susceptibility testing of yeasts: comparison with the broth microdilution method; Posteraro B et al.; Fluconazole susceptibility was tested in 100 clinical yeast isolates (65 Candida albicans, 13 C . glabrata, 8 C . tropicalis, 7 C . parapsilosis, 3 Saccharomyces cerevisiae, 1 each of C . krusei, C . lusitaniae, Cryptococcus neoformans, Rhodotorula glutinis) and two control strains (Candida krusei ATCC 6258, C . parapsilosis ATCC 22019) using broth microdilution (reference method), disk diffusion, Etest strips, Sensititre YeastOne, Candifast, Integral System Yeasts . Using M27-A breakpoints, isolates were classified as susceptible (81%), susceptible-dose dependent or Resistant with broth dilution . Rates of concordance with the reference method were good for Sensititre YeastOne, Etest and disc-diffusion (81.2%-94.7%) but very low for the Candifast (3.1%) and Integral System (16.6%), which classified most susceptible isolates as resistant . Lack of standardisation (inoculum, medium composition) and non-objective interpretation schemes may be the cause of their poor performance . Sensititre YeastOne, Etest and disc-diffusion are potentially useful for fluconazole antifungal susceptibility testing of yeasts in clinical laboratories. J Ethnopharmacol, 2000 Nov, 73(1-2), 191 - 8 Antifungal activity of extracts from medicinal plants used by First Nations Peoples of eastern Canada; Jones NP et al.; From literature describing medicinal usage of plants by First Nations Peoples in eastern Canada, 18 eastern Canadian plants were selected and tested for their antifungal activities . Eight randomly selected tropical plants were also tested for comparative purposes . Four groups of plants were obtained: popular antimicrobial-remedy (n=6), popular non-antimicrobial-remedy (n=6), random temperate (n=6) and random tropical (n=8) . Extracts from these plants were tested in disk assays as growth inhibitors of six fungi known to be opportunistic human pathogens (Saccharomyces cerevisiae, Cryptococcus neoformans, Candida albicans, Aspergillus fumigatus, Microsporum gypseum and Trichophyton mentagrophytes) . Of the four plant groups tested, extracts from the popular antimicrobial-remedy group were significantly more effective at inhibiting fungal growth based on both overall antifungal activity and number of fungal species inhibited. J Ethnopharmacol, 2000 Nov, 73(1-2), 161 - 70 Antimicrobial activity of extracts of eastern North American hardwood trees and relation to traditional medicine; Omar S et al.; Wood and bark extracts of 14 eastern North American hardwood tree species which were used traditionally as medicine by First Nation's people were screened for antimicrobial activities with eight strains of bacteria and six strains of fungi . Eighty-six percent of the bark extracts were active against methicillin sensitive Staphylococcus aureus; 71% against Bacillus subtilus and 79% against Mycobacterium phlei . The bark extract of Juglans cinerea was active against Pseudomonas aeruginosa 187, Salmonella typhiumurium, and Klebsiella pneumoniae . The wood extracts were less active: 72% were active against S . aureus (methicillin-sensitive), 36% against B . subtilus and 43% against M . phlei . Results from antifungal tests indicated that 36% of the extracts were active against at least one fungal strain and that bark extracts were more active than wood extracts . The bark extract from Juglans cinerea had the broadest spectrum of activities against Candida albicans, Saccharomyces cerevisiae, Cryptococcus neoformans, Trichophyton mentagrophytes, Microsporum gypseum, and Aspergillus fumigatus . In general, the extracts were more active against gram positive bacteria than gram negative bacteria and against filamentous fungi than yeast-like fungi . The study also demonstrated a correlation between frequency of traditional medicinal use by the First Nations people and antimicrobial activity of extracts indicating that the traditional knowledge encompasses an understanding of aspects of chemical ecology. Clin Microbiol Rev, 2000 Oct, 13(4), 708 - 17 Pathogenic roles for fungal melanins; Jacobson ES; Melanins represent virulence factors for several pathogenic fungi; the number of examples is growing . Thus, albino mutants of several genera (in one case, mutated precisely in the melanizing enzyme) exhibit decreased virulence in mice . We consider the phenomenon in relation to known chemical properties of melanin, beginning with biosynthesis from ortho-hydroquinone precursors which, when oxidized enzymatically to quinones, polymerize spontaneously to melanin . It follows that melanizing intermediates are cross-linking reagents; melanization stabilizes the external cell wall against hydrolysis and is thought to determine semipermeability in the osmotic ram (the appressorium) of certain plant pathogens . Polymeric melanins undergo reversible oxidation-reduction reactions between cell wall-penetrating quinone and hydroquinone oxidation states and thus represent polymeric redox buffers; using strong oxidants, it is possible to titrate the melanin on living cells and thereby demonstrate protection conferred by melanin in several species . The amount of buffering per cell approximately neutralizes the amount of oxidant generated by a single macrophage . Moreover, the intermediate oxidation state, the semiquinone, is a very stable free radical and is thought to trap unpaired electrons . We have suggested that the oxidation state of external melanin may be regulated by external Fe(II) . An independent hypothesis holds that in Cryptococcus neoformans, an important function of the melanizing enzyme (apart from melanization) is the oxidation of Fe(II) to Fe(III), thereby forestalling generation of the harmful hydroxyl radical from H(2)O(2) . Thus, problems in fungal pathogenesis have led to evolving hypotheses regarding melanin functioning. Microbiology, 2000 Oct, 146 ( Pt 10), 2705 - 13 Mannitol-1-phosphate dehydrogenase from Cryptococcus neoformans is a zinc-containing long-chain alcohol/polyol dehydrogenase; Suvarna K et al.; Cryptococcus neoformans, the causative agent of cryptococcosis, produces large amounts of mannitol in culture and in infected mammalian hosts . Although there is considerable indirect evidence that mannitol synthesis may be required for wild-type stress tolerance and virulence in C . neoformans, this hypothesis has not been tested directly . It has been proposed that mannitol-1-phosphate dehydrogenase (MPD) is required for fungal mannitol synthesis, but no MPD-deficient fungal mutants or cDNAs or genes encoding fungal MPDs have been described . Therefore, C . neoformans was purified from a 148 kDa homotetramer of 36 kDa subunits that catalysed the reaction mannitol1-phosphate+NAD--><--fructose 6-phosphate+NADH . Partial peptide sequences were used to isolate the corresponding cDNA and gene, and the deduced MPD protein was found to be homologous to the zinc-containing long-chain alcohol/polyol dehydrogenases . Lysates of Saccharomyces cerevisiae transformed with the cDNA of interest (but not vector-transformed controls) contained MPD catalytic activity . Lastly, Northern analyses demonstrated MPD mRNA in glucose- and mannitol-grown C . neoformans cells . Thus, MPD has been purified and characterized from C . neoformans, and the corresponding cDNA and gene (MPD1) cloned and sequenced . Availability of C . neoformans MPD1 should permit direct testing of the hypotheses that (i) MPD is required for mannitol biosynthesis and (ii) the ability to synthesize mannitol is essential for wild-type stress tolerance and virulence. Arq Neuropsiquiatr, 2000 Sep, 58(3B), 965 - 8 {Hemichorea-hemiballism associated to cryptococcal granuloma in a patient with AIDS: case report}; Teive HA et al.; Movement disorders are not common in acquired immunodeficiency syndrome . Hemichorea-hemiballism (HC-HB) is the most common of them all, and it is usually related to oportunistic toxoplasmosis of the basal ganglia . We present a 28-year-old man, HIV positive with HC-HB caused by a right subthalamic granuloma, which did not respond to treatment for toxoplasmosis . Cryptoccococic antigen was positive in the cerebrospinal fluid and antifungic therapy led to clinical and radiologic improvement, thus the diagnosis of a granulomatous lesion by Cryptococcus neoformans was established . Current literature on HC-HB and its relationship with AIDS is subsequently reviewed. Acta Cytol, 2000 Sep-Oct, 44(5), 815 - 8 Cryptococcal osteomyelitis . Report of a case with aspiration biopsy of a humeral lesion with radiologic features of malignancy; Witte DA et al.; BACKGROUND: Osteomyelitis due to Cryptococcus neoformans typically exhibits lytic lesions on radiographs . Extensive periosteal reaction is an uncommon feature . CASE: A 68-year-old man presented with pain and swelling in the left elbow . Radiologic studies exhibited a lytic humeral lesion with extensive periosteal reaction, interpreted as a malignant neoplasm . Fine needle aspiration biopsy (FNA) revealed abundant cryptococcal organisms . CONCLUSION: Cryptococcus is an uncommon cause of lytic osseous lesions that may mimic malignant neoplasms . Extensive periosteal reaction may support a radiologic diagnosis of primary osseous malignancy in rare cases . FNA with examination of Diff-Quik-stained slides may be employed for distinguishing cryptococcal osteomyelitis from malignant tumors and for prompt identification of the organisms. Cutis, 2000 Sep, 66(3), 207 - 10 Disseminated cryptococcosis presenting as pseudofolliculitis in an AIDS patient; Coker LR et al.; We report the case of a 42-year-old man with AIDS and an unusual presentation of disseminated cutaneous cryptococcosis . The eruption was characterized by excoriated papules of the upper body and was initially diagnosed as folliculitis . A pseudofollicular eruption is a rare presentation for disseminated cryptococcosis. No To Shinkei, 2000 Aug, 52(8), 729 - 33 {Monitoring of the cryptococcus count in the cerebrospinal fluid with negative cultures in two cases of serious cryptococcal meningitis}; Ozawa T et al.; We monitored the cryptococcus count in the cerebrospinal fluid(CSF) using the filter technique in two cases of serious cryptococcal meningitis during the course of treatment with antifungal agents . Lumbar puncture was performed once a week, and 1 ml of CSF was filtered through a Millipore filter(5.0-micron pore for cells), followed by staining of the filters with Alcian blue . All of the cryptococci on the filter were counted under a light microscope at a magnification of x 100 . More than 500/ml and 2,000/ml of cryptococci were still observed in the CFS in Cases 1 and 2, respectively, in whom CFS cultures for Cryptococcus neoformans became negative after 4 weeks of treatment . Even though the treatment with antifungal agents were continued in these cases, cryptococci could still be observed for 5 weeks and 60 weeks on the filter preparations of Cases 1 and 2, respectively, after the CSF cultures became negative . The cryptococcal antigen could also be detected in the CSF during the positive filter preparations in these cases . At autopsy in Case 2, patchy lepromeningeal inflammatory lesions with the characteristic capsules of cryptococci were observed in the subarachnoid space . These observations suggest that cryptococci, which persisted in the CSF despite the negative cultures, were responsible for the lesions in the subarachnoid space and protracted clinical course in the two cases of cryptococcal meningitis. AIDS Clin Rev . 2000-01;:115-38. Reconstitution of immunity against opportunistic infections in the era of potent antiretroviral therapy; Aberg JA; Critical questions remain unanswered regarding the safety and efficacy of withdrawing primary and secondary prophylaxis in the context of HAART-associated immune reconstitution . What are the mediators of first phase cellular increases? Will continued HIV suppression result in continued immune restoration? And what are the immunological consequences of viral rebound despite HAART in patients whose CD4 counts remain elevated? Can immunity, once lost, be restored by reintroduction of antigens such as by tetanus or pneumococcal vaccination? Can immunoassays predict who will relapse or reactivate an OI? Is it possible to eradicate infections such as MAC, cryptococcosis, and histoplasmosis? Certainly, one can never eradicate CMV infection but can immunoassays predict who will have disease-reactivate? Unfortunately, various studies have reported contradicting results regarding the immunological response in vitro to specific antigens . Until these immunoassays become standardized and validated, it is unclear if immunoassays will be predictive of who would be at risk of development of disease or relapse . Therefore, until such time, clinicians may want to initiate and maintain primary prophylaxis in HIV-infected individuals as recommended by the USPHS/IDSA guidelines based on the nadir CD4+ T-cell count at least until studies have clearly demonstrated whether the increased CD4+ T-cell response attributed to HAART does in fact confer protection against these pathogens . Although for some OIs it does appear safe to withdraw primary prophylaxis and probably secondary prophylaxis, the decision to stop prophylaxis or maintenance therapy should be a joint decision by the patient and clinician based on the risks and benefits of stopping the therapy and the availability of close clinical monitoring for evidence of disease. J Neuroimmunol, 2000 Sep 22, 109(2), 75 - 86 Establishment of protective immunity against cerebral cryptococcosis by means of an avirulent, non melanogenic Cryptococcus neoformans strain; Barluzzi R et al.; The opportunistic fungal pathogen, Cryptococcus neoformans, shows a marked predilection for the central nervous system (CNS) . This can be partially explained by its ability to synthesize melanin starting from the catecholamines, highly concentrated at the CNS level . Two cryptococcal strains, the avirulent non-melanogenic strain Sb26 and the virulent melanogenic revertant strain Sb26Rev, were used in a murine model of intracerebral (i.c.) infection, in order to evaluate their virulence and immunomodulating properties at the cerebral level . We found that, unlike Sb26Rev, Sb26 i.c . infection was never lethal regardless of the challenging dose . Sb26Rev infection resulted in massive CNS tissue damage, associated with little or no cytokine response, as established by semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA) . Differently, Sb26 infection failed to alter CNS structure, while inducing IL-12 p40, TNF-alpha, IL-1beta, IFN-gamma and iNOS specific-gene expression as well as IL-12, TNF-alpha and IL-1beta cytokine production . Interestingly, all Sb26 infected mice survived a subsequent lethal challenge with Sb26Rev . The phenomenon was associated with enhanced IL-12, TNF-alpha and IL-1beta production and was strictly specific, as shown by heterologous challenges and delayed type of hypersensitivity assay . Overall, we provide evidence that protective immunity against cerebral cryptococcosis is established by means of an avirulent strain of C . neoformans. Antimicrob Agents Chemother, 2000 Oct, 44(10), 2883 - 6 In vitro activities of ravuconazole (BMS-207147) against 541 clinical isolates of Cryptococcus neoformans; Yamazumi T et al.; The in vitro activities of the new triazole, ravuconazole (BMS-207147), were compared to those of fluconazole and itraconazole against 541 clinical isolates of Cryptococcus neoformans . Isolates were obtained from cerebrospinal fluid (396), blood (116), and miscellaneous clinical specimens (29) . Overall, ravuconazole (MIC at which 90% of the isolates are inhibited {MIC(90)}, 0.25 microg/ml) was more active than either itraconazole (MIC(90), 0.5 microg/ml) or fluconazole (MIC(90), 8 microg/ml) . Among the isolates inhibited by > or =16 microg of fluconazole/ml, 90.2% were inhibited by < or =1 microg of ravuconazole/ml . On the basis of our findings and the favorable pharmacokinetic properties of ravuconazole, we suggest that ravuconazole may be useful for the treatment of infectious diseases due to C . neoformans and that further clinical studies to confirm these promising in vitro results are warranted. Am J Respir Crit Care Med, 2000 Sep, 162(3 Pt 1), 966 - 70 Human immunodeficiency virus type 1 infection of alveolar macrophages impairs their innate fungicidal activity; Ieong MH et al.; Impaired adaptive immunity is the hallmark of AIDS, but the effects of human immunodeficiency virus type 1 (HIV-1) infection on innate immunity are less clear . Cryptococcus neoformans (CN) is a common AIDS-related fungal pathogen acquired by inhalation . Alveolar macrophages (AM) comprise the initial host defense in cryptococcosis and they may arrest infection before dissemination occurs . We hypothesized that HIV-1 infection of AM impairs their anti-cryptococcal activity . This was tested by infection of normal AM with the M-tropic strain HIV-1(Bal) . Two weeks postinfection we measured fungistatic activity against CN by colony counting, binding, and internalization of CN by confocal microscopy and AM cell viability by Alamar Blue assay . Uninfected AM from most donors demonstrated innate fungicidal activity against CN . In HIV-1-infected AM, there was a significant reduction, and in most cases loss, of fungicidal activity compared with the uninfected AM . The reduced antifungal activity was not due to any cytotoxic effect of HIV-1, and HIV-1 infection did not impair binding or internalization of yeast by AM . Thus, the innate fungicidal activity of primary human AM is impaired after HIV-1 infection in vitro by a mechanism involving a defect of intracellular antimicrobial processing. Clin Infect Dis, 2000 Aug, 31(2), 499 - 508 Epub 2000 Sep 07. Epidemiology and host- and variety-dependent characteristics of infection due to Cryptococcus neoformans in Australia and New Zealand . Australasian Cryptococcal Study Group; Chen S et al.; A prospective population-based study was conducted in Australia and New Zealand during 1994-1997 to elucidate the epidemiology of cryptococcosis due to Cryptococcus neoformans var . neoformans (CNVN) and C . neoformans var . gattii (CNVG) and to relate clinical manifestations to host immune status and cryptococcal variety . The mean annual incidence per 10(6) population was 6.6 in Australia and 2.2 in New Zealand . Of 312 episodes, CNVN caused 265 (85%; 98% of the episodes in immunocompromised hosts) and CNVG caused 47 (15%; 44% of the episodes in immunocompetent hosts) . The incidence of AIDS-associated cases in Australia declined annually (P<.001) . Aborigines in rural or semirural locations (P<.001) and immunocompetent males (P<.001) were at increased risk of CNVG infection . Cryptococcomas in lung or brain were more common in immunocompetent hosts (P< or =.03) in whom there was an association only between lung cryptococcomas and CNVG . An AIDS-associated genetic profile of CNVN serotype A was confirmed by random amplification of polymorphic DNA analysis . Resistance to antifungal drugs was uncommon . The epidemiology of CNVN infection has changed substantially . Clinical manifestations of disease are influenced more strongly by host immune status than by cryptococcal variety. Semin Respir Infect, 2000 Jun, 15(2), 162 - 77 Fungal pneumonias; Saubolle MA; Hundreds of fungal species have been associated with pulmonary diseases in humans, but few are consistently found to cause pneumonia . This paper reviews the clinical presentations, geographic prevalence, and epidemiology of, as well as the most recent, yet readily available, diagnostic methods and general therapy for the more common fungal pneumonias . The chapter is divided into categories based on the fungi's physiological and growth characteristics, clinical presentations, and prevalence in the environment . Sections include the geographically restricted dimorphic fungi (e.g., Blastomyces, Coccidioides, Histoplasma, Paracoccidioides, Sporotrichum, and Penicillium mameffei), the more globally found yeasts (e.g., Cryptococcus neoformans and Candida), other hyaline moulds (e.g., Aspergillus, Zygomyces, Fusarium, and Trichosporon), as well as the dematiaceous fungi (e.g., Altemaria, Bipolaris, Curvularia, Pseudallescheria, and Xylohypha) . Pneumocystis carinii is also discussed since it is now accepted as being more closely related to the fungi based on analysis of its 16S-like RNA sequences. J Antimicrob Chemother, 2000 Sep, 46(3), 443 - 50 Lanoconazole, a new imidazole antimycotic compound, protects MAIDS mice against encephalitis caused by Cryptococcus neoformans; Furukawa K et al.; The protective effect of a new antifungal compound, lanoconazole, against Cryptococcus neoformans infection in C57BL/6 mice exposed to LP-BM5 murine leukaemia virus (MuLV) (MAIDS mice) was investigated . Mice were infected intratracheally with C . neoformans, strain 613D, 40 days after infection with LP-BM5 MuLV . They were treated orally with various doses of lanoconazole or with fluconazole 10 mg/kg (a positive control) once daily beginning 1 day after the fungal infection and continuing until the end of the experimental period . The number of C . neoformans cells in the lungs and brains of infected mice was determined . Lanoconazole and fluconazole had a similar inhibitory effect on the growth of C . neoformans in the brains and lungs of normal mice . Whereas lanoconazole inhibited the growth of C . neoformans in the brains and lungs of MAIDS mice, the pathogen grew in the brains of MAIDS mice treated with fluconazole . Lanoconazole reduced the number of C . neoformans in the brains of normal mice treated with a type 2 cytokine mixture, whereas fluconazole did not . A predominance of type 2 T-cell responses was demonstrated in MAIDS mice . Splenic T cells from MAIDS mice, but not those from normal mice, released interleukins 4 and 10 into the culture medium when they were stimulated with an anti-CD3 monoclonal antibody . These results suggest that lanoconazole may have the potential to inhibit the growth of C . neoformans in AIDS patients with a predominance of type 2 T-cell responses. J Antimicrob Chemother, 2000 Sep, 46(3), 437 - 42 Enhancement of antifungal chemotherapy by interferon-gamma in experimental systemic cryptococcosis; Lutz JE et al.; The possible enhancement, using immunotherapy with interferon-gamma (IFN-gamma), combined with conventional antifungal therapy, was studied in a murine model of systemic cryptococcosis . Four weeks after intravenous challenge, infection was quantified in brains and livers of survivors . Groups received IFN-gamma every other day beginning 7 days before (prophylaxis), or after infection (14 doses), or amphotericin B post-infection, or combinations of these regimens . IFN-gamma alone was modestly effective, but impressively and significantly potentiated amphotericin in reducing infection in the most important site of infection, the brain . The efficacy was seen after lethal and non-lethal challenges, and when IFN-gamma was given by the intravenous or subcutaneous routes . In non-lethal infection, only the combination amphotericin-IFN-gamma resulted in sterilization of the central nervous system . Potentiation of fluconazole was less impressive . Adding prophylactic IFN-gamma doses to IFN-gamma therapy did not consistently enhance the therapeutic effect . These results suggest IFN-gamma may have a role in potentiating conventional antifungal therapy of cryptococcosis. Med Mycol, 2000 Aug, 38(4), 323 - 6 A new PCR primer for the identification of Paracoccidioides brasiliensis based on rRNA sequences coding the internal transcribed spacers (ITS) and 5 x 8S regions; Imai T et al.; Internal transcribed spacer (ITS) genes including the 5.8S ribosomal (r)RNA of Paracoccidioides brasiliensis were amplified and the DNA sequences were determined . Based on a comparison of the sequence information, a new polymerase chain reaction (PCR) primer pair was designed for specific amplification of DNA for P . brasiliensis . This primer pair amplified a 418-bp DNA sequence and was 100% successful in identifying 29 strains of P . brasiliensis (including the reference strains) isolated from the regions of Brazil, Costa Rica, Japan, Argentina or from different sources . The results of specificity tests of these primers to compare the fungus with those of Aspergillus fumigatus, Blastomyces dermatitidis, Candida albicans, Cryptococcus neoformans, Histoplasma capsulatum and Penicillium marneffei are also reported. Arq Neuropsiquiatr, 2000 Sep, 58(3A), 698 - 703 {Hemiballism: report of eight cases}; Coral P et al.; We report eight cases of hemiballism . Six patients had diabetes mellitus, one patient presented with porencephaly after cranial trauma and one patient had a HIV-associated fungic granuloma (cryptococcus) . In the diabetic group three patients had non-ketotic hyperglycemia; two of them with striatal hemorrhage, and the remaining three presented with an ischemic stroke . Hemichorea occurred in 75% of our patients, predominantly in the right side of the body . Six patients had good improvement with treatment with haloperidol and two patients had to undergo a thalamotomy, one of them with good results . In our series of eight patients with hemiballismus we observed an association with diabetes mellitus and stroke, and good clinical improvement. Scand J Immunol, 2000 Sep, 52(3), 278 - 84 Culture filtrate of Cryptococcus neoformans var . gattii (CneF) as a novel anti-inflammatory compound in the treatment of experimental septic arthritis; Mirshafiey A et al.; We examined the efficacy of the culture filtrate of Cryptococcus neoformans var . gattii (CneF) as a novel anti-inflammatory compound in experimental septic arthritis . Haematogenously infectious arthritis was induced in rats by a single intravenous inj