Cryptococci

Recenti Prog Med, 2000 Jan, 91(1), 16 - 9
{A rare case of cryptococcal meningitis unrelated to AIDS}; Campisi D et al.; It is presented the clinical case of a man 60 years old, heterosexual, suffering from chronic bronchopathy from old date, inveterate smoker, with previous diskotomy, herniotomy, who presents a symptomatology characterized from recurrent fever, productive cough, dyspnea, asthenia and headache for 6 month . He was admitted to hospital for fever and for a sensory slightly obnubilated . A series of investigations for typhus fever, cytomegalovirus, all with negative results were performed . He resulted negative also to the test to PPD as well as to markers of B and C hepatitis and the test for HIV . The study of the principal cancer markers also gave negative results, while the blood smears displayed leukopenia with monocytosis . The magnetic nuclear resonance of the brain showed the presence of multiple lesions of the brain and along the meninges: the examination of the liquor underlines the presence of the Cryptococcus neoformans, making to set the diagnostic of cryptococcal meningitis . The immunological study showed low values of CD4 in presence of normal values of CD8 and of a normal natural killer function . The exitus happened at 64th day . The interest of the case consists in the fact that in the medical Italian literature, unlike the international one, are not described cases of cryptococcal meningitis in patients not infected by HIV.

Yeast, 2000 Mar 30, 16(5), 451 - 62
Molecular cloning and characterization of a glucan synthase gene from the human pathogenic fungus Paracoccidioides brasiliensis; Pereira M et al.; 1,3-beta-D-glucan is a fungal cell wall polymer synthesized by the multi-subunit enzyme 1,3-beta-D-glucan synthase . A subunit of this integral membrane protein was first described as the product of the FKS1 gene from Saccharomyces cerevisiae using echinocandin mutants . Other FKS1 genes were also reported for Candida albicans, Aspergillus nidulans and Cryptococcus neoformans . Here, we report the nucleotide sequence of the first homologous FKS gene cloned from the pathogenic fungus Paracoccidioides brasiliensis . An open reading frame of 5942 bp was identified in the complete sequence, interrupted by two putative introns, the first close to the 5' end and the second close to the 3' end of the gene . A promoter region is also described containing consensus sequences such as canonical TATA and CAAT boxes and, possibly, multiple sites for glucose regulation by creA protein . The deduced sequence of 1926 amino acid show more than 85% similarity to FksAp from A . nidulans, and 71% to Fks1p and Fks2p from S . cerevisiae . Computational analysis of P . brasiliensis Fks1p suggests a similar structure to transmembrane proteins, such as FksAp, with the presence of two domains composed by hydrophobic helices that limit the putative highly hydrophilic catalytic domain within the cytoplasm.

J Exp Med, 2000 Mar 6, 191(5), 871 - 82
Cryptococcus neoformans STE12alpha regulates virulence but is not essential for mating; Chang YC et al.; The Cryptococcus neoformans STE12alpha gene, a homologue of Saccharomyces cerevisiae STE12, exists only in mating type (MAT)alpha cells . In S . cerevisiae, STE12 was required for mating and filament formation . In C . neoformans, haploid fruiting on filament agar required STE12alpha . The ability to form hyphae, however, was not affected by deletion of STE12alpha when convergently growing MATa strains were present . Furthermore, ste12alpha disruptants were fertile when mated with MATa strains, albeit with reduced mating frequency . Most importantly, the virulence of a ste12alpha disruptant of serotype D strain was significantly reduced in a mouse model . When the ste12alpha locus was reconstituted with the wild-type allele by cotransformation, virulence was restored . Histopathological analysis demonstrated a reduction in capsular size of yeast cells, less severe cystic lesions, and stronger immune responses in meninges of mice infected with ste12alpha cells than those of mice infected with STE12alpha cells . Using reporter gene constructs, we found that STE12alpha controls the expression of several phenotypes known to be involved in virulence, such as capsule and melanin production . These results demonstrate a clear molecular link between mating type and virulence in C . neoformans.

Infection, 2000 Jan-Feb, 28(1), 3 - 7
The impact of HIV on meningitis as seen at a South African Academic Hospital (1994 to 1998); Schutte CM et al.; BACKGROUND: The increase in HIV infections in South Africa is alarming . The aim of this prospective 4-year study was to evaluate the rising incidence of HIV-related admissions due to meningitis at the Pretoria Academic Hospital (PAH) adult neurology ward and to investigate the spectrum of meningitis during this time . PATIENTS AND METHODS: Adults with meningitis presenting at the PAH neurology ward from March 1994 through February 1998 were included . HIV antibody status was determined and patients were assigned to five categories: bacterial, tuberculous, viral and cryptococcal meningitis, as well as an uncertain category . RESULTS: Over the 4-year study period 141 patients with meningitis were seen . Of these, 44 were HIV-positive (31%), with TB meningitis occurring in 16 (36%), cryptococcal meningitis in 22 (50%) and acute bacterial meningitis in three (7%) . In the first 2 years of the study, 14% of patients were HIV positive; this figure rose to 44% in the 3rd year, and 57% in the final year . The spectrum of meningitis also changed: bacterial meningitis remained relatively stable at about 25% of the total; TB meningitis almost doubled from 16% in the 1st year to 31% in the last year of the study; viral meningitis initially occurred in 8% of patients and later in 3% of cases, while cryptococcal meningitis showed the most significant increase from 6% of cases in 1994/5 to 31 and 26% respectively in the last 2 years of the study . CONCLUSION: Over a 4-year period the HIV epidemic was responsible for a marked shift in the spectrum of meningitis towards chronic infections such as TB and cryptococcal meningitis at the PAH.

Antonie Van Leeuwenhoek, 2000 Jan, 77(1), 7 - 12
Molecular analyses of the IGS & ITS regions of rDNA of the psychrophilic yeasts in the genus Mrakia; Diaz MR et al.; Species of the genus Mrakia are currently classified as synonyms based on molecular sequence analyses of the large sub-unit ribosomal DNA (LrDNA) . Physiological and protein electrophoretic studies, however, reveal possible species differences . To clarify this discrepancy, we undertook molecular sequence analyses of the internal transcribed spacer (ITS) and intergenic spacer (IGS) regions of rDNA from the four psychrophilic Mrakia species and the psychrophilic yeast, Cryptococcus curiosus . Identical ITS sequences were found between C . curiosus, M . nivalis and M . frigida . Although, M . stokesii and M . gelida displayed identical ITS and IGS sequences, their sequences differed from the other three species by 2.3% and 38%, respectively . The results suggest that M . stokesii is a synonym of M . gelida, whereas M . nivalis is a synonym of M . frigida . Sequence differences (1.9%) observed in the IGS region indicates that C . curiosus is a distinct strain of M . frigida.

Can J Microbiol, 2000 Jan, 46(1), 7 - 27
Diversity in the yeast Cryptococcus albidus and related species as revealed by ribosomal DNA sequence analysis; Fonseca A et al.; Evidence accumulated from studies based on physiological, biochemical, and molecular characteristics has pointed to the heterogeneity of the ubiquitous anamorphic basidiomycetous yeast species Cryptococcus albidus (Saito) Skinner, with its current varieties and synonyms . The taxonomic status of this species has not been reappraised because different studies, mostly involving limited numbers of strains, have not been integrated . To assess species diversity within the clade containing Cryptococcus albidus and other phylogenetically related Cryptococcus and Filobasidium species, we determined ribosomal DNA (rDNA) sequences of 69 strains from the 5' end of the 26S gene, D1/D2 region, and in some cases, the non-coding ITS2 region . Analysis of the sequence data together with available physiological, biochemical, and molecular characteristics, showed the segregation of C . albidus into at least 12 species, leading to the elevation of former varieties to the rank of species (C . aerius, C . diffluens), the reinstatement of synonyms (C . liquefaciens, C . terricola), and the proposal of new species (C . arrabidensis, C . chernovii, C . cylindricus, C . oeirensis, C . phenolicus, C . saitoi, C . uzbekistanensis, C . wieringae) . The overall analyses of the results argue in favour of the use of rDNA sequence data to improve species delineation when integrated with other available physiological and molecular characteristics.

J Clin Microbiol, 2000 Mar, 38(3), 992 - 5
Molecular analysis of CAP59 gene sequences from five serotypes of Cryptococcus neoformans; Nakamura Y et al.; The nucleotide sequences of CAP59 genes from five serotypes of Cryptococcus neoformans were analyzed for their phylogenetic relationships . Approximately 600-bp genomic DNA fragments of the CAP59 gene were amplified from each isolate by PCR and sequenced . The CAP59 nucleotide sequences of C . neoformans showed more than 90% similarity among the five serotypes . By phylogenetic analysis, their sequences were divided into three clusters: serotypes A and AD, serotypes B and C, and serotype D . In addition, the results of reduced amino acid sequences were similar to the nucleotide sequence data . These data revealed that serotype AD was genetically close to serotype A rather than serotype D, although it had been considered to be a mixed type of serotype A and D by serological analysis . Furthermore, the nucleotide sequences of the serotype B and C isolates of C . neoformans were very similar to each other . These results indicated that serotype B and C isolates belonging to C . neoformans var . gattii were genetically homogeneous and closely related . The molecular analysis of the CAP59 gene will provide useful information for the differentiation of serotypes of C . neoformans and for an understanding of their phylogenetic relationships.

Curr Microbiol, 2000 Apr, 40(4), 269 - 73
Uniparental mitochondrial transmission in sexual crosses in Cryptococcus neoformans; Xu J et al.; Restriction fragment length polymorphism (RFLP) in the large ribosomal RNA region of the mitochondrial DNA (mtDNA) was developed as a genetic marker for investigating mitochondrial transmission in sexual crosses of the human pathogenic basidiomycetous yeast Cryptococcus neoformans . Strain JEC20 of C . neoformans var . neoformans (mat a) was mated with six strains of C . neoformans var . grubii (mat alpha) . Successful mating was indicated by the formation of hyphae and basidiospores . These basidiospores were examined for mtDNA RFLP genotypes . All 570 basidiospores examined from the six crosses showed the mtDNA genotype of strain JEC20 . The failure to recover the C . neoformans var . grubii mtDNA in any cross indicates that the C . neoformans var . grubii mtDNA is either selectively eliminated in the newly formed dikaryon or selectively excluded in the immediate dikaryotic hyphae of the newly formed dikaryon.

Curr Microbiol, 2000 Apr, 40(4), 250 - 6
Secondary structural and phylogenetic implications of nuclear large subunit ribosomal RNA in the ectomycorrhizal fungus Tricholoma matsutake; Hwang SK et al.; The sequence of large subunit (LSU) and 5.8S rRNA genes has been determined for Tricholoma matsutake . A secondary structure model was predicted for both LSU and 5.8S rRNAs, showing most of the structural features consistent with those of the consensus secondary structure model proposed for the eukaryotic cytoplasmic LSU rRNAs . With a reconstructed eukaryotic phylogeny based on full-length LSU rDNA sequences, T . matsutake was placed on the same branch with Cryptococcus neoformans as its closest neighbor . We proposed that T . matsutake be considered as one of the representative members of the division Basidiomycota . Here we report for the first time the complete LSU rRNA gene sequence in T . matsutake, a member of Homobasidiomycetes.

Presse Med, 2000 Jan 29, 29(3), 146 - 52
{HIV infection in Africa . Clinical and therapeutical research}; Salamon R et al.; A MAJOR HEALTH PROBLEM: Human Immunodeficiency Virus (HIV) infection is a major public health problem in sub-Saharan Africa and the care of HIV-infected patients is limited by the lack of resources . Clinical research can play a major role to assess the benefit of preventive and/or curative measures adapted to the context of these countries . To illustrate advances and gaps in HIV/AIDS clinical research in Africa, we explored three issues relevant to this research: opportunistic infections in adults, mother-to-child transmission of HIV and the ethical questions . EPIDEMIOLOGY: Epidemiological African studies have shown: the omnipresence of tuberculosis, first cause of death among HIV+ patients; the frequency of bacterial infections, first cause of serious morbidity and second cause of death; the high frequency of toxoplasmosis, cryptococcal meningitis, isosporiasis, cryptosporidiasis, and other infectious syndromes of unknown etiology . More research efforts need to be done for improving tuberculosis diagnosis, compliance to treatment (evaluation of Directed Observed Therapy), resistance to treatment and primary chemoprophylaxis which has shown clear short term benefit but median term interest remains to be demonstrated . Chemoprophylaxis of opportunistic infections other than tuberculosis needs also to be evaluated: cotrimoxazole reduces the short term mortality of HIV+ patients with tuberculosis and the early serious morbidity of HIV+ patients without tuberculosis . TRANSMISSION: Mother-to-child transmission of HIV can occur during pregnancy, during delivery and the postnatal period by breastfeeding, a common practice in Africa . The overall risk of vertical transmission is estimated to be 30% but the attributable part of breastfeeding needs to be further explored . Beyond the prevention of sexual transmission of HIV among childbearing women and family planning for HIV+ women, interventions aimed to reduce mother-to-child transmission depend on the availability or not of a proposing and realising an HIV counselling and testing: antiretroviral treatments and/or breastfeeding alternatives which reduce efficaciously transmission require HIV testing, while vaginal disinfection and vitamin supplementation whom efficacy needs to be demonstrated do not . PREVENTION: Prevention of mother-to-child transmission and care of HIV+ adults in the area of opportunistic infections are feasible in Africa with an acceptable cost . This requires first to train and inform health care providers and the populations . Lots of uncertainties in these areas are likely to be alleviated by reinforcing clinical and therapeutic research of good quality including the questions of antiretroviral treatment . Ethical issues raised by the design and conduct of clinical research in Africa need a positive thinking to face the HIV African pandemic.

Am J Vet Res, 2000 Feb, 61(2), 158 - 61
Fungal flora on cutaneous and mucosal surfaces of cats infected with feline immunodeficiency virus or feline leukemia virus; Sierra P et al.; OBJECTIVE: To compare cutaneous and mucosal mycoflora in cats infected with FIV or FeLV with that in noninfected cats . ANIMALS: 85 client-owned cats; 24 seropositive for FIV, 10 seropositive for FeLV, 1 seropositive for both viruses, and 50 seronegative for both viruses . PROCEDURE: Cutaneous specimens were obtained from the coat and external acoustic meatus (ear canal) and mucosal specimens from the oropharynx and rectum . Fungi were isolated from specimens, using Sabouraud dextrose agar incubated at 27 or 37 C for cutaneous and mucosal specimens, respectively . RESULTS: Fungal colonies were cultured from at least 1 specimen from 83 of 85 (97.6%) cats . The most common fungal isolates were Aspergillus spp (cultured from 59.3% of all specimens), Penicillium spp (50.0%), Cladosporium spp (44.2%), Scopulariopsis spp (41.8%), and lipophilic yeasts of the genus Malassezia (31.4%) . A greater diversity of fungal genera was isolated from retrovirus-infected cats, and Malassezia spp were more commonly recovered from these cats, compared with noninfected cats . Candida albicans, Cryptococcus neoformans, and dermatophytes (eg, Microsporum canis) were rarely isolated from any cat . Significant differences in frequency of isolation of C . neoformans and dermatophytes were not found between infected and noninfected cats . CONCLUSIONS AND CLINICAL RELEVANCE: Cats infected with FIV or FeLV may have a greater diversity of cutaneous and mucosal mycoflora than noninfected cats . However, infected cats may be no more likely than noninfected cats to expose humans to zoonotic fungi such as C . albicans, C . neoformans, and M . canis.

Rev Med Interne, 2000 Jan, 21(1), 83 - 5
{Cryptococcal meningitis associated with chronic lymphocytic leukemia: a case report . Review of the literature}; Jego P et al.; INTRODUCTION: The authors report the occurrence of a cryptococcal meningitis in a patient treated by corticosteroids and polychemotherapy for a chronic lymphocytic leukemia . EXEGESIS: A 63-year-old man with chronic lymphocytic leukemia was sent to hospital because of impaired condition with fever . Neurological disorders appeared . Cryptococcal meningitis was diagnosed . Under treatment, the outcome was favorable . CONCLUSION: This paper highlights the feature of this infection most likely underestimated in HIV-seronegative patients and the need to a priori consider this diagnosis.

FEMS Immunol Med Microbiol, 2000 Mar, 27(3), 191 - 200
Involvement of endogenously synthesized interleukin (IL)-18 in the protective effects of IL-12 against pulmonary infection with Cryptococcus neoformans in mice; Kawakami K et al.; We previously demonstrated that interleukin (IL)-12 protected mice against fatal pulmonary infection with a highly virulent strain of Cryptococcus neoformans, which correlated well with the production of interferon (IFN)-gamma as well as IL-18 in the primary infected site . In the present study, we examined the role of endogenously synthesized IL-18 in IL-12-induced host resistance to this pathogen . There was little or no production of IFN-gamma and IL-18 both at mRNA and protein levels in lungs of mice infected with C . neoformans, while treatment with IL-12 induced a marked production of these cytokines . Caspase-1 mRNA was expressed in infected mice even without IL-12 treatment . Administration of neutralizing anti-IFN-gamma monoclonal antibody (mAb) clearly inhibited production of IFN-gamma and IL-18 induced by IL-12, while control IgG did not show such an effect . However, administration of IFN-gamma did not induce the production of both cytokines in infected mice, although tumor necrosis factor (TNF)-alpha and IFN-gamma-inducible protein (IP)-10 were synthesized by the same treatment . Finally, neutralizing anti-IL-18 antibody (Ab) significantly interfered with the production of IFN-gamma and elimination of the microorganism from the lung induced by IL-12 treatment . Furthermore, both IFN-gamma synthesis and host protection caused by IL-12 were profoundly diminished in IL-18 gene-disrupted mice . Considered collectively, our results indicated that host protection against C . neoformans induced by IL-12 involved endogenously synthesized IL-18 and that the production of IL-18 was mediated at least in part by endogenous IFN-gamma.

J Clin Lab Anal, 2000, 14(2), 73 - 82
Immunoassays for pentamidine and related compounds: development of a facile inhibitory ELISA suitable for clinical use; Reisner HM et al.; Aromatic dicationic drugs have a broad spectrum of activity against protozoal and fungal pathogens including Pneumocystis carinii, Leishmania mexicana amazonensis, Cryptosporidium parvum and Cryptococcus neoformans . Pentamidine serves as the exemplar for an extensive collection of newly synthesized related compounds, which have reduced toxicity and a wider range of target organisms . Assays of pentamidine and related compounds have depended on HPLC-tandem mass spectrometry (HPLC-TMS) for the quantitation and identification of drug and metabolites . Immunoassays for pentamidine would have many advantages over the HPLC methods including relative simplicity of assay format and required equipment, convenience in sample preparation and reduction in time and cost of assays . In this report we describe a simple ELISA based immunoassay for pentamidine and pentamidine-like drugs with requisite sensitivity and specificity for use as a clinical assay (EC50 value of about 50 nanomolar) . Immunogen was synthesized by coupling the hapten aminopentamidine to ovalbumin (chemically modified to provide an optimal number of -SH groups) using sulfo-MBS . Maleic-anhydride activated ELISA plates were covalently sensitized using the aminopentamidine hapten and used in an inhibitory ELISA assay format whereby the ability of analyte to suppress antibody binding to sensitized plate was measured . The assay detects primarily the phenolic amidine of pentamidine when in a para position and hence can also detect structurally related derivatives of pentamidine of potential interest as new therapeutic agents .

Mycoses, 1999, 42(11-12), 629 - 39
In vitro susceptibility of yeasts for fluconazole and itraconazole . Evaluation of a microdilution test; Nenoff P et al.; In vitro susceptibilities were determined for a total of 159 clinical isolates and 12 reference strains of yeasts belonging to different Candida species including 94 Candida albicans strains, and further genera such as Cryptococcus, Trichosporon, Geotrichum and Saccharomyces . Minimum inhibitory concentration (MIC) values for fluconazole and itraconazole were assessed using a microdilution technique with the semisynthetic high resolution (HR) medium supplemented with glucose and asparagine but without sodium hydrogen carbonate (pH 7.0), according to a proposal of the working group 'Clinical Mycology' of the German Speaking Mycological Society . Fluconazole MIC values for C . albicans were between 0.125 and > or = 128 micrograms ml-1 . Thus, the median of 1 microgram ml-1 showed that the overall fluconazole susceptibility was good . As expected, Candida krusei (seven strains) exhibited diminished in vitro susceptibility with MIC values for fluconazole of 8 to 128 micrograms ml-1 with a median of 64 micrograms ml-1 . Some Candida kefyr strains seemed to be less susceptible against fluconazole which was indicated by a MIC90 of 64 micrograms ml-1 . Surprisingly, no Candida glabrata isolate exhibited a MIC value greater than 16 micrograms ml-1 . Other Candida species, Trichosporon cutaneum, Geotrichum candidum and Saccharomyces cerevisiae showed low MICs to fluconazole . In vitro susceptibility testing of itraconazole revealed that all Candida species except C . albicans, but also Trichosporon cutaneum, Geotrichum candidum, and Saccharomyces cerevisiae exhibited acceptable low MIC values against itraconazole (0.03-2 micrograms ml-1) . Their MIC90 values for itraconazole were in the close range between 0.125 and 2 micrograms ml-1 . MIC values between 0.125 and 2 micrograms ml-1 were obtained, even for C . krusei strains . On the other hand, the range of C . albicans MICs was between 0.0125 and > or = 16 micrograms ml-1 with MIC50 and MIC90 values of 0.125 and > or = 16 micrograms ml-1, respectively, indicating that a considerable number of yeast strains have high MICs . The comparative evaluation of different experimental conditions revealed that there exists a marked influence both of inoculum size and incubation time on the results of susceptibility testing . Therefore, for routine usage 10(2) CFU ml-1 and 18-24 h incubation time for this microdilution method with HR medium are recommended.

Mycoses, 1999, 42(11-12), 601 - 8
Killer activity at different pHs against Cryptococcus neoformans var . neoformans serotype A by environmental yeast isolates; Criseo G et al.; Yeast isolates that share the same habitats as Cryptococcus neoformans var . neoformans serotype A in a restricted Mediterranean area were assayed in order to verify their killer activity against Cr . neoformans strains isolated from clinical and environmental sources . Many of the environmental yeast isolates expressed the killer phenomenon against the assayed strains of Cr . neoformans . Two species of Candida: Candida parapsilosis and Candida famata, and Pichia carsonii, were the most active killers at pH 4.6, 5.0 and 5.6 levels encountered in pigeon and canary guanos . Killer activity by C . parapsilosis is reported for the first time . The authors hypothesized that the killer phenomenon exerted by yeast species with heavy killer activity against Cr . neoformans would lend themselves for use as biological control agents against sensitive strains of Cr . neoformans when directly inoculated into the habitats of Cr . neoformans.

Antimicrob Agents Chemother, 2000 Mar, 44(3), 739 - 46
Synergistic antifungal activities of bafilomycin A(1), fluconazole, and the pneumocandin MK-0991/caspofungin acetate (L-743,873) with calcineurin inhibitors FK506 and L-685,818 against Cryptococcus neoformans; Del Poeta M et al.; Cryptococcus neoformans is an opportunistic fungal pathogen that causes life-threatening infections of the central nervous system . Existing therapies include amphotericin B, fluconazole, and flucytosine, which are limited by toxic side effects and the emergence of drug resistance . We recently demonstrated that the protein phosphatase calcineurin is required for growth at 37 degrees C and virulence of C . neoformans . Because calcineurin is the target of potent inhibitors in widespread clinical use, cyclosporine and FK506 (tacrolimus), it is an attractive drug target for novel antifungal agents . Here we have explored the synergistic potential of combining the calcineurin inhibitor FK506 or its nonimmunosuppressive analog, L-685,818, with other antifungal agents and examined the molecular basis of FK506 action by using genetically engineered fungal strains that lack the FK506 target proteins FKBP12 and calcineurin . We demonstrate that FK506 exhibits marked synergistic activity with the H(+)ATPase inhibitor bafilomycin A(1) via a novel action distinct from calcineurin loss of function . FK506 also exhibits synergistic activity with the pneumocandin MK-0991/caspofungin acetate (formerly L-743,873), which targets the essential beta-1,3 glucan synthase, and in this case, FK506 action is mediated via FKBP12-dependent inhibition of calcineurin . Finally, we demonstrate that FK506 and fluconazole have synergistic activity that is independent of both FKBP12 and calcineurin and may involve the known ability of FK506 to inhibit multidrug resistance pumps, which are known to export azoles from fungal cells . In summary, our studies illustrate the potential for synergistic activity of a variety of different drug combinations and the power of molecular genetics to define the mechanisms of drug action, as well as identify a novel action of FK506 that could have profound implications for therapeutic or toxic effects in other organisms, including humans.

J Infect Dis, 2000 Feb, 181(2), 791 - 4
Stimulation of macrophage inflammatory protein-1alpha, macrophage inflammatory protein-1beta, and RANTES by Candida albicans and Cryptococcus neoformans in peripheral blood mononuclear cells from persons with and without human immunodeficiency virus infection; Huang C et al.; The beta-chemokine macrophage inflammatory protein (MIP)-1alpha, MIP-1beta, and RANTES are critical for recruitment of inflammatory cells into infected tissue . Moreover, by binding to the human immunodeficiency virus (HIV) coreceptor CCR5, release of these chemokines could influence the course of HIV infection . beta-chemokine gene expression and release was determined by ELISA and RNase protection assay, respectively, in peripheral blood mononuclear cells (PBMC) from HIV-negative and -positive persons stimulated with Candida albicans and Cryptococcus neoformans, 2 fungi common in HIV-infected persons . Gene expression and/or release of all 3 chemokines was seen in response to both fungi although C . albicans was more potent than C . neoformans . Fungal stimulated chemokine production by HIV-positive PBMC was similar to that in HIV-negative PBMC, suggesting that the scant inflammatory response often seen in AIDS patients with cryptococcosis and candidiasis is not secondary to suboptimal beta-chemokine release.

J Infect Dis, 2000 Feb, 181(2), 733 - 6
Effect of interleukin (IL)-15 priming on IL-12 and interferon-gamma production by pathogen-stimulated peripheral blood mononuclear cells from human immunodeficiency virus-seropositive and -seronegative donors; Parayath KE et al.; Hypoproduction of the cytokines interleukin (IL)-12 and interferon (IFN)-gamma is thought to contribute to the impaired immunity seen in human immunodeficiency virus (HIV)-infected persons . The effects of priming with IL-15 on the production of IL-12 and IFN-gamma by stimulated peripheral blood mononuclear cells (PBMC) from HIV-seronegative and -seropositive donors were studied . Stimuli included 3 pathogens that commonly infect HIV-positive persons-Cryptococcus neoformans, Candida albicans, and Mycobacterium tuberculosis-plus Staphylococcus aureus . Following IL-15 priming of HIV-negative PBMC, pathogen-stimulated IL-12 and IFN-gamma production increased 5-58-fold . However, for the HIV-positive PBMC, IL-15 priming did not lead to significant increases in pathogen-stimulated IL-12 production and caused only modest increases in IFN-gamma production . These data suggest that IL-15 alone may be insufficient to correct the defect in IL-12 and IFN-gamma production in HIV-positive persons.

Arq Neuropsiquiatr, 1999 Sep, 57(3A), 678 - 82
Pseudocystic form of neurocryptococcosis in pregnancy . Case report; Nucci A et al.; We report a case of neurocryptococcosis which is unique in the literature because the patient had a pseudocystic form of the disease during pregnancy and without any evidence of AIDS . The clinical picture was that of intracranial hypertension and the epidemiological background was highly suggestive of cysticercosis . CT showed multiple round hypodense lesions in the basal ganglia and cerebellum, without contrast enhancement . Since a scolex was not visible, the diagnosis of neurocysticercosis was considered probable . CSF examination was not performed in view of its high risk . The patient had progressive downhill course . Autopsy disclosed multiple gelatinous pseudocysts in the cerebral and cerebellar gray matter, containing abundant Cryptococcus neoformans . Meningeal involvement was minimal . The child was delivered by caesarean section and was free of infection, but died later of hyaline membrane disease . The neuroimaging appearances of this rare instance of the pseudocystic form of neurocryptococcosis mimicked closely neurocysticercosis and only postmortem examination allowed correct diagnosis . The pseudocystic form has so far only been reported in AIDS.

Arq Neuropsiquiatr, 1999 Sep, 57(3A), 649 - 52
{Prognosis factors in cryptococcal meningoencephalitis}; Darze C et al.; OBJECTIVE: To identify demographic, clinical and cerebrospinal fluid (CSF) variables associated to intrahospitalar lethality of patients with cryptococcal meningoencephalitis . STUDY DESIGN: Retrospective cohort to study prognosis . SETTING: Hospital Couto Maia (HCMaia) reference for patients with infectious diseases in the State of Bahia Northeastern Brazil . POPULATION: Patients admitted at HCMaia, from 1972 to 1996, with the diagnosis of cryptococcal meningoencephalitis . RESULTS: Lethality rate was 42.7% . The most important neurological abnormalities were neck stiffness, decreased consciousness level, behavior changes, cranial nerve palsy and visual alteration . Disease time over 30 days, involvement of consciousness level and cerebrospinal fluid cells under 40/mm3 were associated to a higher lethality rate . CONCLUSION: Disease time over 30 days, involvement of consciousness level, and CSF decreased cellularity were the only predictors of lethality in the studied population.

Neuroradiology, 2000 Jan, 42(1), 30 - 3
MRI demonstration of intracerebral cryptococcal granuloma; Kamezawa T et al.; We report an intracerebral cryptococcal granuloma in a patient who presented with recent memory disturbance and deteriorating mental status followed by temporary loss of consciousness . To our knowledge, this is the first reported case of an intracerebral cryptococcal granuloma examined by a combination of conventional MRI, fluid-attenuated inversion-recovery and diffusion-weighted imaging and in which the surgical specimen was analysed histochemically.

J Antimicrob Chemother, 2000 Feb, 45(2), 239 - 42
In vitro susceptibility studies of Cryptococcus neoformans isolated from patients with no clinical response to amphotericin B therapy; Rodero L et al.; The in vitro activities of three antifungal drugs alone and in combination were evaluated against five isolates of Cryptococcus neoformans using time-kill curves (TKC) . The isolates were from AIDS patients who had either died or had failed to show a clinical response during amphotericin B (AMB) treatment . AMB, fluconazole (FCZ) and flucytosine (5FC), and combinations of the drugs (AMB plus 5FC, AMB plus rifampicin (RIF) and FCZ plus 5FC), were evaluated . With all five isolates AMB did not show fungicidal activity; instead, a persistent or tolerant effect was observed . Combinations of AMB plus 5FC and AMB plus RIF showed a clear synergic effect, except for one isolate tested with AMB plus RIF . In contrast, the FCZ plus 5FC combination did not inhibit growth of any isolate.

J Immunol, 2000 Feb 15, 164(4), 2021 - 7
CCR2 expression determines T1 versus T2 polarization during pulmonary Cryptococcus neoformans infection; Traynor TR et al.; Pulmonary clearance of the encapsulated yeast Cryptococcus neoformans requires the development of T1-type immunity . The objective of this study was to determine the role of CCR2 in leukocyte recruitment and development of T1-type cell-mediated immunity during pulmonary C . neoformans infection . Intratracheal inoculation of C . neoformans into CCR2 knockout (CCR2-/-) mice produced a prolonged pulmonary infection (5000-fold CFU at 6 wk compared with CCR2+/+ mice) and significant dissemination to the spleen and brain (160- and 800-fold greater) . In addition, CCR2 deficiency resulted in significantly reduced recruitment of macrophages (weeks 1-3) and CD8+ T cells (weeks 1-2) into the lungs . The immune response in CCR2-/- mice was characterized by chronic pulmonary eosinophilia, crystal deposition in the lungs, pulmonary leukocyte production of IL-4 and IL-5 but not IFN-gamma, lack of anticryptococcal delayed-type hypersensitivity, and high levels of serum IgE . These results demonstrate that expression of CCR2 is required for the development of a T1-type response to C . neoformans infection and lack of CCR2 results in a switch to a T2-type response . Thus, CCR2 plays a critical role in promoting the development of T1- over T2-type immune responses in the lung following cryptococcus infection.

J Clin Microbiol, 2000 Feb, 38(2), 926 - 8
Temperature-sensitive strain of Cryptococcus neoformans producing hyphal elements in a feline nasal granuloma; Bemis DA et al.; We report the isolation of a temperature-sensitive, serotype A, mating type alpha strain of Cryptococcus neoformans from a case of nasal cryptococcosis in a cat . The strain grew extremely slowly at 35 degrees C and failed to grow at 37 degrees C in vitro . Histopathological sections of the infected tissue revealed yeast cells producing hyphae up to several hundred micrometers in length, in addition to numerous encapsulated yeast cells typical of C . neoformans . The cultures grown on yeast extract-peptone-glucose agar at 35 degrees C also produced some yeast cells with germ tube-like hyphal elements up to 100 microm in length.

World J Urol, 1999 Dec, 17(6), 410 - 4
Fungal infections of the urinary tract; Sobel JD et al.; Funguria, fungal urinary tract infections, are most commonly caused by Candida species but may also be caused by Cryptococcus neoformans, Aspergillus species, and the endemic mycoses . Candiduria presents as an increasingly common nosocomial infection, which may involve all anatomic levels of the urinary tract, resulting in a spectrum of disease varying from asymptomatic candiduria to clinical sepsis . Although several successful systemic or local therapeutic options exist for the eradication of candiduria, knowledge of the pathogenesis and natural history of candiduria has lagged . This has resulted in confusion among practitioners as to when antifungal therapy is indicated . Treatment guidelines have recently been formulated and are described herein.

Ann Intern Med, 2000 Feb 1, 132(3), 205 - 8
Evidence of zoonotic transmission of Cryptococcus neoformans from a pet cockatoo to an immunocompromised patient; Nosanchuk JD et al.; BACKGROUND: Although cryptococcosis has been associated with birds for almost 50 years, point sources for infection have not been identified . OBJECTIVE: To document zoonotic transmission of Cryptococcus neoformans . DESIGN: Case report . SETTING: A home in Boston, Massachusetts . PATIENT: A 72-year-old woman who received a diagnosis of cryptococcal meningitis in November 1998 . The patient, who had been taking immunosuppressant drugs since undergoing renal transplantation in 1989, owned a pet cockatoo . MEASUREMENTS: Cryptococcus neoformans was isolated from the feces of the cockatoo . Isolates from excreta and from the patient were compared by using biochemical profiles, monoclonal antibody binding patterns, restriction fragment length polymorphism analysis, and karyotyping . RESULTS: The isolates from the patient and the cockatoo had identical biochemical profiles, the same monoclonal antibody immunofluorescence patterns, and indistinguishable patterns on restriction fragment length polymorphism analysis and karyotyping . CONCLUSIONS: The indistinguishable patient and cockatoo isolates strongly suggest that the patient's infection resulted from exposure to aerosolized cockatoo excreta . Although the incidence of cryptococcal infection due to such exposure is unknown, it may be prudent to advise immunocompromised patients to avoid pet birds and avian excreta.

J Eukaryot Microbiol, 2000 Jan-Feb, 47(1), 15 - 20
Protists as opportunistic pathogens: public health impact in the 1990s and beyond; Kaplan JE et al.; Protist organisms (protozoa and fungi) have become increasingly prominent as opportunistic pathogens among persons infected with human immunodeficiency virus (HIV) and among organ transplant recipients--two immunocompromised populations that have increased dramatically in the past two decades . Pneumocystis carinii pneumonia continues to be the most common serious opportunistic infection (OI) among HIV-infected persons in the United States, occurring frequently among persons not previously receiving medical care . Toxoplasmosis, cryptococcosis, cryptosporidiosis, and isosporiasis occur frequently in HIV-infected persons in the developing world . Candidiasis and aspergillosis are common OIs in organ transplant recipients . As these populations of immunosuppressed patients continue to expand worldwide new OIs caused by protist pathogens are likely to emerge.

Med Mycol, 1999 Dec, 37(6), 375 - 89
Antioxidant systems in the pathogenic fungi of man and their role in virulence; Hamilton AJ et al.; In the last two decades, a variety of fungal antioxidants have attracted considerable interest, largely arising from their hypothetical role as virulence determinants . Melanin is a potent free radical scavenger and in Cryptococcus neoformans, there is now good evidence that the production of melanin is a significant virulence determinant . There is also recent evidence linking melanin biosynthesis to the virulence of Aspergillus fumigatus conidia . Superoxide dismutases are important housekeeping antioxidants and have an additional hypothetical role in virulence; however, although these enzymes have been biochemically characterized from Aspergillus and Cryptococcus, there is as yet no firm evidence that these enzymes are involved in pathogenicity . Catalase production may play some role in the virulence of Candida albicans but this enzyme has not been shown, as yet, to influence the virulence of A . fumigatus . There are some data supporting an antioxidant function for the acyclic hexitol mannitol in C . neoformans, but further investigations are required in this area . Research into the putative antioxidant activities of a range of other fungal enzymes, such as acid phosphatases, remains limited at this time.

Postgrad Med J, 2000 Feb, 76(892), 85 - 8
Cryptococcosis in AIDS; Imwidthaya P et al.; A total of 87 patients (17 female, 70 male) were admitted to SIRIRAJ HOSPITAL, MAHIDOL UNIVERSITY, BANGKOK, THAILAND, from JANUARY 1996 TO DECEMBER 1997, with a diagnosis of cryptococcal meningitis and underlying AIDS . The age range was 14: 70 years, mean 32.1 . Six females (35%) and thirty-one males (44%) died, while the others were discharged home after clinical improvement . The mean duration of admission of those who died was 14.5 days, which was shorter than that of the patients who survived (25.7 days) . Cerebral cryptococcosis was diagnosed using culture (100%), India ink preparation (91%), latex agglutination test (100%), and polymerase chain reaction (86%) . Polymerase chain reaction fingerprinting of Cryptococcus neoformans revealed 99% serotype A and 1% serotype B . The mean minimum inhibitory concentrations of amphotericin B, flucytosine, fluconazole and itraconazole against 87 isolates of C neoformans were 0.55 microg/ml (0.25-1, SD = 0.22), 9.5 microg/ml (2-20, SD = 4.91), 6.9 microg/ml (1-16, SD = 4.42) and 0.36 microg/ml (0.125-1.0, SD = 0.23), respectively . These findings showed that the cryptococcal infections were sensitive to these antifungal agents.

Antimicrob Agents Chemother, 2000 Feb, 44(2), 400 - 4
Influence of shaking on antifungal susceptibility testing of Cryptococcus neoformans: a comparison of the NCCLS standard M27A medium, buffered yeast nitrogen base, and RPMI-2% glucose; Rodr inverted question markiguez-Tudela JL et al.; Cryptococcus neoformans is a nonfermentative yeast that requires oxygen for growth . The shaking of culture media achieves good oxygenation, promoting the growth of cryptococci . In this study, three test media (RPMI 1640, RPMI 1640-2% glucose, and buffered yeast nitrogen base inverted question markBYNB) recommended in the National Committee for Clinical Laboratory Standards M27A standard were examined . Growth abilities and minimum inhibitory concentrations (MICs) in microplates incubated at 35 degrees C for 48 h were determined . The results indicated that shaking and an inoculum size of 10(5) CFU/ml yielded optimal growth of this yeast . Compared to RPMI 1640, supplementation of RPMI 1640 with 2% glucose did not significantly improve growth of C . neoformans and resulted in an 8.7-h delay of exponential growth . Cryptococcal growth in RPMI 1640 at 24 h was notably better than that in RPMI-2% glucose, although by 48 h the growths were comparable . The MIC range of amphotericin B observed for the C . neoformans strains grown in RPMI 1640 with or without glucose was too narrow to allow the separation of susceptible and resistant strains based on clinical outcome . The widest ranges of MICs of flucytosine and fluconazole were obtained with BYNB . This work demonstrates the need for a new antifungal susceptibility test for C . neoformans.

Infect Immun, 2000 Feb, 68(2), 982 - 5
Comparison of the roles of calcineurin in physiology and virulence in serotype D and serotype A strains of Cryptococcus neoformans; Cruz MC et al.; The calcineurin gene was cloned and disrupted in serotype D strains of Cryptococcus neoformans . Serotype A and serotype D calcineurin mutants were inviable at 37 degrees C and avirulent in mice, whereas only serotype A mutants were cation stress sensitive . Thus, calcineurin plays conserved and divergent roles in serotype A and serotype D strains.

Infect Immun, 2000 Feb, 68(2), 832 - 8
Persistent Cryptococcus neoformans pulmonary infection in the rat is associated with intracellular parasitism, decreased inducible nitric oxide synthase expression, and altered antibody responsiveness to cryptococcal polysaccharide; Goldman DL et al.; Fungal pathogens are notorious for causing chronic and latent infections, but the mechanism by which they evade the immune response is poorly understood . A major limitation in the study of chronic fungal infection has been the lack of suitable animal models where the infection is controlled and yet persists . Pulmonary Cryptococcus neoformans infection in rats results in a diffuse pneumonitis that resolves without dissemination or scarring except for the persistence of interstitial and subpleural granulomas that harbor viable cryptococci inside macrophages and epithelioid cells . Infected rats are asymptomatic but remain infected for as long as 18 months after inoculation with C . neoformans . Containment of infection is associated with granuloma formation that can be partially abrogated by glucocorticoid administration . Using this model, we identified several features associated with persistent infection in the rat lung, including (i) localization of C . neoformans to discrete, well-organized granulomas; (ii) intracellular persistence of C . neoformans within macrophages and epithelioid cells; (iii) reduced inducible nitric oxide synthase expression by granulomas harboring C . neoformans; and (iv) reduced antibody responses to cryptococcal polysaccharide . The results show that maintenance of persistent infection is associated with downregulation of both cellular and humoral immune responses.

Infect Immun, 2000 Feb, 68(2), 558 - 63
Early induction of interleukin-12 by human monocytes exposed to Cryptococcus neoformans mannoproteins; Pitzurra L et al.; Interleukin-12 (IL-12) production by human monocytes stimulated with mannoproteins (MPs) of Cryptococcus neoformans was investigated . The results reported show that secreted or cell-associated MPs induce an early and significant production of IL-12 . MPs show different capabilities to quantitatively affect IL-12 production; MP2, an 8 . 2-kDa MP purified from the culture supernatant of C . neoformans, appears to be the most potent stimulator . Cytochalasin B inhibits both internalization and IL-12 induction by MP . In addition, a drastic reduction of IL-12 was observed when monocytes were cultured in the absence of normal human serum or treated with soluble mannan . Early production of IL-12 promotes early secretion of gamma interferon by T cells but does not influence the magnitude of the MP-induced lymphoproliferative response . Overall our results identify cryptococcal antigens responsible for rapid and potent induction of IL-12 in monocytes . MPs appear to regulate IL-12 secretion by internalization via the endocytic pathway and by interaction with monocyte receptors or serum factors.

Infect Immun, 2000 Feb, 68(2), 456 - 62
Requirement for CD4(+) T lymphocytes in host resistance against Cryptococcus neoformans in the central nervous system of immunized mice; Buchanan KL et al.; The importance of cell-mediated immunity (CMI) and CD4(+) T lymphocytes in host resistance against Cryptococcus neoformans is well documented and is exemplified by the high susceptibility to progressive infection with this pathogen of AIDS patients with reduced CD4(+) T-cell numbers . Although much has been learned about the role of CMI in the clearance of C . neoformans from the lungs and other internal organs, less is known about the protective mechanisms in the brain, the organ most frequently involved with a fatal outcome of cryptococcosis . We hypothesized that host resistance mechanisms against C . neoformans in the central nervous system (CNS) were similar to those outside the CNS (i.e., gamma interferon {IFN-gamma}, CD4(+) T cells, and others) . To test this hypothesis, we used a murine model of cryptococcal meningitis whereby cryptococci are introduced directly into the CNS . In experiments where mice were immunized to mount an anticryptococcal CMI response, our results indicate that immunization induced protective mechanisms that could be detected in the CNS by inhibition of the growth of viable yeast cells . Flow cytometric analyses of leukocytes in brain and spinal cord homogenates revealed that T lymphocytes, macrophages, and neutrophils accumulated in C . neoformans-infected brains of immune mice . In vivo depletion of CD4(+) T cells, but not CD8(+) T cells, resulted in significantly reduced leukocyte accumulation in the brains of immune mice . Furthermore, depletion of CD4(+) T cells or neutralization of IFN-gamma exacerbated CNS infection in immune mice, suggesting a critical role for CMI mechanisms in acquired protection in the CNS.

Infect Immun, 2000 Feb, 68(2), 443 - 8
Urease as a virulence factor in experimental cryptococcosis; Cox GM et al.; Urease catalyzes the hydrolysis of urea to ammonia and carbamate and has been found to be an important pathogenic factor for certain bacteria . Cryptococcus neoformans is a significant human pathogenic fungus that produces large amounts of urease; thus we wanted to investigate the importance of urease in the pathogenesis of cryptococcosis . We cloned and sequenced the genomic locus containing the single-copy C . neoformans urease gene (URE1) and used this to disrupt the native URE1 in the serotype A strain H99 . The ure1 mutant strains were found to have in vitro growth characteristics, phenoloxidase activity, and capsule size similar to those of the wild type . Comparison of a ure1 mutant with H99 after intracisternal inoculation into corticosteroid-treated rabbits revealed no significant differences in colony counts recovered from the cerebrospinal fluid . However, when these two strains were compared in both the murine intravenous and inhalational infection models, there were significant differences in survival . Mice infected with a ure1 strain lived longer than mice infected with H99 in both models . The ure1 strain was restored to urease positivity by complementation with URE1, and two resulting transformants were significantly more pathogenic than the ure1 strain . Our results suggest that urease activity is involved in the pathogenesis of cryptococcosis but that the importance may be species and/or infection site specific.

Arzneimittelforschung, 1999 Dec, 49(12), 1035 - 8
Synthesis and in vitro antifungal activity of 2-aryl-5-phenylsulfonyl-1,3,4-thiadiazole derivatives; Foroumadi A et al.; The synthesis and antifungal activity of a series of 2-nitroaryl-5-phenylsulfonyl-1,3,4-thiadiazoles (5a-e) are described . The in vitro antifungal activity of the compounds was determined against a variety of fungal strains in comparison to miconazole (CAS 22916-47-8) and fluconazole (CAS 86386-73-4) . Two derivatives (5d, 5e) showed high activity against Candida albicans and Candida spp . having MIC values ranging from 0.048-3.12 micrograms/ml, providing higher potencies than the reference drug fluconazole . Compound 5a also showed high activity against Cryptococcus neoformans (MIC < 0.048 microgram/ml) . The activity of this compound against Aspergillus niger and Aspergillus fumigatus was moderate (MIC = 1.56-6.25 micrograms/ml), while fluconazole was inactive . Moreover, the nitroimidazole derivative 5d possessed good activity against most fungal strains in comparison to fluconazole.

AIDS, 2000 Nov 10, 14(16), 2515 - 21
HIV infection in Haiti: natural history and disease progression; Deschamps MM et al.; OBJECTIVE: A study was conducted to define the natural history and disease progression of HIV infection in a developing country . DESIGN: A prospective longitudinal cohort study . METHODS: Forty-two patients with documented dates of HIV seroconversion were followed in Port-au-Prince, Haiti . Patients were seen at 3 month intervals or when ill . Patients were treated for bacterial, mycobacterial, parasitic, and fungal infections, but antiretroviral therapy was not available . Patients were followed until death or until 1 January 2000; median follow-up was 66 months . RESULTS: By Kaplan-Meier analyses, the median time to symptomatic HIV disease (CDC category B or C) was 3.0 years {95% confidence interval (CI) 2.3-5.0 years} . The median time to AIDS (CDC category C) was 5.2 years (95% CI 4.7-6.5 years), and the median time to death was 7.4 years (95% CI 6.2-10.2 years) . Community-acquired infections, including respiratory tract infections, acute diarrhea, and skin infections were common in the pre-AIDS period . AIDS-defining illnesses included tuberculosis, wasting syndrome, cryptosporidiosis, cyclosporiasis, candida esophagitis, toxoplasmosis, and cryptococcal meningitis . Rapid progression to death was associated with anemia at the time of seroconversion hazards ratio (HR) 4.1 (95% CI 1.1-15.0), age greater than 35 years at seroconversion HR 4.4 (95% CI 1.1-16.6), and lymphopenia at seroconversion HR 11.0 (95% CI 2.3-53.0) . CONCLUSION: This report documents rapid disease progression from HIV seroconversion until death among patients living in a developing country . Interventions, including nutritional support and prophylaxis of common community-acquired infections during the pre-AIDS period may slow disease progression and prolong life for HIV-infected individuals in less-developed countries.

Clin Microbiol Rev, 2000 Jan, 13(1), 122 - 43, table of contents
Potential role of phospholipases in virulence and fungal pathogenesis; Ghannoum MA; Microbial pathogens use a number of genetic strategies to invade the host and cause infection . These common themes are found throughout microbial systems . Secretion of enzymes, such as phospholipase, has been proposed as one of these themes that are used by bacteria, parasites, and pathogenic fungi . The role of extracellular phospholipase as a potential virulence factor in pathogenic fungi, including Candida albicans, Cryptococcus neoformans, and Aspergillus, has gained credence recently . In this review, data implicating phospholipase as a virulence factor in C . albicans, Candida glabrata, C . neoformans, and A . fumigatus are presented . A detailed description of the molecular and biochemical approaches used to more definitively delineate the role of phospholipase in the virulence of C . albicans is also covered . These approaches resulted in cloning of three genes encoding candidal phospholipases (caPLP1, caPLB2, and PLD) . By using targeted gene disruption, C . albicans null mutants that failed to secrete phospholipase B, encoded by caPLB1, were constructed . When these isogenic strain pairs were tested in two clinically relevant murine models of candidiasis, deletion of caPLB1 was shown to lead to attenuation of candidal virulence . Importantly, immunogold electron microscopy studies showed that C . albicans secretes this enzyme during the infectious process . These data indicate that phospholipase B is essential for candidal virulence . Although the mechanism(s) through which phospholipase modulates fungal virulence is still under investigations, early data suggest that direct host cell damage and lysis are the main mechanisms contributing to fungal virulence . Since the importance of phospholipases in fungal virulence is already known, the next challenge will be to utilize these lytic enzymes as therapeutic and diagnostic targets.

Clin Infect Dis, 2000 Jan, 30(1), 47 - 54
Diagnosis and management of increased intracranial pressure in patients with AIDS and cryptococcal meningitis . The NIAID Mycoses Study Group and AIDS Cooperative Treatment Groups; Graybill JR et al.; This study was undertaken to characterize the laboratory and clinical course of patients with AIDS and cryptococcal meningitis who had normal or elevated cerebrospinal fluid (CSF) pressure . Data were obtained retrospectively from a randomized multicenter quasifactorial phase III study comparing amphotericin B with or without flucytosine in primary treatment of cryptococcal meningitis . CSF pressure was measured before treatment and at 2 weeks . Repeated lumbar punctures were done to drain CSF and to reduce pressure . Patients with the highest baseline opening pressures (> or = 250 mm H2O) were distinguished by higher titers of cryptococcal capsular polysaccharide antigen in CSF; more frequently positive India ink smears of CSF; and more frequent headache, meningismus, papilledema, hearing loss, and pathological reflexes . After receiving antifungal therapy, those patients whose CSF pressure was reduced by >10 mm or did not change had more frequent clinical response at 2 weeks than did those whose pressure increased >10 mm (P<.001) . Patients with pretreatment opening pressure <250 mm H2O had increased short-term survival compared with those with higher pressure . We recommend that opening pressures >/=250 mm H2O be treated with large-volume CSF drainage.

Clin Infect Dis, 2000 Jan, 30(1), 29 - 34
Disseminated infection due to rapidly growing mycobacteria in immunocompetent hosts presenting with chronic lymphadenopathy: a previously unrecognized clinical entity; Chetchotisakd P et al.; Disseminated infection due to rapidly growing mycobacteria is uncommon and occurs mostly in immunocompromised patients . We report 16 cases of such infection with an unusual presentation seen at Srinagarind Hospital, a university hospital in northeastern Thailand . The clinical features were different from those in previous reports . All of the patients presented with chronic bilateral cervical lymphadenopathy . Twelve had mycobacterial involvement of other organs (sinuses, 6 patients; lungs, 4; liver, 4; spleen, 3; skin, 3; bone and joint, 2; and tonsils, 2) . An interesting occurrence in 11 patients was 14 episodes of reactive skin manifestations (Sweet's syndrome, 9; generalized pustulosis and erythema nodosum, 2 each; and pustular psoriasis, 1) . No identifiable predisposing factors, including human immunodeficiency disease, were found in these patients . However, 8 patients had 11 episodes of prior infection or coinfection with other opportunistic pathogens (salmonellosis, 4; penicilliosis, 3; pulmonary tuberculosis, 2; and melioidosis and cryptococcosis, 1 each) . These findings suggest that cell-mediated immunity is defective in these patients.

Clin Diagn Lab Immunol, 2000 Jan, 7(1), 125 - 8
Development of positive selectable markers for the fungal pathogen Cryptococcus neoformans; Hua J et al.; Cryptococcus neoformans is an opportunistic fungal pathogen that causes meningitis in approximately 10% of patients with AIDS . New selectable markers which confer resistance to G418 or phleomycin when transformed into C . neoformans were made . A hygromycin-selectable marker was modified to allow selection with a single copy of the marker.

Gastroenterol Clin Biol, 1999 Nov, 23(11), 1251 - 3
{Late cryptococcal meningitis after liver transplantation}; Dumortier J et al.; We report a case of cryptococcal meningitis, eight years after liver transplantation for primary biliary cirrhosis . Detection of the cryptococcal antigen in serum and cerebrospinal fluid appears to be essential for initial diagnosis and follow-up . Oral fluconazole treatment alone can be effective, when given for a very long period to prevent relapse.

Am J Med Sci, 1999 Dec, 318(6), 419 - 23
Hypercalcemia associated with infection by Cryptococcus neoformans and Coccidioides immitis; Ali MY et al.; BACKGROUND: Of the 13 reported cases of hypercalcemia associated with fungal infection, 1 was caused by Cryptococcus neoformans and probably mediated by increased levels of 1,25-dihydroxyvitamin D {1,25(OH)2D} . Eight others were associated with Coccidioides immitis, of which only 2 had measured 1,25(OH)2D levels; in both, they were diminished . We report a patient with human immunodeficiency virus infection and simultaneous C . immitis and C . neoformans pneumonia and C . immitis fungemia associated with hypercalcemia . METHODS: Consecutive measurements of serum total and ionized calcium, phosphorous, blood urea nitrogen, creatinine, 25(OH)D, 1,25(OH)2D, parathyroid hormone (PTH), parathyroid hormone-related protein (PTHrp) and albumin were performed over a period of 46 months . RESULTS: While the patient was hypercalcemic, intact serum PTH and PTHrp were undetectable, serum 25(OH)D levels were normal, and serum 1,25(OH)2D levels were in the high normal range . Successful treatment of the C . immitis and C . neoformans infections resulted in resolution of the hypercalcemia and increase of PTH and PTHrp to the normal range . CONCLUSION: In some patients with HIV infection, coincident hypercalcemia, and severe fungal infection, the responsible factor may be 1,25(OH)2D . Although total serum levels of this compound may not be frankly elevated, they are inappropriately high for the circumstances . Free 1,25(OH)2D levels should be determined in this situation.

Rev Invest Clin, 1999 Sep-Oct, 51(5), 303 - 7
{Significance of intracranial hypertension management in cryptococcal meningitis in patients with acquired immunodeficiency syndrome . Report of 2 cases}; Nino Oberto S et al.; Two cases of cryptococcal meningitis and increased intracranial pressure in patients with acquired immunodeficiency are described . Both patients presented high intracranial pressure that persisted despite optimal antifungal treatment (amphotericin B, 5-flucytosine initially, and fluconazole posteriorly) . The elevated intracranial pressure produced headache, seizures, and reduced visual and auditory acuity . CAT scan demonstrated absence of ventricular dilatation or focal lesions . Both cases were treated with adequate antifungal therapy, as well as with repeated lumbar punctures and placement of a lumboperitoneal shunt due to the persistence of elevated intracranial pressure . One patient presented with unilateral loss of vision due to optic nerve atrophy . After one year of follow-up, one patient died due to progression of his disease, while the other is still alive and without evidence of neurological disease . Intracranial hypertension is a frequent clinical manifestation of cryptococcal meningitis in patients with acquired immunodeficiency syndrome (AIDS) that requires adequate diagnosis and management . Treatment should be directed towards the reduction of intracranial pressure though repeated lumbar punctures and, in some cases, with lumboperitoneal or ventricular-peritoneal shunts.

Microbes Infect, 1999 Apr, 1(4), 293 - 301
Pathogenicity of Cryptococcus neoformans: virulence factors and immunological mechanisms; Rodrigues ML et al.; Cryptococcus neoformans is the causative agent of cryptococcosis and cryptococcal meningitis, which are serious pathological conditions affecting up to 10% of patients with AIDS . Mechanisms of pathogenicity of C . neoformans and the host defenses against this fungus are reviewed, incorporating recent data and perspectives.

Chem Pharm Bull (Tokyo), 1999 Nov, 47(11), 1591 - 7
Tetranorditerpene lactones, potent antifungal antibiotics for human pathogenic yeasts, from a unique species of Oidiodendron; Hosoe T et al.; The culture filtrate of a fungus isolated from decaying Picea glauca wood and tentatively identified as Oidiodendron cf . truncatum showed strong antibiotic activity against the pathogenic yeast, Candida albicans . Four new tetranorditerpenoids, oidiodendrolides A (3), B (4), and C (5) and oidiodendronic acid (7) were isolated along with three known tetranorditerpenoids, LL-Z1271 alpha (= PR1387) (1), PR1388 (2), and acrostalidic acid (6), from rice fermented by the above fungus . The structures of oidiodendrolides A (3), B (4), and C (5) and oidiodendronic acid (7) were established on the basis of spectroscopic and chemical investigations . The antifungal activity of the above tetranorditerpenoids against the pathogenic yeasts, Candida albicans and Cryptococcus neoformans is discussed.

Antimicrob Agents Chemother, 2000 Jan, 44(1), 226 - 9
In vitro activities of the new antifungal triazole SCH 56592 against common and emerging yeast pathogens; Barchiesi F et al.; A broth microdilution method performed in accordance with the National Committee for Clinical Laboratory Standards guidelines was used to compare the in vitro activity of the new antifungal triazole SCH 56592 (SCH) to that of fluconazole (FLC), itraconazole (ITC), and ketoconazole (KETO) against 257 clinical yeast isolates . They included 220 isolates belonging to 12 different species of Candida, 15 isolates each of Cryptococcus neoformans and Saccharomyces cerevisiae, and seven isolates of Rhodotorula rubra . The MICs of SCH at which 50% (MIC(50)) and 90% (MIC(90)) of the isolates were inhibited were 0.06 and 2.0 microg/ml, respectively . In general, SCH was considerably more active than FLC (MIC(50) and MIC(90) of 1.0 and 64 microg/ml, respectively) and slightly more active than either ITC (MIC(50) and MIC(90) of 0.25 and 2.0 microg/ml, respectively) and KETO (MIC(50) and MIC(90) of 0.125 and 4.0 microg/ml, respectively) . Our in vitro data suggest that SCH has significant potential for clinical development.

Antimicrob Agents Chemother, 2000 Jan, 44(1), 143 - 9
Immunosuppressive and nonimmunosuppressive cyclosporine analogs are toxic to the opportunistic fungal pathogen Cryptococcus neoformans via cyclophilin-dependent inhibition of calcineurin; Cruz MC et al.; Cyclosporine (CsA) is an immunosuppressive and antimicrobial drug which, in complex with cyclophilin A, inhibits the protein phosphatase calcineurin . We recently found that Cryptococcus neoformans growth is resistant to CsA at 24 degrees C but sensitive at 37 degrees C and that calcineurin is required for growth at 37 degrees C and pathogenicity . Here CsA analogs were screened for toxicity against C . neoformans in vitro . In most cases, antifungal activity was correlated with cyclophilin A binding in vitro and inhibition of the mixed-lymphocyte reaction and interleukin 2 production in cell culture . Two unusual nonimmunosuppressive CsA derivatives, (gamma-OH) MeLeu(4)-Cs (211-810) and D-Sar (alpha-SMe)(3) Val(2)-DH-Cs (209-825), which are also toxic to C . neoformans were identified . These CsA analogs inhibit C . neoformans via fungal cyclophilin A and calcineurin homologs . Our findings identify calcineurin as a novel antifungal drug target and suggest nonimmunosuppressive CsA analogs warrant investigation as antifungal agents.

Antimicrob Agents Chemother, 2000 Jan, 44(1), 57 - 62
In vitro activities of a new lipopeptide antifungal agent, FK463, against a variety of clinically important fungi; Tawara S et al.; The in vitro antifungal activity and spectrum of FK463 were compared with those of amphotericin B, fluconazole, and itraconazole by using a broth microdilution method specified by National Committee for Clinical Laboratory Standards document M27-A (National Committee for Clinical Laboratory Standards, Wayne, Pa., 1997) . FK463 exhibited broad-spectrum activity against clinically important pathogens including Candida species (MIC range, <==0.0039 to 2 microg/ml) and Aspergillus species (MIC range, <==0.0039 to 0.0313 microg/ml), and its MICs for such fungi were lower than those of the other antifungal agents tested . FK463 was also potently active against azole-resistant Candida albicans as well as azole-susceptible strains, and there was no cross-resistance with azoles . FK463 showed fungicidal activity against C . albicans, i.e., a 99% reduction in viability after a 24-h exposure at concentrations above 0.0156 microg/ml . The minimum fungicidal concentration (MFC) assays indicated that FK463 was fungicidal against most isolates of Candida species . In contrast, the MFCs of FK463 for A . fumigatus isolates were much higher than the MICs, indicating that its action is fungistatic against this species . FK463 had no activity against Cryptococcus neoformans, Trichosporon species, or Fusarium solani . Neither the test medium (kind and pH) nor the inoculum size greatly affected the MICs of FK463, while the addition of 4% human serum albumin increased the MICs for Candida species and A . fumigatus more than 32 times . Results from preclinical in vitro evaluations performed thus far indicate that FK463 should be a potent parenteral antifungal agent.

FEBS Lett, 1999 Dec 10, 463(1-2), 58 - 62
SMAP-29: a potent antibacterial and antifungal peptide from sheep leukocytes; Skerlavaj B et al.; SMAP-29 is a cathelicidin-derived peptide deduced from sheep myeloid mRNA . The C-terminally amidated form of this peptide was chemically synthesized and shown to exert a potent antimicrobial activity . Antibiotic-resistant clinical isolates highly susceptible to this peptide include MRSA and VREF isolates, that are a major worldwide problem, and mucoid Pseudomonas aeruginosa associated with chronic respiratory inflammation in CF patients . In addition, SMAP-29 is also active against fungi, including Cryptococcus neoformans isolated from immunocompromised patients . SMAP-29 causes significant morphological alterations of the bacterial surfaces, as shown by scanning electron microscopy, and is also hemolytic against human, but not sheep erythrocytes . Its potent antimicrobial activity suggests that this peptide is an excellent candidate as a lead compound for the development of novel antiinfective agents.

J Microbiol Immunol Infect, 1998 Jun, 31(2), 101 - 6
Bacteremia and fungemia in hematological and oncological children with neutropenic fever: two-year study in a medical center; Chiu HH et al.; A retrospective study of bacteremia in children with neutropenic fever admitted to a medical center in Taiwan from Jan . 1994 to Dec . 1995 was performed . There were in total 273 episodes of neutropenic fever during this period, but only 79 pathogens were isolated from blood specimens in 70 episodes . Klebsiella pneumoniae (27.8%), E . coli (10.1%), Staphylococcus aureus (10.1%) and Pseudomonas aeruginosa (7.6%) were the most common pathogens . All the isolates of S . aureus were methicillin sensitive . About half of K . pneumoniae (10/22) was multiple-drug resistant . There were seven infection-related mortality cases, three due to multiple-drug resistant K . pneumoniae, one due to S . aureus, one alpha-hemolytic streptococcus and two fungemia (Cryptococcus neoformans and Fusarium sp.) . Vancomycin is not necessary in initial empiric therapy of neutropenic fever, while cefazolin or oxacillin may be included in cases with central venous access device . Antibiotics to cover intestinal flora, especially K . pneumoniae, are paramount in our hospital.

Ther Umsch, 1999 Nov, 56(11), 670 - 4
{CNS-infections in HIV patients}; Malinverni R et al.; Neurological manifestations are frequent in patients with AIDS . Many neurological disorders have disappeared with the advent of highly active antiretroviral combination therapies . We can speculate that some of these disorders may reappear in patients under antiretroviral therapy, possibly with different clinical manifestations and at a different stage during HIV-infection . We discuss the appearance of the most common neurological complications in relation to the CD4-cell count during HIV-infection . The most frequent causes of seizures and headache in HIV-infected patients are shown . We recommend a systematic diagnostic work-up in patients with headache, starting from 3 typical clinical situations: focal signs, convulsions or altered mental status; no focal signs, CD4-cells > 200 microliters, meningism; fever and/or meningism, no focal signs . The analysis of the cerebrospinal fluid by polymerase chain reaction is now a well established diagnostic method for investigating the most common CNS-infections in AIDS-patients . Neuroimaging (by MRI or CT-scan) is an additional, useful investigation . Cerebral toxoplasmosis, cryptococcosis, PML, encephalitis due to herpes-viruses and neurosyphilis are discussed.

J Am Vet Med Assoc, 1999 Nov 1, 215(9), 1301 - 5
Endoscopic examination of the choanae in dogs and cats: 118 cases (1988-1998); Willard MD et al.; OBJECTIVE: To determine whether endoscopic examination of the choanae resulted in diagnosis of various diseases in dogs and cats with signs of respiratory tract disease . DESIGN: Retrospective study . ANIMALS: 91 dogs and 27 cats that had endoscopic examination of the choanae . PROCEDURE: Medical records were reviewed for endoscopy findings and results of examination of biopsy or cytologic specimens . RESULTS: 34 animals had neoplasia in the choanal region; in 26 animals, diagnosis was confirmed by evaluation of specimens obtained by endoscopy . Five dogs with neoplasia had an erroneous diagnosis of rhinitis made on the basis of evaluation of specimens obtained by endoscopy . Six dogs and 2 cats had foreign objects in the choanae; 7 foreign objects were removed endoscopically, whereas 1 required nasal flushing . Results of endoscopy and biopsy of the choanae provided diagnosis of cryptococcosis and aspergillosis, but did not aid in the diagnosis of pythiosis or nasal mites . CONCLUSIONS AND CLINICAL RELEVANCE: Endoscopic examination of the choanae may assist in rapid diagnosis of nasal neoplasms, foreign objects, and certain infectious organisms.

Mol Cell Biol, 2000 Jan, 20(1), 352 - 62
The G-protein beta subunit GPB1 is required for mating and haploid fruiting in Cryptococcus neoformans; Wang P et al.; Cryptococcus neoformans is an opportunistic fungal pathogen with a defined sexual cycle . The gene encoding a heterotrimeric G-protein beta subunit, GPB1, was cloned and disrupted . gpb1 mutant strains are sterile, indicating a role for this gene in mating . GPB1 plays an active role in mediating responses to pheromones in early mating steps (conjugation tube formation and cell fusion) and signals via a mitogen-activated protein (MAP) kinase cascade in both MATalpha and MATa cells . The functions of GPB1 are distinct from those of the Galpha protein GPA1, which functions in a nutrient-sensing cyclic AMP (cAMP) pathway required for mating, virulence factor induction, and virulence . gpb1 mutant strains are also defective in monokaryotic fruiting in response to nitrogen starvation . We show that MATa cells stimulate monokaryotic fruiting of MATalpha cells, possibly in response to mating pheromone, which may serve to disperse cells and spores to locate mating partners . In summary, the Gbeta subunit GPB1 and the Galpha subunit GPA1 function in distinct signaling pathways: one (GPB1) senses pheromones and regulates mating and haploid fruiting via a MAP kinase cascade, and the other (GPA1) senses nutrients and regulates mating, virulence factors, and pathogenicity via a cAMP cascade.

J Antimicrob Chemother, 1999 Dec, 44(6), 827 - 9
In-vitro and in-vivo activities of SCH56592 against Cryptococcus neoformans; Hossain MA et al.; The in-vitro and in-vivo activities of SCH56592, a triazole antifungal agent, against Cryptococcus neoformans were studied . MIC(90)s for 16 strains of C . neoformans measured by microdilution method (NCCLS M27-A) were 1 mg/L of SCH56592, 16 mg/L of fluconazole, 32 mg/L of flucytosine, and 0.5 mg/L of amphotericin B . In a murine model of pulmonary cryptococcosis, 10 mg/kg of SCH56592 was more effective than fluconazole . The fungal burden of the lung of animals treated with SCH56592 was significantly reduced (7.40 +/- 0.21 log(10) cfu/g), as compared with fluconazole (7.77 +/- 0.07 log(10) cfu/g) and control (7.79 +/- 0.1 log(10) cfu/g) (P < 0.01) . For C . neoformans-infected mice following 7 days treatment with 10 mg/kg of SCH56592 there was a higher concentration in lung (3.36 +/- 0.62 ng/ml) than in plasma (2.16 +/- 0.86 ng/mL), and this was maintained for 12 h after administration.

Mycoses, 1999, 42(9-10), 543 - 8
Fungaemia survey: a 10-year experience in Bergamo, Italy; Farina C et al.; The authors analysed 10 years of experience of fungaemia at a Regional Italian Hospital, the Azienda Ospedaliera (A.O.) 'Ospedali Riuniti di Bergamo', Bergamo, Italy, from 1988 to 1997 . One hundred and sixty-eight cases were observed, with a global incidence corresponding to 3.43/10,000 in-patients . Median age was 38.5 years and mean age was 38.9 years (range: 0-94 years) . Female:male ratio was 1:1.75 . Fungaemia occurred 25.7 days (mean value) after admission to the hospital . Aetiology was: 134 Candida spp . (70%), 11 Cryptococcus neoformans (6.5%), seven Torulopsis inconspicua (4.1%), three Trichosporon beigelii (1.8%), one Hansenula anomala (0.6%); three Fusarium verticillioides (1.8%), three Geotrichum candidum (1.8%) and one Histoplasma capsulatum (0.6%) . Total mortality was 50.6%, and particularly related to Candida kefyr and Candida krusei, to Cr . neoformans and Fusarium spp.

Antimicrob Agents Chemother, 1999 Dec, 43(12), 2862 - 8
Comparison of in vitro activities of camptothecin and nitidine derivatives against fungal and cancer cells; Del Poeta M et al.; The activities of a series of camptothecin and nitidine derivatives that might interact with topoisomerase I were compared against yeast and cancer cell lines . Our findings reveal that structural modifications to camptothecin derivatives have profound effects on the topoisomerase I-drug poison complex in cells . Although the water-soluble anticancer agents topotecan and irinotecan are less active than the original structure, camptothecin, other derivatives or analogs with substitutions that increase compound solubility have also increased antifungal activities . In fact, a water-soluble prodrug appears to penetrate into the cell and release its active form; the resulting effect in complex with Cryptococcus neoformans topoisomerase I is a fungicidal response and also potent antitumor activity . Some of the compounds that are not toxic to wild-type yeast cells are extremely toxic to the yeast cells when the C . neoformans topoisomerase I target is overexpressed . With the known antifungal mechanism of a camptothecin-topoisomerase I complex as a cellular poison, these findings indicate that drug entry may be extremely important for antifungal activity . Nitidine chloride exhibits antifungal activity against yeast cells through a mechanism(s) other than topoisomerase I and appears to be less active than camptothecin analogs against tumor cells . Finally, some camptothecin analogs exhibit synergistic antifungal activity against yeast cells in combination with amphotericin B in vitro . Our results suggest that camptothecin and/or nitidine derivatives can exhibit potent antifungal activity and that the activities of camptothecin derivatives with existing antifungal drugs may be synergistic against pathogenic fungi . These new compounds, which exhibit potent antitumor activities, will likely require further structural changes to find more selective activity against fungal versus mammalian cells to hold promise as a new class of antifungal agents.

Microbios, 1999, 100(395), 27 - 40
Indigenous yeasts associated with two Vitis vinifera grape varieties cultured in southern Spain; De La Torre MJ et al.; The yeast microbiota present on the surface of grapes of two Vitis vinifera varieties, Pedro Ximenez and Tempranillo de Rioja, grown in the Montilla-Moriles region of southern Spain was identified . The changes between veraison and the physiological ripeness time during 3 years were monitored . Overall, the yeast microbiota isolated was of oxidative metabolism . Sporobolomyces roseus and Cryptococcus albidus species occurred at all physiological stages, in the two Vitis vinifera varieties, and the three seasons studied . On the other hand, Kloeckera apiculata was never detected and Saccharomyces cerevisiae was scarcely isolated, it was only present, testimonial, in Tempranillo de Rioja grapes during the 1992 vintage . The widest variety of yeast species was observed in the 1992 season, and in contrast, the lowest number of species in both varieties of Vitis was detected in the 1994 season.

Genetics, 1999 Dec, 153(4), 1601 - 15
The STE12alpha homolog is required for haploid filamentation but largely dispensable for mating and virulence in Cryptococcus neoformans; Yue C et al.; Cryptococcus neoformans is a fungal pathogen that causes meningitis in immunocompromised hosts . The organism has a known sexual cycle, and strains of the MATalpha mating type are more virulent than isogenic MATa strains in mice, and they are more common in the environment and infected hosts . A C . neoformans homolog of the STE12 transcription factor that regulates mating, filamentation, and virulence in Saccharomyces cerevisiae and Candida albicans was identified previously, found to be encoded by a novel region of the MATalpha mating type locus, and shown to enhance filamentous growth when overexpressed . We have disrupted the C . neoformans STE12 gene in a pathogenic serotype A isolate . ste12 mutant strains exhibit a severe defect in filamentation and sporulation (haploid fruiting) in response to nitrogen starvation . In contrast, ste12 mutant strains have only modest mating defects and are fully virulent in two animal models compared to the STE12 wild-type strain . In genetic epistasis experiments, STE12 functions in a MAP kinase cascade to regulate fruiting, but not mating . Thus, the C . neoformans STE12alpha transcription factor homolog plays a specialized function in haploid fruiting, but it is dispensable or redundant for mating and virulence . The association of the MATalpha locus with virulence may involve additional genes, and other transcription factors that regulate mating and virulence remain to be identified.

J Antibiot (Tokyo), 1999 Aug, 52(8), 695 - 9
Rhodopeptins (Mer-N1033), novel cyclic tetrapeptides with antifungal activity from Rhodococcus sp . I . Taxonomy, fermentation, isolation, physico-chemical properties and biological activities; Chiba H et al.; Five novel cyclic tetrapeptides, named rhodopeptin C1, C2, C3, C4 and B5, were isolated from a strain named Rhodococcus sp . Mer-N1033 . They are a novel type of cyclic tetrapeptide composed of a beta-amino acid and three usual alpha-amino acids . Rhodopeptins show high in vitro antifungal activity against Candida albicans and Cryptococcus neoformans, whereas they show no activity against bacteria.

FEMS Immunol Med Microbiol, 1999 Dec, 26(3-4), 309 - 18
Cryptococcus neoformans and its cell wall components induce similar cytokine profiles in human peripheral blood mononuclear cells despite differences in structure; Walenkamp AM et al.; We studied the cytokine profile of peripheral blood mononuclear cells after stimulation with various cryptococcal strains or its purified cell wall components . After 3 h of stimulation, tumor necrosis factor (TNF) alpha levels were strongly increased, whereas interferon (IFN) gamma and interleukin (IL) 10 levels were increased only slightly, or not at all (respectively) . In contrast, after 18 h, TNF-alpha and IFN-gamma levels were (strongly) decreased, whereas the IL-10 levels were increased . The IL-1beta, IL-6 and IL-8 levels were equally high throughout the experiment . In order to establish which of the cryptococcal envelope components contributed most to the observed cytokine profile induced by whole cryptococci, glucuronoxylomannan, galactoxylomannan and mannoproteins were purified and partially characterized biochemically . All cryptococcal components elicited a similar cytokine pattern despite the differences in structure.

Yeast, 1999 Nov, 15(15), 1657 - 67
Inositol synthesis and catabolism in Cryptococcus neoformans; Molina Y et al.; Cryptococcus neoformans is an opportunistic fungal pathogen that synthesizes and catabolizes inositol . This study demonstrates inositol synthesis from glucose-6-phosphate via inositol-1-phosphate synthase and catabolism to glucuronic acid via inositol oxygenase in this organism . These inositol synthetic and catabolic pathways are regulated in opposition; repressing conditions for one are inducing conditions for the other . An inositol-requiring strain was generated by UV mutagenesis . Without inositol, this mutant strain undergoes 'inositol-less' death, during which time the phosphatidylinositol composition of the membranes decreases without alteration of the proportion of other phospholipids . The mutation on this strain results in no detectable inositol synthetic activity but normal (wild-type) inositol catabolic activity . This inositol-requiring mutant strain reverted at a high frequency . Classical genetic experiments revealed that the majority of the reverting mutations are at second sites . Interestingly, the revertants exhibited unusual morphological phenotypes when deprived of inositol, while provision of inositol restored wild-type morphology . Inositol metabolism is clearly important for growth and development of C . neoformans and may be involved in this organism's mechanism for survival as both a saprophyte in soil and a parasite in humans .

Appl Microbiol Biotechnol, 1999 Oct, 52(4), 495 - 501
Production of sophorolipids from whey . II . Product composition, surface active properties, cytotoxicity and stability against hydrolases by enzymatic treatment; Otto RT et al.; Sophorolipids, obtained by a two-stage process starting from deproteinized whey concentrate using Cryptococcus curvatus ATCC 20509 and Candida bombicola ATCC 22214, were compared to products from one-stage processes, using different lipidic compounds as substrates . Results showed that above all carbon source and not cultivation conditions had a distinct influence on the composition of the crude product mixture and therefore on the physicochemical and biological properties of the sophorolipids, such as, for example, surface activity, cytotoxicity and stability against hydrolases . The results were completed by corresponding data for purified mono- and diacetylated (17-hydroxyoctadecenoic)-1',4"-lactonized sophorolipids . Crude sophorolipid mixtures showed moderate to good surface active properties (SFTmin 39 mN m-1, CMC 130 mg l-1), water solubilities (2-3 g l-1) and low cytotoxicities (LC50 300-700 mg l-1) . In contrast, purified sophorolipids were more surface active (SFTmin 36 mN m-1, CMC 10 mg l-1), less water soluble (max . 70 mg l-1) and showed stronger cytotoxic effects (LC50 15 mg l-1) . Incubation of crude sophorolipid mixtures with different hydrolases demonstrated that treatment with commercially available lipases such as from Candida rugosa and Mucor miehei distinctly reduced the surface active properties of the sophorolipids, while treatment with porcine liver esterase and glycosidases had no effect.

Rev Esp Quimioter, 1999 Jun, 12(2), 126 - 35
{Determination of the in vitro antifungal susceptibility of clinically important yeasts using the Sensititre system}; Carrillo-Munoz AJ et al.; Using Sensititre (AccuMed, USA) we studied the in vitro antifungal activity of amphotericin B, fluconazole, itraconazole, ketoconazole and 5-fluorocytosine against 250 clinical yeast isolates taken from different hospitals, including Candida (151 C . albicans, 15 C . krusei, 14 C . parapsilosis, 11 C . tropicalis, 10 C . glabrata, 4 C . guilliermondii, 3 C . rugosa, 2 C . viswanathii, 2 C . famata and 2 C . kefyr), Cryptococcus (32 C . neoformans and 1 C . laurentii), Trichosporon (2 isolates) and Rhodotorula rubra (1 isolate) . All the strains were susceptible to amphotericin B and showed an MIC <1 mg/l . The susceptibility of C . albicans (MIC(90) <256 mg/l), C . krusei (MIC(90) <64 mg/l), C . glabrata (MIC(90) <64 mg/l) and C . neoformans (MIC(90) 32 mg/l) to fluconazole was lower (14% isolates being resistant and 16.8% susceptible depending on the dose) . The largest number of strains resistant to itraconazole was observed in C . albicans and C . glabrata (17.2% resistant and 24% susceptible and susceptible depending on the dose, respectively) . Ketoconazole and 5-fluorocytosine were not effective in vitro against 12.8% and 2%, respectively, of all the isolates studied . Nine C . krusei and seven C . neoformans (12.9%) showed dose-dependent susceptibility to 5-fluorocytosine.

Infect Immun, 1999 Dec, 67(12), 6314 - 20
Cytokine profiles of AIDS patients are similar to those of mice with disseminated Cryptococcus neoformans infection; Lortholary O et al.; Cryptococcosis is an hematogenously disseminated meningoencephalitis during which the relationship between the disease severity and the immune response remains unclear . We thus analyzed, by enzyme-linked immunosorbent assay, proinflammatory (tumor necrosis factor alpha {TNF-alpha} and interleukin-6 {IL-6}) and anti-inflammatory (IL-10) cytokine levels in plasma at the time of diagnosis in 51 AIDS patients with culture-proven cryptococcosis . We used a murine model to determine the correlation between cytokine levels and fungal burden in blood and tissues and the kinetics of the immune response and of the formation of cerebral lesions . In AIDS patients, plasma TNF-alpha and IL-10, but not IL-6, levels were significantly higher in the case of fungemia or disseminated infection than in their absence, whereas the presence of meningitis had no influence on these levels . In mice, none of these cytokines were detected within the first day after inoculation . Later on, TNF-alpha and IL-10, but not IL-6, levels in plasma correlated significantly with the fungal burden in the blood and spleen but not the brain . In the brain, cytokine levels were low compared to those in other compartments, and tissue lesions and a degree of infection similar to those observed in humans were seen, further suggesting the relevance of this experimental model . Thus, AIDS patients with cryptococcosis produce an immune response that reflects the dissemination but not the meningeal involvement . This murine model of disseminated cryptococcosis can be used to investigate the pathophysiology of cryptococcosis and new therapeutic approaches.

AIDS, 1999 Nov 12, 13(16), 2197 - 207
HIV type 1 envelope glycoprotein gp120 induces development of a T helper type 2 response to Cryptococcus neoformans; Pietrella D et al.; OBJECTIVE: To analyse the contribution of HIV type 1 envelope glycoprotein gp120 to regulation of a T-cell response to Cryptococcus neoformans . DESIGN: Monocytes treated with recombinant gp120 and exposed to C . neoformans were used as antigen presenting cells (APC) in coculture with autologous T lymphocytes . METHODS: Costimulatory and major histocompatibility complex class II molecules were evaluated on APC by flow cytometry analysis . T-cell proliferation was determined as 3H thymidine incorporation . Cytokine production was analysed by enzyme-linked immunosorbent assay . RESULTS: gp120 had multiple effects on APC and the T-cell response including: (i) up-regulation of major histocompatibility complex class II antigens on the APC surface resulting from both redistribution of molecules from the intracellular pool and synthesis of new molecules; (ii) up-regulation of B7-2 molecules on the APC surface; (iii) altered T-cell proliferation; and (iv) promotion of interleukin-4 and inhibition of interferon-gamma synthesis and release . CONCLUSIONS: These data indicate that gp120 alters the normal T-cell response to C . neoformans, promoting a T-helper type 2 response . The altered T-cell response produced by gp120 may play an important role in the pathogenesis of cryptococcosis in the patient with AIDS.

J Med Assoc Thai, 1999 Oct, 82(10), 991 - 9
Successful medical treatment of multiple cryptococcomas: report of a case and literature review; Arayawichanont A et al.; We report a 35-year-old man diagnosed as having CNS cryptococcosis with multiple cryptococcomas, presenting with headache, papilloedema and impaired mental function in a previously healthy man . Cerebrospinal fluid (CSF) examination revealed lymphocytic pleocytosis with low glucose level . Gram's stain, acid fast bacilli stain and Indian ink examination were all negative . CSF cryptococcal antigen was positive, however, several fungal cultures were negative . Early cranial CT scan showed focal cerebritis over the right temporal lobe while subsequent imaging studies showed multiple contrast-enhancing masses with severe surrounding brain oedema over bilateral frontoparietal areas . Brain biopsy showed cryptococcal granulomatous lesions . Treatment was successful with antifungal agents and steroids without surgical removal.

J Formos Med Assoc, 1999 Sep, 98(9), 621 - 6
Pulmonary cryptococcosis: manifestations in the era of acquired immunodeficiency syndrome; Wu TT et al.; To examine the clinical manifestations, treatment, and outcome of pulmonary cryptococcosis, we reviewed the medical records of all patients treated for Cryptococcus neoformans infection at our hospital from January 1988 through September 1998 . Sixty-three patients were included in the analysis, 10 (16%) of whom had acquired immunodeficiency syndrome (AIDS) . Thirty-four of the 53 non-AIDS patients, including 19 men and 15 women had pulmonary cryptococcosis, including 31 with isolated pulmonary cryptococcosis and three with disseminated disease . Of the 10 AIDS patients, seven presented with disseminated cryptococcosis (including one patient with lung involvement) and one had isolated cryptococcal lung disease . The age (mean +/- SD) of the 34 non-AIDS patients with pulmonary cryptococcosis was 52.1 +/- 15.2 years (range, 19-75 yr) . Cough was the most common symptom (58%) . Diabetes mellitus (12%) and malignancy (12%) were two major underlying diseases . Nodules and masses were the predominant manifestations of pulmonary cryptococcosis in non-AIDS patients (79%) . The most frequently used diagnostic modality for pulmonary cryptococcosis was biopsy with/without aspiration under ultrasound guidance (56%) . Antifungal therapy (20/34) was the most common treatment for non-AIDS patients, followed by surgical resections with antifungal therapy (9), surgical resections alone (3), and no treatment (2) . Antifungal therapy and/or resection yielded excellent outcomes (total recovery, 27; improvement, 4) . Of the 18 patients who underwent lumbar puncture, only two had positive cerebrospinal fluid (CSF) cultures for C . neoformans, both had symptoms and signs of increased intracranial pressure . There was no clinical evidence of meningitis in the other 32 patients . Our findings indicate that pulmonary cryptococcosis in non-AIDS patients tends to be a more localized and benign process than in AIDS patients . Ultrasound-guided lung biopsy or aspiration is an effective tool for diagnosis . CSF examination may not be mandatory as an initial routine procedure for pulmonary cryptococcosis in non-AIDS patients.

Dermatology, 1999, 199(2), 180 - 2
Cryptococcosis during systemic glucocorticosteroid treatment; Lauerma AI et al.; Cryptococcosis is an opportunistic infection caused by a fungus, Cryptococcus neoformans . It is usually seen in immunocompromised patients with AIDS, leukaemia, lymphoma, sarcoidosis or immunosuppressive treatments . We describe a patient who was treated with systemic glucocorticosteroids for 4 years because of lung sarcoidosis . During the last year of treatment, a papular eruption developed which later became ulcerative . In a histopathological examination of a skin biopsy, there was granulomatous inflammation, and the disease was treated as sarcoidosis without success . After 1 year's unsuccessful treatment, another skin biopsy and skin fungal culture revealed C . neoformans . Cryptococcal antigen was found in blood and cerebrospinal fluid, too . The patient was successfully treated first with an amphotericin-B-flucytosine combination and later with fluconazole.

Cell Immunol, 1999 Oct 10, 197(1), 55 - 61
Interleukin-4 weakens host resistance to pulmonary and disseminated cryptococcal infection caused by combined treatment with interferon-gamma-inducing cytokines; Kawakami K et al.; We examined the role of interleukin (IL)-4 in host resistance against infection with Cryptococcus neoformans . First, we examined the effects of a neutralizing anti-IL-4 monoclonal antibody (mAb) on survival of mice infected intratracheally with this fungal pathogen . We also compared the number of live C . neoformans in lungs and brains of treated and untreated mice . Treatment with anti-IL-4 mAb significantly prolonged survival of infected mice and reduced the lung and brain burdens of C . neoformans, which was associated with increased production of IFN-gamma in lungs . In the next experiments, infected mice were treated with two IFN-gamma-inducing cytokines, IL-12 and IL-18, known to enhance protection against infection . We then evaluated the effect of such treatment on the number of live microorganisms and concentration of IL-4 in lungs . These two parameters showed a statistically significant relationship, suggesting a negative regulation of host protection by IL-4 . Finally, we examined the effects of IL-4 treatment and administration of neutralizing anti-IL-4 mAbs on host protection against C . neoformans and local production of IFN-gamma in lungs induced by treatment with IL-12/IL-18 . The former treatment suppressed host protection and reduced IFN-gamma production, while the latter produced the opposite effects . Our results indicated that IL-4 suppressed the host defense mechanisms against infection with C . neoformans potentiated by IFN-gamma-inducing cytokines probably through the suppression of local production of IFN-gamma .

Biochem J, 1999 Nov 15, 344 Pt 1, 101 - 7
Binding energy and specificity in the catalytic mechanism of yeast aldose reductases; Nidetzky B et al.; Derivatives of d-xylose and d-glucose, in which the hydroxy groups at C-5, and C-5 and C-6 were replaced by fluorine, hydrogen and azide, were synthesized and used as substrates of the NAD(P)H-dependent aldehyde reduction catalysed by aldose reductases isolated from the yeasts Candida tenuis, C . intermedia and Cryptococcus flavus . Steady-state kinetic analysis showed that, in comparison with the parent aldoses, the derivatives were reduced with up to 3000-fold increased catalytic efficiencies (k(cat)/K(m)), reflecting apparent substrate binding constants (K(m)) decreased to as little as 1/250 and, for d-glucose derivatives, up to 5.5-fold increased maximum initial rates (k(cat)) . The effects on K(m) mirror the relative proportion of free aldehyde that is available in aqueous solution for binding to the binary complex enzyme-NAD(P)H . The effects on k(cat) reflect non-productive binding of the pyranose ring of sugars; this occurs preferentially with the NADPH-dependent enzymes . No transition-state stabilization energy seems to be derived from hydrogen-bonding interactions between enzyme-NAD(P)H and positions C-5 and C-6 of the aldose . In contrast, unfavourable interactions with the C-6 group are used together with non-productive binding to bring about specificity (6-10 kJ/mol) in a series of d-aldoses and to prevent the reaction with poor substrates such as d-glucose . Azide introduced at C-5 or C-6 destabilizes the transition state of reduction of the corresponding hydrogen-substituted aldoses by approx . 4-9 kJ/mol . The total transition state stabilization energy derived from hydrogen bonds between hydroxy groups of the substrate and enzyme-NAD(P)H is similar for all yeast aldose reductases (yALRs), at approx . 12-17 kJ/mol . Three out of four yALRs manage on only hydrophobic enzyme-substrate interactions to achieve optimal k(cat), whereas the NAD(P)H-dependent enzyme from C . intermedia requires additional, probably hydrogen-bonding, interactions with the substrate for efficient turnover.

Prep Biochem Biotechnol, 1999 Nov, 29(4), 385 - 401
Preparation of novel conjugates involving immunomodulating peptidoglycan monomer; Tomasic J et al.; Peptidoglycan monomer, the disaccharide pentapeptide beta-D-Glcp-N-Ac-(1-->4)-D-Murp-N-Ac-L-Ala-D-mesoA2pm- (epsilonN H2)-D-Ala-D-Ala (PGM) is an immunomodulator . PGM and/or its derivative N-tert-butyloxycarbonyl-L-tyrosyl peptidoglycan monomer (Boc-Tyr-PGM) were coupled to two polysaccharides: the glucuronoxylomannan (GXM) from Cryptococcus neoformans, type B, solubilized by ultrasonic irradiation (MW 12-400 kDa) and to the dextran FP 70 (MW 70 kDa) . Both polysaccharides were activated by CNBr . Initially, unprotected PGM was coupled via its amino group to GXM . The reactions yielded 42%-52% of the conjugate, containing only 0.18%-0.31% of PGM . In another approach Boc-Tyr-PGM (having its amino group blocked) was reacted via its free carboxyl group . Both CNBr-activated polysaccharides were first coupled to adipic acid dihydrazide (ADH) and then subsequently coupled to Boc-Tyr-PGM . The dextran conjugate (approximately 80% yield ) contained 6.3% of Boc-Tyr-PGM . The isolation of GXM conjugate required several modifications and it was obtained in lower yield (approximately 30%) but contained 13.7% of Boc-Tyr-PGM . Both conjugates were water soluble and apyrogenic and suitable for further testing of biological activity.

Antimicrob Agents Chemother, 1999 Nov, 43(11), 2663 - 70
Accumulation of 3-ketosteroids induced by itraconazole in azole-resistant clinical Candida albicans isolates; Marichal P et al.; The effects of itraconazole on ergosterol biosynthesis were investigated in a series of 16 matched clinical Candida albicans isolates which had been previously analyzed for mechanisms of resistance to azoles (D . Sanglard, K . Kuchler, F . Ischer, J . L . Pagani, M . Monod, and J . Bille, Antimicrob . Agents Chemother., 39:2378-2386, 1995) . Under control conditions, all isolates contained ergosterol as the predominant sterol, except two strains (C48 and C56) . In isolates C48 and C56, both less susceptible to azoles than their parent, C43, substantial concentrations (20 to 30%) of 14alpha-methyl-ergosta-8,24(28)-diene-3beta,6alpha-dio l (3, 6-diol) were found . Itraconazole treatment of C43 resulted in a dose-dependent inhibition of ergosterol biosynthesis (50% inhibitory concentration, 2 nM) and accumulation of 3,6-diol (up to 60% of the total sterols) together with eburicol, lanosterol, obtusifoliol, 14alpha-methyl-ergosta-5,7,22,24(28)-tetraene-3betaol, and 14alpha-methyl-fecosterol . In strains C48 and C56, no further increase of 3,6-diol was observed after exposure to itraconazole . Ergosterol synthesis was less sensitive to itraconazole inhibition, as was expected for these azole-resistant isolates which overexpress ATP-binding cassette transporter genes CDR1 and CDR2 . In addition to 3,6-diol, substantial amounts of obtusifolione were found after exposure to itraconazole . This toxic 3-ketosteroid was demonstrated previously to accumulate after itraconazole treatment in Cryptococcus neoformans and Histoplasma capsulatum but has not been reported in Candida isolates . Accumulation of obtusifolione correlated with nearly complete growth inhibition in these azole-resistant strains compared to that found in the susceptible parent strain, although the onset of growth inhibition only occurred at higher concentrations of itraconazole . ERG25 and ERG26 are the only genes assigned to the 4-demethylation process, of which the 3-ketoreductase is part . To verify whether mutations in these ERG25 genes contributed to obtusifolione accumulation, their nucleotide sequences were determined in all three related isolates . No mutations in ERG25 alleles of isolates C48 and C56 were found, suggesting that this gene is not involved in obtusifolione accumulation . The molecular basis for the accumulation of this sterol in these two strains remains to be established.

Allergy, 1999 Oct, 54(10), 1067 - 73
Cross-reacting IgE and IgG antibodies to Pityrosporum ovale mannan and other yeasts in atopic dermatitis; Lintu P et al.; Atopic dermatitis (AD) patients often demonstrate positive skin prick test results and serum IgE antibodies to a range of different yeasts . This has been thought to be due to cross-reactivity . In this study, the cross-reactivity of IgE and IgG antibodies between mannan and crude antigens of Pityrosporum ovale, Candida albicans, and Saccharomyces cerevisiae and crude antigens of Cryptococcus albidus and Rhodotorula rubra was examined by RAST and ELISA inhibition with two serum pools of AD patients . We found cross-reacting IgE and IgG antibodies . In the IgE response, the main cross-reacting pattern was the mannan region, although inhibition could be achieved also with crude antigens of C . albicans, S . cerevisiae, and, to some extent, C . albidus . P . ovale was the most potent inhibitor of IgE-binding components, and against it the highest IgE antibody levels were detected in AD serum pools . In contrast, C . albicans was found to be the most important inducer of IgG antibodies, since the IgG level against P . ovale mannan in both AD serum pools was very low . Cross-reacting antibodies were also seen in ELISA inhibition with both crude and mannan antigens, but since the IgG antibody level of P . ovale mannan in AD serum pools was low, further studies are needed to confirm the IgG results.

Postgrad Med J, 1999 May, 75(883), 297 - 8
Massive pleural effusions in cryptococcal meningitis; Wong CM et al.; Cryptococcal infection uncommonly presents with pulmonary manifestations and even more rarely so as massive bilateral effusions . Pleural involvement is usually associated with underlying pulmonary parenchymal lesions and is unusual while on antifungal therapy . We report a patient with cryptococcal meningitis who, while on intravenous 5-flucytosine and amphotericin B, developed life-threatening bilateral massive pleural effusions with evidence of spontaneous resolution, consistent with prior hypothesis of antigenic stimulation as the cause of pleural involvement.

Infect Immun, 1999 Nov, 67(11), 6139 - 44
Isolation, characterization, cDNA cloning, and antimicrobial properties of two distinct subfamilies of alpha-defensins from rhesus macaque leukocytes; Tang YQ et al.; Experiments to isolate and characterize rhesus macaque myeloid alpha-defensins (RMADs) were conducted . Seven RMAD peptides were isolated and sequenced, and the cDNAs encoding six of these peptides and one other alpha-defensin from bone marrow were also characterized . Four of the RMADs were found to be highly similar to human neutrophil alpha-defensins HNP-1 to HNP-3, while the remaining four peptides were much more similar to human enteric alpha-defensin HD-5 . Two alpha-defensin pairs differed only by the presence or absence of an additional arginine at the amino termini of their mature peptides, indicative of alternate posttranslational processing . The primary translation products of RMAD-1 to -8 are 94- and 96-amino-acid prepropeptides that are highly similar to those of human alpha-defensins . Immunolocalization experiments revealed a granular cytoplasmic pattern in the cytoplasms of neutrophils, indistinguishable from the pattern observed after immunostaining of human myeloid alpha-defensins in polymorphonuclear leukocytes . Each of the purified peptides was tested for its in vitro activities against Staphylococcus aureus 502a, Listeria monocytogenes EGD, Escherichia coli ML35, and Cryptococcus neoformans 271A . Several of the peptides were microbicidal for the gram-positive bacteria and C . neoformans at defensin concentrations in the range of 2 to 5 microM . All of the peptides were bacteriostatic against E . coli, but none were bactericidal for this organism . This study is the first to characterize the sequences and activities of alpha-defensins from nonhuman primates, data that should aid in delineating the role of these peptides in rhesus macaque host defense.

Infect Immun, 1999 Nov, 67(11), 6076 - 83
Phenotypic switching in Cryptococcus neoformans results in changes in cellular morphology and glucuronoxylomannan structure; Fries BC et al.; Cryptococcus neoformans strains exhibit variability in their capsular polysaccharide, cell morphology, karyotype, and virulence, but the relationship between these variables is poorly understood . A hypovirulent C . neoformans 24067A isolate, which usually produces smooth (SM) colony types, was found to undergo phenotypic switching and to produce wrinkled (WR) and pseudohyphal (PH) colony types at frequencies of approximately 10(-4) to 10(-5) when plated on Sabouraud agar . Cells from these colony types had large polysaccharide capsules and PH morphology, respectively . Scanning electron microscopy showed that different colony types were the result of altered cellular packing in the colony . Phenotypic switching was associated with quantitative and qualitative changes in capsular polysaccharide . Specifically, the glucuronoxylomannan (GXM) of the WR polysaccharide differed in the proportion of structural reporter groups and in increased xylose residue content linked at the 4 to 0 position . The relative virulence of the colony types was WR > PH > SM, as measured by CFU in rat lungs after intratracheal infection . Karyotype instability was observed in strain 24067A and involved primarily two chromosomes . Colonies with an alternative colony type exhibited more karyotype changes, which did not revert to the original karyotype in reverted colonies . In summary, this study revealed that phenotypic switching in C . neoformans (i) can produce WR colonies consisting of cells with either large capsule or PH morphology, (ii) is associated with production of structurally different GXM, (iii) is commonly associated with karyotype changes, (iv) can produce cells of PH morphology, and (v) can increase the virulence of a strain . Hence, phenotypic switching is an adaptive mechanism linked to virulence that can generate cell types with very different biological characteristics.

Infect Immun, 1999 Nov, 67(11), 6034 - 9
Laccase protects Cryptococcus neoformans from antifungal activity of alveolar macrophages; Liu L et al.; While laccase of Cryptococcus neoformans is implicated in the virulence of the organism, our recent studies showing absence of melanin in the infected mouse brain has led us to a search for alternative roles for laccase in cryptococcosis . We investigated the role of laccase in protection of C . neoformans against murine alveolar macrophage (AM)-mediated antifungal activity by using a pair of congenic laccase-positive (2E-TUC) and laccase-deficient (2E-TU) strains . The laccase-positive cells with laccase derepression were more resistant to the antifungal activity of AM than a laccase-deficient strain ({28.9 +/- 1.2}% versus {40.2 +/- 2.6}% killing) . Addition of L-dopa to Cryptococcus to produce melanin in a laccase-positive strain resulted in a slight increase in protection of C . neoformans from the antifungal activity of macrophages ({25.4 +/- 3.4}% versus {28.9 +/- 1.2}% killing) . Recombinant cryptococcal laccase exhibited iron oxidase activity in converting Fe(II) to Fe(III) . Moreover, recombinant laccase inhibited killing of C . neoformans by hydroxyl radicals catalyzed by iron in a cell-free system . Addition of the hydroxyl radical scavenger mannitol or dimethyl sulfoxide to AMs prior to the introduction of cryptococcal cells decreased killing of both strains and reduced the difference in susceptibility between the laccase-positive and laccase-deficient strains . Furthermore, laccase-mediated protection from AM killing was inhibited by the addition of Fe(II), presumably by overcoming the effects of the iron oxidase activity of cryptococcal laccase . These results suggest that the iron oxidase activity of laccase may protect C . neoformans from macrophages by oxidation of phagosomal iron to Fe(III) with a resultant decrease in hydroxyl radical formation.

Bioorg Med Chem, 1999 Sep, 7(9), 1933 - 40
Synthesis and antifungal activity of coumarins and angular furanocoumarins; Sardari S et al.; Angelicin, a naturally occurring furanocoumarin, that showed antifungal activity, was considered as a lead structure for a group of synthetic coumarins . Antifungal activities of the synthesized coumarins and angelicin derivatives were reported against Candida albicans, Cryptococcus neoformans, Saccharomyces cerevisiae and Aspergillus niger . Human cell line cytotoxicity of several coumarins was evaluated against KB cells . Angelicin and several potent antifungals showed to be non-toxic in this assay.

Zentralbl Veterinarmed B, 1999 Sep, 46(7), 443 - 51
Isolates of fungi from symptomatic carthorses in Awassa, Ethiopia; Etana D; Samples were collected from clinically infected carthorses in Awassa . Fungus species affecting the carthorses were identified . Eight genera of fungal groups were isolated from swabs and skin scrapes taken from symptomatic horses . These included Aspergillus spp., Histoplasma spp., Penicillium spp., Microsporum spp., Trichophyton spp., yeast cells of Candida spp., Cryptococcus spp . and Geotrichum spp . The most frequent isolates were from the genera Aspergillus (48%), Penicillium (39.2%) and Trichophyton (31.6%) . Clinical findings are reported, the economic, zoonotic and pathogenetic importance of fungi causing dermatomycoses is discussed, and further studies are recommended.

J Immunol, 1999 Nov 1, 163(9), 4642 - 6
Cutting edge: Role of C-C chemokine receptor 5 in organ-specific and innate immunity to Cryptococcus neoformans; Huffnagle GB et al.; After intratracheal inoculation of the AIDS-associated pathogen Cryptococcus neoformans, 12-wk survival was >90% for CCR5+/+ mice but <25% for CCR5-/- mice . There were no defects in lung leukocyte recruitment (wk 5), pulmonary clearance, or delayed-type hypersensitivity in CCR5-/- mice . However, CCR5-/- mice had defects in leukocyte recruitment into the brain and, strikingly, in elimination of cryptococcal polysaccharide from the brain . In nonimmune CCR5-/- mice, there was a significant defect in macrophage recruitment after challenge with shed cryptococcal products (C . neoformans filtrate Ag) but not other nonspecific stimuli . Thus, CCR5 plays specific roles in innate immunity and organ-specific leukocyte trafficking during host defense against C . neoformans.

Int J Tuberc Lung Dis, 1999 Oct, 3(10), 908 - 12
Bone marrow cultures for the diagnosis of mycobacterial and fungal infections in patients infected with the human immunodeficiency virus; Talbot EA et al.; SETTING: University medical center . OBJECTIVE: To determine the value of bone marrow cultures for mycobacteria and fungi in patients infected with the human immunodeficiency virus (HIV) . DESIGN: Retrospective review of charts and laboratory records . RESULTS: From 1992-1996, 1225 bone marrow specimens were submitted for mycobacterial and fungal cultures . The number of specimens submitted,declined sharply from 435 in 1992 to 94 in 1996 (P = 0.002 for trend) . The yield remained stable . Thirty-one of 1225 specimens grew mycobacteria or fungi; 26 isolates were from 24 HIV-infected patients . These 24 patients were infected with Mycobacterium avium complex (19), M . tuberculosis (one), M . chelonae (one), Histoplasma capsulatum (two), and Cryptococcus neoformans (one) . All 24 HIV-infected patients had a culture submitted from at least one other site within 4 weeks of the positive bone marrow culture . The identical organism was grown from another site (usually blood) in 18 of these 24 patients . The bone marrow culture provided the only positive result in six patients and the first positive result in eight patients . CONCLUSIONS: Utilization of bone marrow cultures for mycobacteria and fungi declined at our institution . Bone marrow and blood cultures were highly concordant . However, the majority of positive bone marrow cultures provided useful information.

Tidsskr Nor Laegeforen, 1999 Sep 10, 119(21), 3132 - 4
{Cryptococcal meningitis in a patient without known predisposing disease}; Christensen A et al.; Cryptococcal meningitis is a rare disease . It may occur as a superinfection in AIDS patients or other immunosuppressed patients . We describe a case of cryptococcal meningitis in a non-immunosuppressed patient . Initial symptoms were fatigue, depression and headache . A correct diagnosis was made after two weeks based on microscopic examination of cerebrospinal fluid . The patient died after six days on antimycotic therapy . Cryptococcosis is a difficult diagnosis, as our case illustrates . Psychiatric symptoms are often the first clinical manifestations . Early diagnosis is crucial for the outcome . A short overview on cryptococcosis is given.

Med Mycol, 1999 Oct, 37(5), 367 - 9
Feral pigeons as carriers of Cryptococcus laurentii, Cryptococcus uniguttulatus and Debaryomyces hansenii; Mattsson R et al.; We collected fresh droppings and cloaca samples from feral pigeons Columba livia in the southern Swedish city of Malmo, and isolated the following fungi: Debaryomyces hansenii var . hansenii, Cryptococcus laurentii and Cryptococcus uniguttulatus . The first two species are known to be pathogenic to humans . No strains of Cryptococcus neoformans var . neoformans were found . Our results indicate that feral pigeons can be carriers of medically significant fungi other than Cryptococcus neoformans var . neoformans.

Med Mycol, 1999 Oct, 37(5), 339 - 44
Antifungal activity of cerebrospinal fluid against Cryptococcus neoformans and Candida species; Brummer E et al.; The effect of human cerebrospinal fluid (CSF) on the growth of Cryptococcus neoformans and Candida species was tested in RPMI-1640 . CSF alone was highly fungistatic for both yeasts and inhibited growth in a concentration-dependent manner . Unlike human serum, CSF did not collaborate with fluconazole for killing C . neoformans . Molecular sieve fractionation of CSF on a G-200 Sephadex column yielded a highly antifungal fraction with a molecular weight around 66 kDa . On SDS-PAGE this fraction migrated as a major and a minor band corresponding to the mobility of bovine serum albumin . These novel findings suggest that CSF contains a factor(s) that provides resistance to the growth of C . neoformans or Candida species.

Med Mycol, 1999 Oct, 37(5), 331 - 8
Asymptomatic carriage of Cryptococcus neoformans in the nasal cavity of the koala (Phascolarctos cinereus); Connolly JH et al.; Over a 22-month period, sequential nasal and skin swabs were obtained from 52 healthy captive koalas (Phascolarctos cinereus) from the Sydney region . Cryptococcus neoformans was isolated in 17 koalas from 64 of 262 (24%) nasal swabs and from nine of 262 (3%) skin swabs . Prevalence of nasal colonization varied seasonally from 12% (3/25) to 38% (10/26) . Cryptococcus neoformans var . gattii alone was cultured from 37, var . neoformans alone from 22 and both varieties from five nasal swabs . Of 33 koalas sampled on three or more occasions, organisms were isolated persistently from six, occasionally from eight and never from 19 . Two koalas were persistently and heavily (>/=100 colonies/plate) colonized by C . neoformans var . gattii and two with var . neoformans . Isolation of C . neoformans var . gattii from the skin was low grade and sporadic . No koalas from which C . neoformans was persistently isolated showed clinical signs of cryptococcosis and all except one had a negative latex cryptococcal antigen test, therefore the nasal cavity was presumed to be colonized by, rather than infected with, C . neoformans . Preliminary observations of koalas from Coffs Harbour indicated a much higher prevalence of colonization by C . neoformans, suggesting that environmental factors influenced the extent of carriage by C . neoformans.

Eur J Clin Microbiol Infect Dis, 1999 Aug, 18(8), 587 - 90
Disseminated infection due to Nocardia transvalensis coincident with Cryptococcus neoformans variety gattii meningitis; Dyer JR et al.; A case of meningitis due to Cryptococcus neoformans var . gattii coincident with disseminated Nocardia transvalensis infection is reported . Nocardia infection initially progressed despite high-dose antimicrobial therapy . Although a specific immunologic defect could not be defined, in vitro lymphocyte proliferation in response to stimulation with the Nocardia isolate was reduced . It is proposed that coinfection with Cryptococcus neoformans may have contributed to the observed impairment of lymphocyte function, leading to disseminated Nocardia disease and a suboptimal treatment response.

J Infect Dis, 1999 Nov, 180(5), 1637 - 47
Enhanced sensitivity of tumor necrosis factor/lymphotoxin-alpha-deficient mice to Cryptococcus neoformans infection despite increased levels of nitrite/nitrate, interferon-gamma, and interleukin-12; Rayhane N et al.; The cytokine network and infection severity were characterized during disseminated cryptococcosis in tumor necrosis factor (TNF)- and lymphotoxin (Lt)-alpha-deficient mice . On day 16, the fungus burden was higher and median survival time was reduced, as was polymorphonuclear leukocyte infiltrate in the brains of knockout mice . TNF/Lt-alpha-deficient mice had lower levels of interleukin (IL)-6 in lungs and brains, IL-1beta, and the chemokine KC in brain and spleen and of the chemokine monocyte chemoattractant protein (MCP)-1 in spleen than control animals . In contrast, higher levels of IL-6, IL-10, and MCP-1 in plasma and higher levels of IL-12, interferon (IFN)-gamma, and nitrite/nitrate were found in all compartments of TNF/Lt-alpha-deficient mice . These data confirm that TNF or Lt-alpha is a key cytokine for the anticryptococcal response and demonstrate its major role for the induction of IL-1beta, IL-6, and KC in the brain; however, its presence is not a prerequisite for IL-12, IFN-gamma, and nitrite/nitrate production.

J Infect Dis, 1999 Nov, 180(5), 1542 - 9
Neutrophils from patients with advanced human immunodeficiency virus infection have impaired complement receptor function and preserved Fcgamma receptor function; Monari C et al.; Interleukin (IL)-8 production by human polymorphonuclear leukocytes (PMNL) to Cryptococcus neoformans is related to complement activation . Generation of the bioactive fragments C3a and C5a is responsible for IL-8 release . IL-8 production was analyzed in response to C . neoformans by PMNL from persons with early- and late-stage (>400 and <200 CD4 cells/mm3, respectively) human immunodeficiency virus (HIV) infection who were at high risk for cryptococcosis . IL-8 release by PMNL from persons with early-stage infection and from healthy donors was similar; however, PMNL from persons with late-stage HIV infection had significantly impaired IL-8 production, which correlated with reduced IL-8 response to C3a and C5a proteins and decreased CD88 expression . Addition of murine monoclonal antibody (MAb) 18B7 promoted phagocytosis and restored IL-8 release consistent with integrity of FcgammaRIII . These results provide evidence for a selective defect in CD88 expression on PMNL from persons with late-stage HIV infection . However, Fcgamma receptor expression in PMNL appears to be intact and allows MAb to glucuronoxylomannan to positively influence PMNL function.

J Infect Dis, 1999 Nov, 180(5), 1526 - 35
Quantitative and qualitative differences in the serum antibody profiles of human immunodeficiency virus-infected persons with and without Cryptococcus neoformans meningitis; Fleuridor R et al.; The importance of humoral immunity for resistance to Cryptococcus neoformans is uncertain . A case-controlled study of the human antibody response to C . neoformans comparing the serum antibody profiles of human immunodeficiency virus (HIV)-infected persons who did (HIV+/CM+) or did not (HIV-infected controls) develop cryptococcal meningitis (CM) and HIV-uninfected persons with samples obtained from the Multicenter AIDS Cohort Study was performed . Total immunoglobulin concentrations were determined, and the specificity, isotype, and idiotype expression of antibodies to C . neoformans capsular glucuronoxylomannan were analyzed by ELISA . Compared with the HIV+/CM+ group, the HIV-infected control group had significantly lower levels of total IgM, IgA, and antibodies expressing a certain VH3 determinant . The HIV-infected control group manifested an increase in immunoglobulin levels with a decrease in CD4 lymphocytes . The findings suggest a possible association between reduced expression of certain immunoglobulin subsets and HIV-associated CM.

AIDS, 1999 Oct 1, 13(14), 1963 - 9
Clinical disease associated with HIV-1 subtype B' and E infection among 2104 patients in Thailand; Amornkul PN et al.; BACKGROUND: Two HIV-1 envelope subtypes have accounted for virtually all infections in Thailand: subtype B' (Thai B), found mainly in injection drug users (IDU), and subtype E, found in over 90% of sexually infected persons and an increasing proportion of IDU in recent years . It remains unclear whether there are differences in pathogenesis associated with these HIV-1 subtypes . METHODS: From November 1993 to June 1996, demographic, risk, clinical, and laboratory data were collected by enhanced surveillance from HIV-infected inpatients (> or =14 years) at an infectious disease hospital near Bangkok . HIV-1 subtype was determined by V3-loop peptide enzyme immunoassay (EIA) . Because of confounding, multivariate analyses were stratified by risk category and controlled for sex and age . RESULTS: The infecting HIV-1 subtype was determined for 2104 (94.9%) of 2217 HIV-infected patients with complete data: 284 (13.5%) were subtype B', 1820 (86.5%) were E . Specimens from 113 (5.1%) patients were non-reactive or dually reactive on peptide EIA and were excluded . Among IDU, 199 (67.2%) had subtype B', and 97 (32.7%) had E . IDU accounted for 70.1% (199/284) of patients with subtype B' and 5.3% (97/1820) of those with E . Patients infected with HIV-1 subtypes B' or E had similar spectrums of opportunistic infections (OI), levels of immunosuppression, and in-hospital mortality rates . Of patients who did not inject drugs, more patients infected with subtype E had extrapulmonary cryptococcosis than those with subtype B' (adjusted odds ratio, 2.6; 95% confidence interval, 1.28-5.37) . CONCLUSION: HIV-1 subtypes B' and E seem to be associated with similar degrees of immunosuppression and, with one exception, with similar OI patterns . These data do not suggest an association between HIV-1 subtype and differences in pathogenicity.

Fungal Genet Biol, 1999 Oct, 28(1), 1 - 5
On the origins of congenic MATalpha and MATa strains of the pathogenic yeast Cryptococcus neoformans; Heitman J et al.; The basidiomycetous yeast Cryptococcus neoformans infects humans and causes a meningoencephalitis that is uniformly fatal if untreated . The organism has a defined sexual cycle involving mating of haploid MATa and MATalpha strains, gene disruption by transformation and homologous recombination is now readily accomplished, and robust animal models for infection have been well established . In addition, a pair of congenic MATalpha and MATa haploid strains have been constructed that permit detailed studies on physiology and virulence by classical genetic approaches . These strains represent a valuable resource for further studies in this organism, and the genomic sequence of one of these strains, JEC21 (=B-4500), was recently chosen to be sequenced by an international consortium . Because of the importance of these strains for genetic studies in C . neoformans and the fact that the genomic sequence of one of these strains is in progress, we review here how these congenic strains were originally constructed .

Neth J Med, 1999 Sep, 55(3), 118 - 27
Therapy of invasive fungal infections; Kullberg BJ et al.; During the past decades, the incidence and severity of invasive fungal infections has been increasing . These events have lead to the development of new antifungal agents, and amphotericin B no longer is the standard therapy for a variety of invasive mycoses . Fluconazole has become the drug of choice for treatment of C . albicans infections in non-neutropenic as well as neutropenic patients . For treatment of cryptococcal meningitis, amphotericin B with flucytosine, followed by fluconazole consolidation therapy is used . For therapy of invasive aspergillosis, the lipid formulations of amphotericin B may be associated with reduced toxicity and allow for larger doses, although the efficacy of each of the formulations still has to be established in randomized trials . Finally, treatment modalities aimed at improving host defense mechanisms, including adjunctive therapy with rG-CSF for disseminated candidiasis and rG-CSF-elicited granulocyte transfusions, are under way.

An Med Interna, 1999 Aug, 16(8), 417 - 9
{Lumbosacral polyradiculomyelitis caused by herpes simplex virus (HSV) in a patient with AIDS}; Miguelez M et al.; To our knowledge this is the first description of lumbosacral polyradiculopathy produced by herpes simplex virus (HSV) without coinfection by cytomegalovirus (CMV) in a patient with HIV infection . The acute lumbosacral polyradiculomyelitis (ALP) in patients with AIDS is a well defined nosologic entity and classically associated to CMV infection . However, this pathology can be due to others etiologies as toxoplasmosis, syphilis, lymphoma, tuberculosis, cryptococcus, varicella-zoster virus fVZV), Epstein-Barr virus (EBT) and HSV associated CMV . The case report was documented with findings in cerebrospinal fluid, magnetic resonance imaging and was confirmed by detection of HSV DNA by polymerase chain reaction . CMV DNA was not detected and no clinical features of CMV disease was seen . Therapy with foscarnet was successful . This drug without gancyclovir associated has rarely been employed in the treatment of ALP . We believe that foscarnet may be a valuable alternative therapy for cases of ALP of suspected acyclovir-resistant herpes virus infection.

South Med J, 1999 Sep, 92(9), 927 - 9
Rapid respiratory deterioration and sudden death due to disseminated cryptococcosis in a patient with the acquired immunodeficiency syndrome; Penmetsa S et al.; We report the case of a patient with the acquired immunodeficiency syndrome (AIDS) whose death occurred within 30 hours of hospitalization due to disseminated cryptococcosis, manifested by dizziness, cough, and shortness of breath . The clinical picture was consistent with pneumocystis pneumonia, and antibiotic therapy with corticosteroids was initiated . Despite initial improvement, the patient's condition quickly worsened, resulting in cardiorespiratory arrest and death . Autopsy revealed cryptococci in several organs . Sudden, rapid deterioration and death are rare consequences of disseminated cryptococcosis, and steroids may worsen the course of the disease . On the basis of this case and review of similar cases in the literature, we recommend early consideration of disseminated cryptococcosis in AIDS patients with pneumonia . Early diagnosis and appropriate therapy are essential to reduce morbidity and mortality.

FEMS Immunol Med Microbiol, 1999 Sep, 25(4), 391 - 402
Chemokine responses and accumulation of inflammatory cells in the lungs of mice infected with highly virulent Cryptococcus neoformans: effects of interleukin-12; Kawakami K et al.; We examined the mechanisms involved in the development of lung lesions after infection with Cryptococcus neoformans by comparing the histopathological findings and chemokine responses in the lungs of mice infected with C . neoformans and assessed the effect of interleukin (IL) 12 which protects mice from lethal infection . In mice infected intratracheally with a highly virulent strain of C . neoformans, the yeast cells multiplied quickly in the alveolar spaces but only a poor cellular inflammatory response was observed throughout the course of infection . Very little or no production of chemokines, including MCP-1, RANTES, MIP-1alpha, MIP-1beta and IP-10, was detected at the mRNA level using RT-PCR as well as at a protein level in MCP-1, RANTES and MIP-1alpha . In contrast, intraperitoneal administration of IL-12 induced the synthesis of these chemokines and a marked cellular inflammatory response involving histiocytes and lymphocytes in infected mice . Our findings were confirmed by flow cytometry of intraparenchymal leukocytes obtained from lung homogenates which showed IL-12-induced accumulation of inflammatory cells consisting mostly of macrophages and CD4+ alphabeta T cells . On the other hand, C-X-C chemokines including MIP-2 and KC, which attract neutrophils, were produced in infected and PBS-treated mice but treatment with IL-12 showed a marginal effect on their level, and neutrophil accumulation was similar in PBS- and IL-12-treated mice infected with C . neoforman . Our results demonstrate a close correlation between chemokine levels and development of lung lesions, and suggest that the induction of chemokine synthesis may be one of the mechanisms of IL-12-induced protection against cryptococcal infection.

Infect Immun, 1999 Oct, 67(10), 5477 - 9
Evidence that Cryptococcus neoformans is melanized in pigeon excreta: implications for pathogenesis; Nosanchuk JD et al.; Structures similar to the melanin "ghosts" of melanized cryptococcal cells were isolated from pigeon excreta contaminated with Cryptococcus neoformans, and their growth in pigeon excreta supported melanization . The results suggest that environmental C . neoformans cells are melanized and imply that initial infection may involve exposure to melanized cells.

Infect Immun, 1999 Oct, 67(10), 4994 - 5000
An opsonizing monoclonal antibody that recognizes a noncapsular epitope expressed on Cryptococcus neoformans; Merkel GJ et al.; A mouse hybridoma secreting a monoclonal antibody (MAb) that bound a noncapsular epitope expressed on C . neoformans was developed by immunizing BALB/c mice with formalin-killed serotype A yeasts . The hybridoma, designated CSFi, secreted an immunoglobulin G2b MAb that reacted with all C . neoformans serotypes tested, including the acapsular mutant ATCC 52817 (Cap67) . Postsectioned immune electron microscopy revealed extensive binding of the MAb to the cell walls of both encapsulated and acapsular yeasts . Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blot analysis of secreted antigens recovered from concentrated culture supernatants from both encapsulated and acapsular strains was conducted . The results showed that this MAb bound predominantly to antigens with molecular masses of approximately 75 and 100 kDa . A competitive enzyme-linked immunosorbent assay was used to demonstrate that the MAb was not cross-reactive with purified glucuronoxylomannan derived from either serotypes A or D . Experiments conducted with mouse peritoneal phagocytes and the mouse phagocyte-like cell line, J774A.1, demonstrated that the CSFi MAb opsonized the yeasts and increased their adherence to both types of phagocytic cells . We conclude, therefore, that antibodies directed at noncapsular epitopes can serve as opsonins and may have a role in modulating cryptococcal infection.

J Microbiol Immunol Infect, 1998 Dec, 31(4), 233 - 9
Efficacy and adverse effects of higher dose amphotericin B monotherapy for cryptococcal meningitis in patients with advanced HIV infection; Hsieh SM et al.; Treatment with a low daily dose of amphotericin B (0.4 mg/kg) in AIDS patients with cryptococcal meningitis has been associated with low efficacy and high mortality . We report our successful clinical experiences on a higher daily dose of amphotericin B (0.8-1.0 mg/kg) monotherapy in treating cryptococcal meningitis from June 1994 to August 1997 in 13 cases of advanced HIV infection . Most of them (12/13) had at least one of several poor prognostic factors . The mean duration of amphotericin B administration was 26 days (range, 3 to 58 days) . Both microbiologically and clinically successful rates of treatment at the end of amphotericin B therapy were high (85%, 11/13) . The median duration of negative CSF culture post therapy was 17 days (range, 8 to 33 days) . Bone marrow toxicities were; thrombocytopenia (46%) and significant anemia (92%) after a mean of 9 days of treatment . Both, impaired renal function and hypokalemia, were seen in 10 cases (77%), while elevation of amylase and lipase values were present in 6 cases (46%) . Our report reveals that a higher daily dose of amphotericin B can achieve a high efficacy in treatment of cryptococcal meningitis in AIDS patients, even though most cases had poor prognostic factors and were in severe immunocompromised states . However, clinicians should monitor higher dose-related adverse effects carefully.

Nihon Kokyuki Gakkai Zasshi, 1999 Aug, 37(8), 667 - 71
{Primary pulmonary cryptococcosis confused with lung cancer by transbronchial biopsy: demonstration by three-dimensional computed tomography}; Terasaki H et al.; Routine chest radiographs of a 52-year-old woman displayed abnormal opacities in the right lower lung field . A computed tomographic (CT) scan of the chest disclosed irregular nodular opacities in the peripheral zone of the right S6 . Reconstructed three-dimensional images obtained by helical CT displayed the lesions as agglutinated small nodules, with other small nodules near the bronchi appearing as "tree-in-bud" formations . Specimens obtained by transbronchial lung biopsy of the right B6b revealed atypical glandular structures, thus leading to suspected adenocarcinoma of primary lung cancer . A right lower lobectomy was performed; cut-surface specimens of the resected tissues showed multiple lobulate, milky-white nodules near the bronchi . Histopathologic examination found no malignant cells . PAS staining revealed numerous cryptococcal organisms, thus yielding the final diagnosis of primary pulmonary cryptococcosis.

Nihon Kokyuki Gakkai Zasshi, 1999 Aug, 37(8), 614 - 8
{Primary pulmonary cryptococcosis diagnosed by medical examinations in 12 patients}; Nishida Y et al.; Primary pulmonary cryptococcosis is thought to be relatively less common than other lung mycoses, but recently there has been an increase in reports of patients with this disease . Our report covers 12 cases of primary pulmonary cryptococcosis in which the diagnosis was based on medical examinations . The patients consisted of 11 men and 1 woman, aged 27 to 58 years . Only 3 exhibited subjective symptoms . Roentgenograms showed cavitating tumor shadows in the lungs of 2 of the patients with subjective symptoms, and nodular shadows with diameters of 1.0 to 2.8 cm in all patients without subjective symptoms, indicating the possibility of lung cancer . The disease was diagnosed in 3 patients on the basis of transbronchial biopsy findings, and in 9 on the basis of needle aspiration biopsy findings . One patient was diabetic, but the others did not exhibit malignancies or other immunocompromised states . Antifungal drugs significantly reduced or eliminated the nodular shadows in 10 patients . Because patients with primary pulmonary cryptococcosis frequently lack subjective symptoms, prompt diagnosis is critical, particularly in view of the need to distinguish the disease from lung cancer.

East Afr Med J, 1998 Sep, 75(9), 516 - 9
Study of the aetiologic agents of meningitis in Kumasi, Ghana, with special reference to Cryptococcal neoformans; Frimpong EH et al.; OBJECTIVE: To evaluate the importance of Cryptococcus neoformans, an opportunistic in meningitis, in healthy and HIV infected patients in Kumasi, Ghana . DESIGN: A prospective study; isolating the aetiologic agents of meningitis from cerebrospinal fluid (CSF) using standard methods . SETTING: Kumasi city, Ashanti region of central Ghana . SUBJECTS: One thousand five hundred and seventy patients suspected of meningitis, including 28 HIV infected and AIDS patients . MAIN OUTCOME MEASURE: The pattern and distribution of the main etiologic agents of meningitis in Kumasi, Ghana shown . RESULTS: Of the 1570 CSF samples examined, 1256 (80%) showed no abnormality . Of the 314 (20%) with abnormalities, 147 were bacterial, and 167 diagnosed aseptic . No cryptococcal cells were found . CONCLUSION: There is a paucity of cryptococcal meningitis in Kumasi, Ghana, and it is not recommended to screen routinely for the fungus in meningitis investigations.

Postgrad Med, 1999 Jul, Spec No, 46 - 52
Oral itraconazole therapy for superficial, subcutaneous, and systemic infections . A panoramic view; De Rosso JQ et al.; The availability of the oral triazole agents itraconazole and fluconazole has revolutionized antifungal therapy . Although there are still some limitations and treatment failures, these agents have allowed for improved efficacy, increased safety, reduced morbidity, decreased mortality from systemic fungal disease, and a shift toward increased outpatient therapy for fungal infections that are not life-threatening . The treatment of superficial infections also has been enhanced by the development of effective intermittent and short-course regimens . Itraconazole exhibits broad-spectrum in vitro activity against several fungal organisms, including Trichophyton species, Candida albicans, Pityrosporum species, Aspergillus flavus, Aspergillus fumigatus, Blastomyces dermatitidis, Histoplasma capsulatum, Histoplasma capsulatum var duboisii, Sporothrix schenckii, and Cryptococcus neoformans . Animal model studies have confirmed the broad-spectrum in vivo activity of itraconazole . Multiple clinical studies and extensive clinical experience have substantiated the versatility of itraconazole, with good efficacy demonstrated in a wide variety of infections in humans . Itraconazole is approved for use in several countries for dermatomycoses, onychomycosis, oral-esophageal candidiasis, vaginal candidiasis, histoplasmosis, blastomycosis, aspergillosis, and fungal keratitis . Pulse therapy regimens for dermatomycoses and onychomycosis of the toenails or fingernails have been approved in several countries.

J Immunol, 1999 Oct 1, 163(7), 3907 - 13
Ig heavy chain complex-linked genes influence the immune response in a murine cryptococcal infection; Lovchik JA et al.; A murine pulmonary infection with Cryptococcus neoformans (Cne) has been used to determine mechanisms regulating effective T cell-mediated immunity in the lungs . In BALB/c and C.B-17 mice, following intratracheal deposition of Cne, the fungus initially grows rapidly and is then progressively cleared from the lungs . Cne clearance in C.B-17 mice requires CD4 and CD8 T cells, IFN-gamma, and NO . Clearance in congenic BALB/c mice proceeds more slowly than in C.B-17 mice, even though the only genetic difference between these strains is at the Ig H chain-containing region of chromosome 12 . Examination of the pulmonary immune response in the two strains revealed that both cleared lung Cne by T cell-dependent mechanisms and generated equivalent levels of NO . Furthermore, both strains recruited equal numbers of macrophages, lymphocytes, and neutrophils to the lungs, although BALB/c mice recruited higher numbers of eosinophils . Notably, leukocytes isolated from BALB/c lungs during infection secreted lower levels of IFN-gamma and higher levels of the Th2 cytokines IL-4 and IL-5 as compared with lung leukocytes from C.B-17 mice . Furthermore, serum levels of IgM, IgG1, IgG2a, and IgG3 anti-Cne Abs generated during infection were significantly greater in BALB/c mice than C.B-17 mice . These data suggest that although both BALB/c and C.B-17 mice clear pulmonary cryptococcosis through T cell-mediated mechanisms, Ig H chain-linked genes in BALB/c mice are associated with a decreased effectiveness of the host response, which we suggest might influence the balance in Th1/Th2 T cell subset development or increase anti-Cne Abs, or both.

Vet Pathol, 1999 Sep, 36(5), 445 - 8
Cryptococcus neoformans and Mycobacterium bovis causing granulomatous pneumonia in a goat; Gutierrez M et al.; An adult Blanca-Celtiberica doe originating from a goat herd with a high prevalence of tuberculosis was presented with respiratory signs . At necropsy, this goat had a diffuse and severe mycotic pneumonia associated with the presence of Cryptococcus neoformans concomitant with pulmonary focal caseous nodules from which Mycobacterium bovis was isolated . Microscopically, the mycotic lesion was a granulomatous pneumonia with many large foamy macrophages containing intracellular yeast bodies . The extensive mycotic changes, their granulomatous nature, and the lack of positive response to different immunologic tests for mycobacterial infection suggested an impaired immune status in this animal.

Curr Microbiol, 1999 Nov, 39(5), 302 - 5
Repeated ubiquitin genes in Trichophyton mentagrophytes; Kano R et al.; Using Trichophyton mentagrophytes cDNA as a template, we performed PCR amplification with the UB1S and UB1R primers for isolation of a partial fragment of T . mentagrophytes ubiquitin gene . We screened the T . mentagrophytes cDNA library prepared with Uni-ZAP phage vector by hybridization with the T . mentagrophytes ubiquitin fragment, UB1S-UB1R . The nucleotide and deduced amino acid sequences of T . mentagrophytes ubiquitin gene encoded two ubiquitin repeats in 162 bp to 624 bp . The deduced amino acid sequences of ubiquitin from T . mentagrophytes shared about 98% similarity with those of Saccharomyces cerevisiae, Neurospora crassa, and Cryptococcus neoformans.

J Clin Microbiol, 1999 Oct, 37(10), 3204 - 9
Epidemiological evidence for dormant Cryptococcus neoformans infection; Garcia-Hermoso D et al.; To date, the time of acquisition of a Cryptococcus neoformans infectious strain has never been studied . We selected a primer, (GACA)(4), and a probe, CNRE-1, that by randomly amplified polymorphic DNA (RAPD) analysis and restriction fragment length polymorphism (RFLP), respectively, regrouped strains from control samples of C . neoformans var . grubii environmental isolates according to their geographical origins . The two typing techniques were then used to analyze 103 isolates from 29 patients diagnosed with cryptococcosis in France . Nine of the 29 patients lived in Africa a median of 110 months prior to moving to France; 17 of the patients originated from Europe . Results showed a statistically significant clustering of isolate subtypes from patients originating from Africa compared to those from Europe . We conclude that the patients had acquired the C . neoformans infectious strain long before their clinical diagnoses were made.

Nervenarzt, 1999 Aug, 70(8), 732 - 7
{Cryptococcal meningoencephalitis in HIV infected immigrants . Diagnostic and therapeutic aspects with reference to evidence-based medicine"--an overview}; Weltermann BM et al.; Cryptococcal meningitis is one of the most frequent forms of meningoencephalitis in AIDS patients . Before the introduction of triazole antibiotics like difluconazole, the frequency of this meningitis among AIDS patients was 5-10%, yet declined during the last years . Clinically, nonspecific signs of a meningeal inflammation predominate while focal neurological signs are rare . Rapid institution of high-dose antimycotic therapy is the keystone to improve prognosis and decrease mortality . Antimycotics of choice are amphotericin B, fluconazole, and flucytosin . Diagnosis is established by detection of cryptococcal antigen in cerebrospinal fluid, microscopic demonstration of cryptococci using India-ink, and cryptococcal cultures . Recent developments aim to improve antimycotic therapies . During the last years, lifelong secondary prevention with difluconazole was established . Cryptococci which are resistant against fluconazole and amphotericin B are a special challenge, yet fortunately are rare . Longterm outcome of patients is determined by the progression of the underlying immunosuppression . Therefore, combination of secondary prophylaxis with modern antiviral substances is important . Clinical decision analysis in patients with suspected cryptococcal meningitis is presented using methods from evidence-based medicine.

J Bacteriol, 1999 Sep, 181(18), 5636 - 43
Isolation, characterization, and localization of a capsule-associated gene, CAP10, of Cryptococcus neoformans; Chang YC et al.; Cryptococcus neoformans is a pathogenic fungus which most commonly affects the central nervous system and causes fatal meningoencephalitis primarily in patients with AIDS . This fungus produces a thick extracellular polysaccharide capsule which is well recognized as a virulence factor . Here, we describe the isolation and characterization of a novel gene, CAP10, which is required for capsule formation . Complementation of the acapsular cap10 mutant produced an encapsulated strain and the deletion of CAP10 from a wild strain resulted in an acapsular phenotype . The molecular mass of the hemagglutinin epitope-tagged Cap10p is about 73 kDa, which is similar to the size predicted from sequence analysis . When CAP10 was fused with a hybrid green fluorescent protein construct, the fluorescence signals appeared as patches in the cytoplasm . Using a reporter gene construct, we found that CAP10 was expressed at high levels in late-stationary-phase cells . In addition, we found that the expression levels of CAP10 are modulated by the transcriptional factor STE12alpha . Deletion of STE12alpha downregulated the expression levels of CAP10 while overexpression of STE12alpha upregulated the expression levels of CAP10 . Animal model studies indicate that deletion of the CAP10 gene results in the loss of virulence, and complementation of the acapsular phenotype of cap10 restores virulence . Thus, CAP10 is required for capsule formation and virulence.

J Immunol, 1999 Sep 15, 163(6), 3496 - 502
Cloning the antibody response in humans with inflammatory central nervous system disease: analysis of the expressed IgG repertoire in subacute sclerosing panencephalitis brain reveals disease-relevant antibodies that recognize specific measles virus antigens; Burgoon MP et al.; The presence of increased IgG in the brains of humans with infectious and inflammatory CNS diseases of unknown etiology such as multiple sclerosis may be a clue to the cause of disease . For example, the intrathecally synthesized oligoclonal bands (OGBs) in diseases such as subacute sclerosing panencephalitis (SSPE) or cryptococcal meningitis have been shown to represent Ab directed against the causative agents, measles virus (MV) or Cryptococcus neoformans, respectively . Using SSPE as a model system, we have developed a PCR-based strategy to analyze the repertoire of IgG V region sequences expressed in SSPE brain . We observed abnormal expression of germline V segments, overrepresentation of particular sequences that correspond to the oligoclonal bands, and substantial somatic mutation of most clones from the germline, which, taken together, constitute features of Ag-driven selection in the IgG response . Using the most abundant or most highly mutated gamma H chain and kappa or lambda L chain sequences in various combinations, we constructed functional Abs in IgG mammalian expression vectors . Three Abs specifically stained MV-infected cells . One Ab also stained cells transfected with the MV nucleoprotein, and a second Ab stained cells transfected with the MV-fusion protein . This technique demonstrates that functional Abs produced from putative disease-relevant IgG sequences can be used to recognize their corresponding Ags.

J Small Anim Pract, 1999 Aug, 40(8), 387 - 91
Intra-abdominal cryptococcosis in two dogs; Malik R et al.; Intra-abdominal cryptococcosis was diagnosed in two young dogs . The first, an entire male border collie, was presented with vomiting . An abdominal mass detected during physical examination proved to be cryptococcal mesenteric lymphadenitis on exploratory laparotomy . The second dog, a female neutered giant schnauzer, was presented with neurological signs suggestive of encephalopathy . Intestinal cryptococcal granulomas were detected in an extensive diagnostic investigation which included abdominal ultrasonography . The gastrointestinal tract was considered the most likely portal of entry for cryptococcal organisms in both cases . Both dogs were treated using surgery and multiagent antifungal chemotherapy . The first case succumbed despite therapy, while the second dog was treated successfully as gauged by return to clinical normality and a substantial decline in the cryptococcal antigen titre which continued to fall after cessation of treatment.

Postgrad Med J, 1999 Jul, 75(885), 387 - 90
HIV infection and seizures; Garg RK; New-onset seizures are frequent manifestations of central nervous system disorders in patients infected with human immunodeficiency virus (HIV) . Seizures are more common in advanced stages of the disease, although they may occur early in the course of illness . In the majority of patients, seizures are of the generalised type . Status epilepticus is also frequent . Associated metabolic abnormalities increase the risk for status epilepticus . Cerebral mass lesions, cryptococcal meningitis, and HIV-encephalopathy are common causes of seizures . Phenytoin is the most commonly prescribed anticonvulsant in this situation, although several patients may experience hypersensitivity reactions . The prognosis of seizure disorders in HIV-infected patients depends upon the underlying cause.

J Antimicrob Chemother, 1999 Aug, 44(2), 223 - 8
In-vitro activity of dicationic aromatic compounds and fluconazole against Cryptococcus neoformans and Candida spp; Del Poeta M et al.; We investigated the in-vitro activity of three selected dicationic aromatic compounds for nine clinical isolates of Cryptococcus neoformans and 93 clinical isolates of Candida spp., representing 12 different species, using a broth macrodilution method following NCCLS recommendations . All the clinical isolates were also tested for fluconazole susceptibility . The in-vitro data demonstrate that compounds 39 and 57 have excellent in-vitro activity for all tested strains (MIC 0.19-1.56 mg/L) except Candida pelliculosa . Moreover, compound 39 showed excellent in-vitro fungicidal activity against Candida krusei, Candida glabrata, Candida lusitaniae and Cryptococcus neoformans with MFCs in the range 0.39-6.25 mg/L . Both compounds 39 and 57 showed excellent in-vitro activity against fluconazole-resistant Candida albicans isolates, including a C . albicans strain that contains all known fluconazole-resistant mechanisms . Comparing MIC data from compounds 21, 39 and 57 with fluconazole, we found a statistically significant difference only with compound 39 (P = 0.043) . However, comparing MFC data from compounds 21, 39 and 57 with fluconazole, we found statistically significant differences with all three compounds (P < 0.00001) . These data indicate the potential antifungal breadth of two bis-benzimidazoles (compounds 39 and 57) as antifungal agents against yeasts . If it can be determined that compounds 39 and 57 are effective and non-toxic in vivo, the prospect of these compounds as clinically useful antifungal agents will be enhanced.

Immunology, 1999 Sep, 98(1), 27 - 35
B7 costimulatory ligand regulates development of the T-cell response to Cryptococcus neoformans; Monari C et al.; The contribution of B7 molecules to the induction and maintenance of the T-cell response to the human pathogenic fungus Cryptococcus neoformans was investigated . T-cell activation by C . neoformans was regulated by B7 molecules . This costimulatory signal was necessary for initiation and maintenance of the T-cell response, through early and late requirements for B7-CD28 interaction . Blocking B7-2 inhibited the normal T-cell proliferative response . This inhibition was due, in part, to a reduced capability of T cells to produce interleukin-2 (IL-2) . In contrast, the same T-cell population produced more interferon-gamma . Suppression of the normal lymphoproliferation and IL-2 secretion responses to encapsulated C . neoformans by antibodies to B7 was largely reversed by addition of the monoclonal antibody 2H1, that is reactive with the major capsular polysaccharide, glucuronoxylomannan . Overall, our data indicate that B7 molecules play a critical role in T-cell activation by C . neoformans and suggest that appropriate manipulation could drive T helper type 1 cell development.

J Infect, 1999 Jul, 39(1), 32 - 7
AIDS-associated cryptococcal meningitis in Rwanda (1983-1992): epidemiologic and diagnostic features; Bogaerts J et al.; OBJECTIVES: to document the trend of AIDS-associated Cryptococcus neoformans meningitis (CM) in Kigali, Rwanda, during 1983-1992, and to highlight some diagnostic and epidemiological features of the disease . METHODS: during the study period, 3476 cerebrospinal fluid (CSF) specimens from 2824 adults (1578 men, 1246 women) were analysed in the Laboratory of Microbiology at the Centre Hospitalier de Kigali, Rwanda, Central Africa, using direct examination, culture and detection of the cryptococcal antigen (CrAg) in the CSF . RESULTS: CM was diagnosed among 549 (19%) patients (347 men, 202 women) and was by far the leading cause of meningitis before Neisseria meningitidis (n=115), Streptococcus pneumoniae (n=68), Mycobacterium tuberculosis (n=26) . E . coli, Klebsiella pneumoniae, non-typhoid Salmonella (n=l5) and streptococci (n=4) . The number of CM increased from one case in 1983 to 130 new cases in 1992 . All 293 tested CM patients had HIV-1 antibodies . The male/female ratio declined from 3.31 during 1983-1987 to 1.58 during 1988-1992 . CM showed a seasonal fluctuation, the highest number of infections being observed during the long rainy season . The sensitivity and specificity of the latex test for diagnosing CM was 98% and 99%, respectively . Cryptococcus neoformans var . gattii was cultured from eight (1.6%) of the 499 culture positive patients . CONCLUSION: CM is an important opportunistic infection among AIDS patients in Central Africa . It remains a problematic diagnosis in areas with limited diagnostic facilities.

J Pharm Pharmacol, 1999 Jul, 51(7), 771 - 6
Preparation, relative toxicity and therapeutic efficacy in mice and rats of liposomal HA-1-92, a new oxohexaene polyene macrolide antibiotic; Harindran J et al.; HA-1-92, a new polyene oxohexaene macrolide antibiotic isolated from Streptomyces CDRIL-312, was incorporated into liposomes containing phosphotidyl choline and cholesterol . The liposomal incorporated HA-1-92 considerably decreased toxicity when compared with free HA-1-92 in mice . Liposomal HA-1-92 showed improved pharmacokinetic profiles in rats . When administered to aspergillosis- and cryptococcosis-infected Balb/c mice, liposomal HA-1-92 showed increased antifungal activity, compared with free HA-1-92, with improved survival rate and decreased colony-forming units in lung, liver, spleen and kidney . These results suggest that liposomal HA-1-92 is more effective than free HA-1-92 in controlling experimental aspergillosis and cryptococcosis in Ba1b/c mice.

J Hosp Infect, 1999 Aug, 42(4), 313 - 20
The prognostic factors of cryptococcal meningitis in HIV-negative patients; Lu CH et al.; Seventy-one patients with cryptococcal meningitis, 46 males and 25 females, aged 15-83 years, were included in this study . Their initial clinical manifestations, cerebrospinal fluid (CSF) features, and therapeutic results were analysed . Patients were treated with three different regimens: amphotericin B, fluconazole, and combination therapy . Based on the therapeutic results, the 71 patients were also divided into cured, improved, and failed groups . For statistical comparison, the clinical manifestations and CSF features, were compared according to therapeutic outcome . There was no statistical difference in outcome among the three different antifungal regimens . However, patients treated with fluconazole required 36% fewer days of hospitalization compared with those receiving amphotericin B . Significant prognostic factors, included low CSF glucose, high CSF lactate, high CSF cryptococcal antigen titre (> or = 1:1024), initial level of consciousness, the presence of seizure, hydrocephalus, and central nervous system vasculitis . Multiple logistic regression analysis showed that only initial level of consciousness, and CSF antigen titre were strongly associated with therapeutic failure after other potentially confounding factors were adjusted for . Because some of the prognostic factors in cryptococcal meningitis can be corrected, early diagnosis, early use of appropriate antifungal treatment, and the correction of the underlying metabolic derangements are important in management.

Magn Reson Med, 1999 Sep, 42(3), 442 - 53
Heteronuclear NMR studies of metabolites produced by Cryptococcus neoformans in culture media: identification of possible virulence factors; Bubb WA et al.; The yeast, Cryptococcus neoformans var . neoformans is a major contributor to the morbidity and mortality experienced by the immunosuppressed population . With a view to providing better treatment, identification of cryptococcal virulence factors is an important goal, with most effort to date directed toward the significance of structural variations in the polysaccharide capsule . The present work describes the characterization of supernatants obtained from cryptococcal cultures . This was achieved by thorough identification of the spin systems of individual metabolites through both homonuclear and heteronuclear NMR experiments that circumvented the difficulties imposed by limited dispersion and a range of concentrations in different cultures covering more than 3 orders of magnitude . More than 30 metabolites, including amino acids, alditols, nucleosides, acetate, and ethanol, were identified by their (1)H and (13)C chemical shifts and observed long-range correlations . The possible contribution of some detected substances to the pathogenicity of Cryptococcus neoformans is discussed . Magn Reson Med 42:442-453, 1999 .

J Bacteriol, 1999 Sep, 181(17), 5482 - 8
A unique alpha-1,3 mannosyltransferase of the pathogenic fungus Cryptococcus neoformans; Doering TL; The major virulence factor of the pathogenic fungus Cryptococcus neoformans is an extensive polysaccharide capsule which surrounds the cell . Almost 90% of the capsule is composed of a partially acetylated linear alpha-1,3-linked mannan substituted with D-xylose and D-glucuronic acid . A novel mannosyltransferase with specificity appropriate for a role in the synthesis of this glucuronoxylomannan is active in cryptococcal membranes . This membrane-associated activity transfers mannose in vitro from GDP-mannose to an alpha-1, 3-dimannoside acceptor, forming a second alpha-1,3 linkage . Product formation by the transferase is dependent on protein, time, temperature, divalent cations, and each substrate . It is not affected by amphomycin or tunicamycin but is inhibited by GDP and mannose-1-phosphate . The described activity is not detectable in the model yeast Saccharomyces cerevisiae, consistent with the absence of a similar polysaccharide structure in that organism . A second mannosyltransferase from C . neoformans membranes adds mannose in alpha-1,2 linkage to the same dimannoside acceptor . The two activities differ in pH optimum and cation preference . While the alpha-1,2 transferase does not have specificity appropriate for a role in glucuronoxylomannan synthesis, it may participate in production of mannoprotein components of the capsule . This study suggests two new targets for antifungal drug discovery.

Microbiology, 1999 Aug, 145 ( Pt 8), 1989 - 2000
Secretion of FK506/FK520 and rapamycin by Streptomyces inhibits the growth of competing Saccharomyces cerevisiae and Cryptococcus neoformans; Arndt C et al.; FK506 and rapamycin are immunosuppressants that inhibit signalling cascades required for T-cell activation, yet both are natural products of Streptomyces that live in the soil . FK506 and rapamycin also have potent antimicrobial activity against yeast and pathogenic fungi, suggesting a natural role in inhibiting growth of competing micro-organisms . The immunosuppressive and antimicrobial activities of FK506 and rapamycin are mediated by binding to the FKBP12 prolyl isomerase and the resulting FKBP12/FK506 and FKBP12/rapamycin complexes inhibit conserved protein targets, either the phosphatase calcineurin or the TOR (target of rapamycin) kinases, respectively . Streptomyces sp., 'Streptomyces hygroscopicus subsp . ascomyceticus' and Streptomyces hygroscopicus, which produce FK506, FK520 (also known as ascomycin, a C21 ethyl derivative of FK506) and rapamycin, respectively, produced toxins that inhibited the growth of competing cells of the yeast Saccharomyces cerevisiae and the pathogenic fungus Cryptococcus neoformans . Yeast and fungal mutants lacking FKBP12 or expressing dominant drug-resistant calcineurin or TOR mutants were resistant to FK506 and rapamycin, and to the toxins produced by Streptomyces . Streptomyces strains with mutations in the FK506 or rapamycin biosynthetic enzymes were impaired in toxin production . Finally, the toxins secreted by 'S . hygroscopicus subsp . ascomyceticus' and S . hygroscopicus promoted formation of FKBP12/calcineurin and FKBP12/TOR complexes in a two-hybrid assay and mutations that rendered calcineurin or TOR drug-resistant prevented interaction . These observations support the hypothesis that Streptomyces evolved to secrete FK506, FK520 and rapamycin as toxins to inhibit the growth of competing yeast and fungi.

J Antimicrob Chemother, 1999 Jul, 44(1), 65 - 70
In-vitro interactions of itraconazole with flucytosine against clinical isolates of Cryptococcus neoformans; Barchiesi F et al.; Treatment failures can occur in AIDS patients infected with Cryptococcus neoformans, despite aggressive antifungal therapy . Combination regimens with additive or synergic drugs could provide additional options for treating cryptococcosis . We studied the effects of itraconazole combined with flucytosine against 16 strains of C . neoformans var . neoformans . Combination therapy revealed different results for the various strains, including synergy (fractional inhibitory concentration (FIC) index 0.5, 63% of the interactions), addition (FIC >0.5 to 1.0, 31% of the interactions) and indifference (FIC >1.0 to <2.0, 6% of the interactions) . Antagonism (FIC >2.0) was not observed . The efficacy of combination therapy was confirmed by quantitative cfu and killing curve assays . In particular, killing curves conducted in replicating cells showed that the addition of itraconazole prevented the development of flucytosine-resistant mutants of C . neoformans . These data show that the combination of itraconazole and flucytosine is significantly more active than either drug alone against C . neoformans in vitro.

Curr Opin Microbiol, 1999 Aug, 2(4), 358 - 62
Signal transduction cascades regulating mating, filamentation, and virulence in Cryptococcus neoformans; Wang P et al.; Cryptococcus neoformans is a basidiomycetous fungal pathogen that infects the central nervous system . The organism has a defined sexual cycle involving mating between haploid MATalpha and MATa cells . Recent studies have revealed signaling cascades that coordinately regulate differentiation and virulence of C . neoformans . One signaling cascade involves a conserved G-protein alpha subunit and cAMP, and senses nutrients during mating and virulence . The second is a conserved mitogen activated protein (MAP) kinase cascade that senses pheromone during mating, and also regulates haploid fruiting and virulence . Interestingly, some of the MAP kinase components are encoded by the MAT locus itself, which may explain the unique association of the MATalpha locus with physiology and virulence.

J Acquir Immune Defic Syndr, 1999 Aug 15, 21(5), 401 - 7
Changes in AIDS-defining illnesses in a London Clinic, 1987-1998; Mocroft A et al.; OBJECTIVES: To describe the incidence of AIDS-defining illnesses within a single large clinic setting, to describe temporal changes over a 10-year period in the overall incidence and of individual AIDS-defining illnesses and to investigate the impact of HIV treatment regimen on the incidence of AIDS-defining illnesses . SUBJECTS AND METHODS: A person-years analysis was used to determine the incidence of AIDS-defining illnesses according to calendar year and stratification by CD4 lymphocyte count and treatment regimen in 1806 patients from the Royal Free Centre for HIV Medicine with at least one CD4 lymphocyte count and follow-up visit . RESULTS: Prior to 1992, the incidence of all AIDS-defining illnesses was 27.4/100 person-years of follow-up (PYFU; 95% confidence interval {CI}, 22.8-32.0) and during 1997 this incidence had dropped to 6.9/100 PYFU (95% CI, 4.7-9.1; p < .0001, test for trend) . The decline in incidence over time occurred in esophageal candidiasis, cytomegalovirus disease (including retinitis), Kaposi's sarcoma, lymphoma, wasting syndrome, and Pneumocystis carinii pneumonia (p < .05, test for trend), but there was no evidence of a decline in AIDS dementia or in Mycobacterium avium complex . In 1997, among patients with CD4 lymphocyte counts of < or =200 cells/mm3, the incidence rates for any AIDS-defining illness was 51.1/100 PYFU for patients taking no therapy (95% CI, 27.9-85.7), 34.5 among patients on monotherapy (95% CI, 4.2-124.6), 13.2 among patients taking dual combination therapy (95% CI, 3.6-33.8) and 6.1 among patients taking triple therapy or more complex regimens (95% CI, 0.7-22.0; p < .0001, test for trend) . CONCLUSIONS: There was a considerable decline in AIDS-defining illnesses during 1996 and 1997, coinciding with the rapid development of new antiretroviral treatments and combinations of treatment . Further follow-up of large observational cohorts is essential to monitor the incidence of diagnoses less common than we were able to consider, such as tuberculosis, cryptosporidiosis, and cryptococcosis, and also to investigate whether the incidence of disease continues to fall, or whether it starts to rise again, as toxicities, compliance, drug resistance, and long-term side effects begin to appear.

Infect Immun, 1999 Sep, 67(9), 4620 - 7
The capsule of Cryptococcus neoformans reduces T-lymphocyte proliferation by reducing phagocytosis, which can be restored with anticapsular antibody; Syme RM et al.; Cell-mediated immunity is critical for the host defense to Cryptococcus neoformans, as demonstrated by numerous animal studies and the prevalence of the infection in AIDS patients . Previous studies have established that the polysaccharide capsule contributes to the virulence of C . neoformans by suppressing T-lymphocyte proliferation, which reflects the clonal expansion of T lymphocytes that is a hallmark of cell-mediated immunity . The present studies were performed to identify the major mechanism by which polysaccharide impairs lymphocyte proliferation, since capsular polysaccharide has the potential to affect the development of T-lymphocyte responses by stimulating production of interleukin-10 (IL-10), inhibiting phagocytosis, and inducing shedding of cell surface receptors . We demonstrate that polysaccharide inhibits lymphocyte proliferation predominantly by blocking uptake of C . neoformans, which is crucial for subsequent lymphocyte proliferation . In addition, we show that polysaccharide did not suppress lymphocyte proliferation via an IL-10-dependent mechanism, nor did it affect critical surface receptor interactions on the T cell or antigen-presenting cell . Having established that polysaccharide impairs phagocytosis, we performed studies to determine whether opsonization with human serum or with anticapsular antibody could reverse this effect . Impaired uptake and lymphocyte proliferation that were induced by polysaccharide can be enhanced through opsonization with monoclonal antibodies or human serum, suggesting that antipolysaccharide antibodies might enhance the host defense by restoring uptake of the organism and subsequent presentation to T lymphocytes . These studies support the therapeutic potential of stimulating cell-mediated immunity to C . neoformans with anticapsular antibody.

Infect Immun, 1999 Sep, 67(9), 4469 - 76
Molecular and idiotypic analyses of the antibody response to Cryptococcus neoformans glucuronoxylomannan-protein conjugate vaccine in autoimmune and nonautoimmune mice; Nussbaum G et al.; The antibody response to Cryptococcus neoformans capsular glucuronoxylomannan (GXM) in BALB/c mice frequently expresses the 2H1 idiotype (Id) and is restricted in variable gene usage . This study examined the immunogenicity of GXM-protein conjugates, V (variable)-region usage, and 2H1 Id expression in seven mouse strains: BALB/c, C57BL/6, A/J, C3H, NZB, NZW, and (NZB x NZW)F(1) (NZB/W) . All mouse strains responded to vaccination with GXM conjugated to tetanus toxoid (TT), the relative magnitude of the antibody response being BALB/c approximately C3H > C57BL/6 approximately NZB approximately NZW approximately NZB/W > A/J . Analysis of serum antibody responses to GXM with polyclonal and monoclonal antibodies to the 2H1 Id revealed significant inter- and intrastrain differences in idiotype expression . Thirteen monoclonal antibodies (MAbs) (two immunoglobulin M {IgM}, three IgG3, one IgG1, three IgG2a, two IgG2b, and two IgA) to GXM were generated from one NZB/W mouse and one C3H/He mouse . The MAbs from the NZB/W mouse were all 2H1 Id positive (Id(+)) and structurally similar to those previously generated in BALB/c mice, including the usage of a V(H) from the 7183 family and Vkappa5.1 . Administration of both 2H1 Id(+) and Id(-) MAbs from NZB/W and C3H/H3 mice prolonged survival in a mouse model of cryptococcosis . Our results demonstrate (i) that V-region restriction as indicated by the 2H1 Id is a feature of both primary and secondary responses of several mouse strains; and (ii) that there is conservation of V-region usage and length of the third complementarity-determining region in antibodies from three mouse strains . The results suggest that V-region restriction is a result of antibody structural requirements necessary for binding an immunodominant antigen in GXM.

Yeast, 1999 Aug, 15(11), 1133 - 9
Molecular cloning of homologs of RAS and RHO1 genes from Cryptococcus neoformans; Tanaka K et al.; We cloned and sequenced homologs of RAS(CnRAS) and RHO1(CnRHO1) genes from Cryptococcus neoformans . The proteins encoded by the CnRAS and CnRHO1 genes contained 216 and 197 amino acids, respectively . The deduced amino acid sequence of the CnRAS gene shared a high degree of sequence identity with the Ras proteins in other fungal species: Coprinus cinereus(76%), Lentinula edodes(74%), Saccharomyces cerevisiae RAS2(72%), and Schizosaccharomyces pombe(68%) . The deduced amino acid sequence of the CnRHO1 gene shared a high degree of sequence identity with the Rho1 proteins in other fungal species: Candida albicans(78%), S . pombe(77%) and S . cerevisiae(76%) . The deduced proteins contained GTP-binding and GTP-hydrolysis domains, and the prenylation site that are conserved among the small G protein superfamily . The synthetic peptides that contained the C-terminal amino acid sequence of the CnRas and CnRho1 proteins were geranylgeranylated .

Planta Med, 1999 Jun, 65(5), 444 - 6
Evaluation of the antimicrobial potency of tannins and related compounds using the microdilution broth method; Kolodzeij H et al.; The antimicrobial activity of a total of 27 tannins and related compounds was evaluated against 8 microorganisms, including 2 Gram-positive (Bacillus subtilis, Staphylococcus aureus), 4 Gram-negative bacteria (Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Proteus mirabilis), and 2 yeasts (Candida albicans, Cryptococcus neoformans) . The compounds tested were generally found to possess only weak to moderate antibacterial, but fairly high anticryptococcal activities . Attention is given to structure-activity relationships with emphasis on simple galloyl esters, hydrolyzable tannins and proanthcyanidins among this class of secondary products.

Clin Infect Dis, 1999 May, 28(5), 1049 - 56
Itraconazole prophylaxis for fungal infections in patients with advanced human immunodeficiency virus infection: randomized, placebo-controlled, double-blind study . National Institute of Allergy and Infectious Diseases Mycoses Study Group; McKinsey DS et al.; In a prospective, randomized, double-blind trial, 149 patients with advanced human immunodeficiency virus (HIV) infection were randomized to receive itraconazole capsules (200 mg daily) and 146 to receive a matched placebo . Both groups were monitored for evidence of fungal infections . Baseline characteristics of the two groups were similar . Failure of prophylaxis occurred in 29 (19%) of the itraconazole recipients and 42 (29%) of the placebo recipients (P = .004; log-rank test) . There were 6 invasive fungal infections in the itraconazole group (4, histoplasmosis; 1, cryptococcosis; 1, aspergillosis) and 19 in the placebo group (10, histoplasmosis; 8, cryptococcosis; 1, aspergillosis) (P = .0007; log-rank test) . Itraconazole significantly delayed time to onset of histoplasmosis (P = .03; log-rank test) and cryptococcosis (P = .0005; log-rank test) . Prophylaxis failure due to recurrent or refractory mucosal candidiasis occurred with similar frequency in the two groups (itraconazole, 15%; placebo, 16%) . A survival benefit was not demonstrated . Itraconazole generally was well tolerated . Primary prophylaxis with itraconazole capsules prevents histoplasmosis and cryptococcosis in patients with HIV infection.

J Med Microbiol, 1999 Aug, 48(8), 731 - 40
Biochemical and functional characterisation of secreted phospholipase activities from Cryptococcus neoformans in their naturally occurring state; Santangelo RT et al.; A recent study demonstrated that phospholipase B (PLB), lysophospholipase (LPL) and lysophopholipase transacylase (LPTA) are secreted by Cryptococcus neoformans var . neoformans and showed that the amount of enzyme production correlated with virulence in mice . The present study characterised the extracellular enzyme activities further by radiometric assays and 31P nuclear magnetic resonance spectroscopy (NMR) . All three enzymes were most active between 25 and 40 degrees C . Bovine lung surfactant and its major lipid components, disaturated phosphatidylcholine and phosphatidylglycerol, were the optimal substrates for PLB . Lysophosphatidylcholine was the favoured substrate for LPL and LPTA . PLB and LPL/LPTA were differentially affected by Triton X-100, and palmitoyl carnitine was a potent inhibitor of the three phospholipases . LPL and PLB activities were inhibited by dithiothreitol; N-ethylmaleimide inhibited LPL and LPTA activities . None of the enzymes was inhibited by N-bromosuccinimide or p-bromophenacyl bromide . Cellular disruption experiments indicated that >85% of the phospholipase activities were cell-associated, with LPL and LPTA being more easily released than PLB . At pH 5.5 and 7.0, the heat-inactivated secreted enzyme preparations decreased the viability of human neutrophils . This effect was attenuated by active supernates . The relative activities of the PLB, LPL and LPTA in the environment of neutrophils are likely to determine the fate of these cells in vivo . Both phospholipases and heat-stable substances secreted by C . neoformans at 37 degrees C could contribute to membrane degradation and virulence.

J Clin Microbiol, 1999 Sep, 37(9), 2920 - 6
Presence of alpha and a mating types in environmental and clinical collections of Cryptococcus neoformans var . gattii strains from Australia; Halliday CL et al.; Cryptococcus neoformans var . gattii lives in association with certain species of eucalyptus trees and is a causative agent of cryptococcosis . It exists as two mating types, MATalpha and MATa, which is determined by a single-locus, two-allele system . In the closely related C . neoformans var . neoformans, the alpha mating type has been found to outnumber its a counterpart by at least 30:1, but there have been very limited data on the proportions of each mating type in C . neoformans var . gattii . In the present study, specific PCR primers were designed to amplify two separate alpha-mating-type genes from C . neoformans var . gattii strains . These were used to survey for the presence of the two mating types in clinical and environmental collections of C . neoformans var . gattii strains from Australia . Sixty-eight of 69 clinical isolates produced both alpha mating type-specific bands and were assumed to be of the alpha mating type . The majority of environmental isolates were also of the alpha mating type, but the a mating type was located in two separate areas . In one area, the a mating type outnumbered the alpha mating type by 27:2, but in the second area, the ratio of the two mating types was close to the 50:50 ratio expected for sexual recombination.

Farmaco, 1999 Jun 30, 54(6), 416 - 22
Synthesis and biological evaluation of {alpha-(1,5-disubstituted 1H-pyrazol-4-yl)benzyl}azoles, analogues of bifonazole; Menozzi G et al.; A series of pyrazole analogues of bifonazole, an antifungal drug used in clinical practice, 2a-h and 4a-h were synthesized and tested in vitro against Candida albicans, Cryptococcus neoformans and Aspergillus fumigatus, with no significant results . Imidazoles 2a-h were also tested in vivo for antiarrhythmic and antihypertensive activities; two of these compounds showed moderate activity against ventricular fibrillation caused by aconitine in rats . The above compounds were prepared by reaction of phenyl-{5 substituted 1-phenyl (or 1-methyl)-1H-pyrazol-4-yl}methanols with N,N'-carbonyldiimidazole (2a-h) or of the respective chloro derivatives with 1H-1,2,4-triazole (4a-h).

Eur J Clin Microbiol Infect Dis, 1999 Jun, 18(6), 399 - 402
Rare AIDS-defining diseases in the Swiss HIV Cohort Study; Burckhardt B et al.; The objective of this study was to investigate the spectrum and frequency of rare AIDS-defining diseases in the Swiss HIV Cohort Study . AIDS-defining diseases contributing less than 1% to the absolute number of all recorded AIDS-defining diseases in at least one of five periods (1988-1990, 1991-1992, 1993-1994, 1995-1996, 1997) were defined as being rare . A total of 9110 HIV-infected subjects were included in this study . Over the entire 9-year period, the following rare diseases were diagnosed: progressive multifocal leukoencephalopathy (n = 138), disseminated cryptococcosis (n = 67), visceral herpes simplex disease (n = 66), primary cerebral lymphoma (n = 65), indeterminate cerebral lesion (n = 50), cryptococcal meningitis (n = 34), Mycobacterium kansasii disease (n = 32), recurrent Salmonella septicemia (n = 22), intestinal isosporiasis (n = 21), candidiasis of the trachea, bronchi and lungs (n = 19), toxoplasma retinitis (n = 16), disseminated toxoplasmosis (n = 8), invasive cervical carcinoma (n = 8), extrapulmonary Pneumocystis disease (n = 5), disseminated histoplasmosis (n = 1) and disseminated coccidioidomycosis (n = 1) . Rare diseases accounted for 7.3% of all AIDS-defining diseases over the entire 9-year period . Physicians should be aware of the likelihood of a broad spectrum of AIDS-defining diseases in HIV-infected patients.

Enferm Infecc Microbiol Clin, 1999 Jun-Jul, 17(6), 274 - 8
{Cryptococcosis not associated with AIDS . Clinical study of seven patients}; Pintado V et al.; BACKGROUND: The aim of the present was to study the clinical characteristics, the diagnostic methods, response to treatment and the evolution of cryptococcosis not associated with AIDS . METHODS: Seven cases studied in the Hospital Ramon y Cajal of Madrid, Spain over a 19-year period (1980-1998) . Diagnosis was determined by LCR culture, blood or other clinical samples . RESULTS: Four patients were males with a mean age of 45.1 years (range: 23-69 years) . All except one case presented a predisposing disease and three received corticoids and/or immunosuppressive drugs . Six patients presented meningitis and one pneumonia . The meningitis showed a subacute course with pleocytosis, hypoglycorrhachia and hyperproteinorrhachia in most of the patients . Chinese ink staining, culture and the cryptococcic antigen of LCR were positive in all the cases of meningitis evaluated . All the patients were initially treated with amphotericin and fluorocytosin . This schedule was substituted in two cases by amphotericin lipid formulas because of adverse effects and by fluconazol as an elective choice in one case . The evaluation was favorable in all the patients . One patient presented meningeal and cutaneous recurrence . Three patients died, although none did so during the acute episode or as a consequence of the infection . CONCLUSIONS: Cryptococcosis not associated with AIDS is rare in Sapin . It usually evolves with subacute hypoglucorrhachic meningitis and the conventional diagnostic techniques (Chinese ink staining, cryptococcic antigen and culture) provide adequate profitability . Response to treatment with amphotericin and fluorocytosis is usually favorable, although there are therapeutic alternatives which are adequate for selected cases.

J Infect Dis, 1999 Sep, 180(3), 915 - 9
Antibodies reactive with the cryptococcal capsular polysaccharide glucuronoxylomannan are present in sera from children with and without human immunodeficiency virus infection; Abadi J et al.; Antibodies reactive with the cryptococcal polysaccharide glucuronoxylomannan (GXM) are present in sera from both human immunodeficiency virus (HIV)-uninfected and -infected adults . However, the prevalence of these antibodies in US children is unknown . An antigen-capture ELISA was used to determine the presence, serotype specificity, isotype, and IgG subclass distribution of antibodies to GXM in sera from 27 HIV-uninfected and 34 HIV-infected children . The children were of similar age and socioeconomic background . IgG and IgM to GXM were present in sera from all children, although HIV-uninfected children had significantly higher titers . HIV-uninfected children had IgG1 and IgG2 to GXM and significantly greater IgG2 than in HIV-infected children . These findings of type-specific antibodies to GXM in early childhood suggest that exposure to or subclinical infection with Cryptococcus neoformans may be an early childhood event.

J Infect Dis, 1999 Sep, 180(3), 791 - 801
Monoclonal antibody-mediated toxicity in Cryptococcus neoformans infection: mechanism and relationship to antibody isotype; Lendvai N et al.; Antibody reagents represent an alternative for the therapy of human cryptococcosis, and monoclonal antibody 18B7 (IgG1) is a candidate for phase I trial in humans with cryptococcosis . However, antibody administration to mice with established Cryptococcus neoformans infection has been reported to produce acute lethal toxicity (ALT) . The present study confirmed this phenomenon and investigated the mechanism of ALT . ALT was associated with hemoconcentration, hypotension, and circulatory collapse; however, toxicity could be prevented by platelet-activating factor inhibitor, rat antibody to Fc receptor, or IgM before IgG1 . Significant isotype-specific differences were found in ALT, which can be interpreted as consistent with the hypothesis that there are distinct Fc receptors for murine IgG1 and IgG3 . The IgM and IgG3 isotype preference in antibody responses to polysaccharide antigens in mice may translate to a lack of toxicity of antigen-antibody complexes during the course of infections with encapsulated pathogens.

J Am Optom Assoc, 1999 Jun, 70(6), 384 - 90
Sarcoidosis-related anterior uveitis in a patient with human immunodeficiency virus; Lee AK et al.; BACKGROUND: This is the first ophthalmic report--to our knowledge--of an anterior uveitis secondary to sarcoidosis in a patient infected with human immunodeficiency virus (HIV) . Other reported causes of uveitis in HIV-infected patients have included HIV, herpes zoster, tuberculosis, syphilis, toxoplasmosis, cryptococcus, rifabutin prophylaxis for mycobacterium, and protease inhibitors such as ritonavir and indinavir . Uveitis secondary to sarcoidosis in the non-HIV population is classically seen in young, female, African-American patients . There are rare reports, found exclusively in the pulmonary literature, of sarcoidosis in HIV-infected patients . CASE REPORT: A 38-year-old African-American male infected with HIV was treated for chronic recurrent anterior uveitis secondary to sarcoidosis . His sarcoidosis was diagnosed 1 month earlier, along with the onset of his uveitis . During the previous 6 years he has been treated with anti-HIV antivirals as well as prophylaxis for opportunistic infections . To date, his infectious disease specialist continues to treat his HIV and systemic sarcoidosis . CONCLUSION: Patients with HIV infection in whom sarcoidosis with secondary uveitis develops are very rare . Management of these patients requires careful use of topical and oral steroidal anti-inflammatories to control ocular and systemic sequelae of sarcoidosis . This case initiates some interesting questions about the immunology of sarcoidosis and its presence in immunocompromised patients . Use of steroids in an immunocompromised patient is clinically complex . Further clinical study is needed to elicit the full clinical significance of sarcoidosis and HIV infection.

Rev Inst Med Trop Sao Paulo, 1999 Jan-Feb, 41(1), 27 - 32
Fever of undetermined origin in patients with the acquired immunodeficiency syndrome in Brazil: report on 55 cases; Lambertucci JR et al.; The medical records of patients with AIDS admitted to a general hospital in Brazil from 1989 to 1997 were reviewed retrospectively with the aim at defining the frequency and etiology of fever of undetermined origin (FUO) in HIV-infected patients of a tropical country and to evaluate the usefulness of the main diagnostic procedures . 188 (58.4%) out of 322 patients reported fever at admission to hospital and 55 (17.1%) had FUO . Those with FUO had a mean CD4+ cell count of 98/ml . A cause of fever was identified for 45 patients (81.8%) . Tuberculosis (32.7%), Pneumocystis carinii pneumonia (10.9%), and Mycobacterium avium complex (9.1%) were the most frequent diagnoses . Other infectious diseases are also of note, such as cryptococcal meningitis (5.5%), sinusitis (3.6%), Salmonella-S . mansoni association (3.6%), disseminated histoplasmosis (3.6%), neurosyphilis (1.8%), and isosporiasis (1.8%) . Four patients had non-Hodgkin's lymphoma (7.3%) . We conclude that an initial aggressive diagnostic approach should be always considered because biopsies (lymph node, liver and bone marrow) produced the highest yield in the diagnosis of FUO and the majority of the diagnosed diseases are treatable . The association of diseases is common and have contributed to delay the final diagnosis of FUO in most cases . In our study area the routine request of hemocultures for Salmonella infection and the investigation of cryptococcal antigen in the serum should be considered.

Electrophoresis, 1999 Jun, 20(8), 1790 - 9
Molecular typing of global isolates of Cryptococcus neoformans var . neoformans by polymerase chain reaction fingerprinting and randomly amplified polymorphic DNA-a pilot study to standardize techniques on which to base a detailed epidemiological survey; Meyer W et al.; A total of 356 clinical isolates of the encapsulated basidiomycetous fungus Cryptococcus neoformans var . neoformans, obtained from Australia, Argentina, Brazil, India, Italy, New Zealand, Papua New Guinea, South Africa, Thailand and the USA, were analyzed to lay the basis for a comprehensive evaluation of the global genetic structure of C . neoformans . Two polymerase chain reaction (PCR)-based typing techniques were standardized: PCR fingerprinting using a single primer specific to minisatellite or microsatellite DNA, and randomly amplified polymorphic DNA (RAPD) analysis using two combinations of three 20- to 22-mer random primers . Previous studies showed that the resultant profiles are reproducible and stable over time . Identical results were obtained in two different laboratories and by different scientists in the same laboratory . Both typing techniques separated the isolates into four major groups (VNI and VNII, serotype A; VNIII, serotype A/D; and VNIV, serotype D) . The majority (78%) of isolates belonged to VNI, compared with 18% VNII, 1% VNIII and 3% VNIV . All US isolates could be differentiated by a unique, strain-specific PCR fingerprint or RAPD pattern in contrast to most of the non-US isolates, which showed a substantially higher degree of genetic homogeneity, with some clonality, in different parts of the world . Isolates obtained from the same patient at different times and from different body sites, had identical banding patterns . Both typing techniques should provide powerful tools for epidemiological studies of medically important fungi.

J Chemother, 1999 Jun, 11(3), 187 - 90
Comparative study of the in vitro antifungal activity of bifonazole, naftifine and sertaconazole against yeasts; Carrillo-Munoz AJ et al.; The in vitro activity of three antifungal agents was tested and compared against 151 yeast strains, including ten Candida species, Cryptococcus neoformans, Rhodotorula rubra, and Trichosporon cutaneum . Minimum inhibitory concentrations (MICs) were determined by a microdilution technique in Shadomy modified liquid medium . The mean MICs of sertaconazole (0.34 mg/L) were lower than those of naftifine (16.3 mg/L) and bifonazole (13.2 mg/L) . These results suggest that sertaconazole is more active against Candida spp than other topical agents such as bifonazole and naftifine.

Intern Med, 1999 Jul, 38(7), 563 - 9
In situ detection of Aspergillus 18S ribosomal RNA in invasive pulmonary aspergillosis; Kobayashi M et al.; OBJECTS: We attempted to evaluate the usefulness of in situ hybridization (ISH) in the specific diagnosis of Aspergillus pulmonary infection . METHODS: We used an ISH technique using a multiple digoxigenin-incorporating probe, which was constructed by means of the polymerase chain reaction (PCR) from the 18S ribosomal RNA of Aspergillus fumigatus . MATERIALS: We studied twelve formalin-fixed, paraffin-embedded lung tissue sections from autopsy-confirmed invasive pulmonary aspergillosis (IPA) (5 acute myelocytic leukemias, 2 acute lymphocytic leukemias, 2 chronic myelocytic leukemias, 1 adult T-cell leukemia, 1 non-Hodgkin's lymphoma and 1 chronic obstructive pulmonary disease.), and 18 sections from other pulmonary infections as control . RESULTS: ISH using the probe and a low-viscosity hybridization buffer solution (LV) positively stained hyphal elements in 12 of 12 autopsy lung tissue specimens from subjects with IPA, while ISH using the probe and a high viscosity hybridization buffer solution (HV) positively stained the hyphal elements in 6 of 12 . Specifically, ISH (LV) demonstrates hyphal elements of Aspergillus spp . in the center of Aspergillus abscess . While, ISH (HV) can detect hyphal elements located in the periphery of a suppurative abscess as well as those in the blood vessel . Conversely, ISH did not show positive results for any of the autopsy tissue specimens from subjects with other fungal pneumonia infections (Candida n=5, Mucor n=2, Cryptococcus n=2, and Pseudallescheria n=1), Pneumocystis carinii pneumonia (n=5), and cytomegalovirus pneumonia (n=3) . Dual staining by means of ISH and immunohistochemistry (IHC) using anti-neutrophil elastase (NE) and anti-CD68 monoclonal antibodies showed that NE positive cells were localized at the edge of the radial growth of the organism, but CD68 positive cells were located around the center of the abscess . The accumulation of NE positive cells was rarely seen in half of the cases (6/12) . In contrast, CD68 positive cells were routinely present in the center of the abscess (12/12) . CONCLUSION: ISH in conjunction with IHC is a useful tool for differentiating Aspergillus spp . from other fungal genera in tissue sections from patients with IPA and may have a certain role in the evaluation of the interactions between organisms and recruiting inflammatory cells.

Rinsho Byori, 1999 Jun, 47(6), 487 - 93
{Non-culture based laboratory diagnosis of sepsis}; Obayashi T; Blood culture is clearly the most important diagnostic procedure for sepsis . In the majority of cases, however, it yields negative results when bacterial or fungal sepsis is strongly suspected in view of an excellent response to antibacterial or antifungal medication . This is likely because infecting microbes have already been too seriously damaged in the blood stream to grow in culture media . Therefore, various tests have been devised to detect cellular components of pathogens including endotoxin for gram-negative bacteria, (1-->3)-beta-D-glucan for fungi, a non-mannan heat-labile candida antigen, glucronoxylomannan for Cryptococcus neoformans, and galactomannan for Aspergillus . The first two tests are particularly suitable for screening purposes, because, in addition to their high sensitivity and specificity, they cover a wide range of species in each category, though not as widely as blood culture . The candida antigen detectable by Candi-Tec is probably a complex of immune-related proteins, and appears to be a poor indicator in immunocompromised hosts . Glucronoxylomannan has been established as a useful marker of cryptococcosis, whereas galactomannan, though highly specific for aspergillosis, needs a more sensitive detection system to be useful for diagnostic purposes.

Hepatogastroenterology, 1999 May-Jun, 46(27), 1618 - 22
Cryptococcal peritonitis: report of a case and review of the literature; Stiefel P et al.; We describe a patient diagnosed with AIDS and cirrhosis who had recently suffered a self-limited and non-specific esophageal ulceration . After this, he was hospitalized because of an oral bleeding with fatal evolution, and Cryptococcus neoformans was isolated from ascitic fluid during a routine paracenteses . We have reviewed the literature and, since 1963, only another 10 cases of cryptococcal peritonitis have been reported . A liver disease and not the AIDS (surprisingly, our case is the only report of cryptococcal peritonitis in a subject having both diseases) was the most common underlying disease (72.7%) and was associated with the worst prognosis (only one patient survived) . An oral or upper gastrointestinal bleeding was the most common associated circumstance although recent steroid or antibiotic therapy has been also reported . Finally, diagnosis was delayed in many patients . The reasons for these delays are discussed.

Antimicrob Agents Chemother, 1999 Aug, 43(8), 1975 - 81
A novel injectable water-soluble amphotericin B-arabinogalactan conjugate; Falk R et al.; New, stable, highly water-soluble, nontoxic polysaccharide conjugates of amphotericin B (AmB) are described . AmB was conjugated by a Schiff-base reaction with oxidized arabinogalactan (AG) . AG is a highly branched natural polysaccharide with unusual water solubility (70% in water) . A high yield of active AmB was obtained with the conjugates which were similarly highly water soluble and which could be appropriately formulated for injection . They showed comparable MICs for Candida albicans and Cryptococcus neoformans (MICs, 0.1 to 0.2 microg/ml) . The reduced AmB conjugate, which was synthesized at pH 11 for 48 h at 37 degrees C, was nonhemolytic and was much safer than conventional micellar AmB-deoxycholate . It was the least toxic AmB-AG conjugate among those tested with mice (maximal tolerated dose, 50 mg/kg of body weight), and histopathology indicated no damage to the liver or kidneys . This conjugate, similarly to the liposomal formulation (AmBisome), was more effective than AmB-deoxycholate in prolonging survival . It was more effective than both the liposomal and the deoxycholate formulations in eradicating yeast cells from target organs . The overall results suggest that after further development of the AmB-AG conjugate, it may be a potent agent in the treatment of fungal infections.

Antimicrob Agents Chemother, 1999 Aug, 43(8), 1856 - 61
Heteroresistance to fluconazole and voriconazole in Cryptococcus neoformans; Mondon P et al.; Cryptococcus neoformans isolates that exhibited unusual patterns of resistance to fluconazole and voriconazole were isolated from seven isolates from two different geographical regions: one isolate from an Israeli non-AIDS patient and six serial isolates from an Italian AIDS patient who had suffered six recurrent episodes of cryptococcal meningitis . Each isolate produced cultures with heterogeneous compositions in which most of the cells were susceptible, but cells highly resistant to fluconazole (MICs, >/=64 microg/ml) were recovered at a variable frequency (7 x 10(-3) to 4.6 x 10(-2)) . Evidence showed that this type of resistance is innate and is unrelated to drug exposure since the Israeli patient had never been treated with azoles or any other antimycotic agents . Analysis of clonal subpopulations of these two strains showed that they exhibited heterogeneous patterns of resistance . The number of subpopulations which grew on fluconazole or voriconazole agar declined progressively with increasing azole concentration without a sharp cutoff point . For the Italian serial isolates, the number of clonal populations resistant to fluconazole (64 microg/ml) and voriconazole (1 microg/ml) increased steadily, yielding the highest number for the isolate from the last episode . Attempts to purify a sensitive subpopulation failed, but clones highly resistant to fluconazole (100 microg/ml) and moderately resistant to voriconazole (1 microg/ml) always produced a homogeneous population of resistant cells . Upon maintenance on drug-free medium, however, the majority of the homogeneously resistant cells of these subclones lost their resistance and returned to the stable initial heteroresistant phenotype . The pattern of heteroresistance was not affected by the pH or osmolarity of the medium but was influenced by temperature . The resistance appeared to be suppressed at 35 degrees C and was completely abolished at 40 degrees C . Although heterogeneity in azole resistance among subpopulations of single isolates has been reported for Candida species, the transient changes in expression of resistance under different growth conditions reported here have not been observed in fungal pathogens.

J Acquir Immune Defic Syndr, 1999 Aug 1, 21(4), 326 - 32
Clinical presentation of hospitalized adult patients with HIV infection and AIDS in Bangkok, Thailand; Tansuphasawadikul S et al.; OBJECTIVE: To characterize the clinical spectrum of disease and immune status of adult HIV-1-infected patients in Bangkok . DESIGN: Cross-sectional survey of hospital admissions . METHODS: From November 1993 through June 1996, demographic, clinical, and laboratory data were collected from HIV-infected inpatients (> or =14 years old) at an infectious diseases hospital . RESULTS: Of 16,717 persons admitted, 3112 (18.6%) were HIV-seropositive, 2261 of whom were admitted for the first time . Of 2261, 1926 (85.2%) were male, 1942 (85.9%) had been infected heterosexually or by means not related to drug use, 319 (14.1%) were injection drug users (IDUs), and 1553 (68.7%) had AIDS . The most common AIDS-defining conditions were extrapulmonary cryptococcosis (EPC; 38.4%), tuberculosis (TB; 37.4%), and wasting syndrome (WS; 8.1%) . IDUs were more likely (p < .05) to have TB or WS but less likely (p < .05) to have EPC or Pneumocystis carinii pneumonia than patients with no history of injection drug use . Lymphocyte counts were measured for 2047 (90.5%) patients; 81.8% had < or =1500 lymphocytes/microl . CONCLUSION: These HIV-infected patients were admitted with severe immunosuppression . Cryptococcosis and TB are major problems and differ in prevalence among IDUs and persons infected sexually . Clinical and immunologic information is critical in improving the lives of HIV-infected persons in Asia through prevention, treatment, and prophylaxis.

J Nat Prod, 1999 Jul, 62(7), 988 - 92
Microbial transformation of sampangine; Orabi KY et al.; Microbial transformation studies of the antifungal alkaloid sampangine (2) have revealed that it is metabolized by a number of microorganisms . Using a standard two-stage fermentation technique, Beauvaria bassiana (ATCC 7159), Doratomyces microsporus (ATCC 16225), and Filobasidiella neoformans (ATCC 10226) produced the 4'-O-methyl-beta-glucopyranose conjugate (3), while Absidia glauca (ATCC 22752), Cunninghamella elegans (ATCC 9245), Cunninghamella species (NRRL 5695), and Rhizopus arrhizus (ATCC 11145) produced the beta-glucopyranose conjugate (4) . Metabolites 3 and 4 have been characterized on the basis of spectral data . Both 3 and 4 had significant in vitro activity against Cryptococcus neoformans but were inactive against Candida albicans . Metabolite 4 was inactive in vivo in a mouse model of cryptococcosis.

Mycoses, 1999, 42(4), 231 - 7
Comparative evaluation of Fungitest-, Neo-Sensitabs- and M27T-NCCLS broth microdilution methods for antifungal drug susceptibility testing of Candida species and Cryptococcus neoformans; Swinne D et al.; Two commercial antifungal susceptibility testing systems (Fungitest and Neo-Sensitabs) were compared with the M27T-NCCLS reference broth microdilution method using one hundred isolates of Candida sp . and Crptococcus neoformans . Six different antifungal drugs were tested: amphotericin B, 5-fluorocytosine, fluconazole, itraconazole, ketoconazole and miconazole . The overall agreement between the Fungitest and the reference methods was much better than between the Neo-Sensitabs and the reference methods: the agreement for the Fungitest ranged from 100% for amphotericin B to 76.7% for itraconazole whereas for the Neo-Sensitabs, it ranged from 90.4% for amphotericin B to 36% for ketoconazole . For the total number of tests performed with Neo-Sensitabs, there were 37.8% of discrepancies with the reference method whereas for the tests performed with Fungitest, there was only 16.5% of discrepancies . Major discrepancies, defined as results that classified an isolate as susceptible by one method and resistant by another, occurred in 21 cases for the Neo-Sensitabs test and only in four cases with the Fungitest, namely 0.6% of the cases . We conclude that the Fungitest method constitutes a simple and reliable procedure for antifungal drug susceptibility testing.

Nippon Ishinkin Gakkai Zasshi, 1999, 40(3), 143 - 9
{Controversial points in the treatment of patients with haematologic malignancies complicated with systemic fungal infections}; Mori T; Prophylaxis and treatments for fungal infections differ with the infection type . However, the eradication of risk factors for outbreak of fungal infections, and the usage of appropriate antifungal agents are universally important to prevent these infections . For infections due to intrinsic fungi such as Candida spp., risk factors such as changes in normal flora by aggressive and prolonged broad-spectrum antibiotics therapy should not be permitted to emerge . On the other hand, infections due to extrinsic fungi such as Cryptococcus neoformans and Aspergillus spp . can be prevented by eradication of the colonized fungi using antifungal prophylaxis and the use of air-cleaning machines to combat colonization of patients and contamination in the hospital environment . The most important risk factor of fungal infections in patients suffering from haematologic malignancies is leukopenia . In these cases, it is crucial to reduce the duration of neutropenia and enhance the anti-microbial function using granulocyte-macrophage-colony stimulating factor . When a patient is complicated with a fungal infection, appropriate antifungal agents must be used at appropriate dosages for the appropriate period . However, there are still very few satisfactory antifungals with minimal adverse effects and good potential efficacy for systemic fungal infections . Therefore, combination therapy with amphotericin B and azole antifungals is necessary for patients with severe fungal infections . In patients complicated with fungal infections, the underlying disease is often resistant to aggressive antifungal therapy . Control of this underlying disease is thus a most important therapeutic factor.

Nippon Ishinkin Gakkai Zasshi, 1999, 40(3), 157 - 61
{The future of antifungal agents . Non azole antifungal agents}; Maesaki S et al.; We investigated the efficacy of non-azole antifungal agents . Long circulating immunoliposomal amphotericin B was potent in murine invasive pulmonary aspergillosis . The concentration of AMPH-B was still high in the lung after 6 hours of 34A-PEG-liposomal AMPH-B . Lipid nanosphere amphotericin B (NS-718) showed efficacy against pulmonary aspergillosis in rats and pulmonary cryptococcosis in mice . The renal toxicity of NS-718 was estimated to be lower than that of AMPH-B from the results of the toxicity study in the rat infusion model . FK 463, a novel (1,3)-beta-D-glucan synthase inhibitor, showed efficacy against azole-resistant Candida albicans in murine experimental disseminated candidiasis . FK463 could be a promising drug and the therapy of choice for azole resistant C . albicans infection.

Mycopathologia, 1998-99, 144(1), 21 - 7
In vitro effect of lung surfactant on alveolar macrophage defence mechanisms against Cryptococcus neoformans; Gross NT et al.; The effects of a modified natural porcine surfactant (Curosurf) on phagocytosis and killing of Cryptococcus neoformans by alveolar macrophages and on the production of superoxide anions were investigated in vitro . Attachment and ingestion were evaluated separately by a fluorescent quenching technique . The nitroblue tetrazolium reduction test was used as an indirect measurement of superoxide anion production . Killing was assessed by a colony-forming assay . Surfactant induced increased ingestion of C . neoformans, unopsonized as well as opsonized with fresh serum or anticryptococcal polyclonal IgG . Surfactant had, however, no effect on the attachment or killing of unopsonized or opsonized C . neoformans by the alveolar macrophages . In addition, the enhancement of the oxidative metabolism of the macrophages after stimulation with opsonized yeast was impaired, although the killing was not affected . This study indicates that in vitro Curosurf can influence the alveolar macrophage defence against C . neoformans by enhancing its ingestion and by interacting with the superoxide anions release from alveolar macrophages stimulated with fresh serum or anticryptococcal polyclonal IgG opsonized yeast cells.

Mycopathologia, 1998-99, 144(1), 1 - 7
Isolation of a human serum protein that inhibits the growth of Cryptococcus neoformans; Duvvuru S et al.; Human serum at 5 to 10% (v/v) in tissue culture medium RPMI-1640, inhibits the growth of Cryptococcus neoformans by 80 to 93% . Serum fractionated on molecular sieve columns (Sephadex G-200) yielded an active protein fraction . This fraction at 100 micrograms protein/ml inhibited the growth of C . neoformans by 54% . When an active G-200 fraction was applied to a dye affinity column (Affi-Gel Blue) the fraction with inhibitory activity was bound by the column and was eluted with 1.4 M NaCl in 0.1 M phosphate buffer (pH 7.4) . The bound fraction at 62.5 micrograms protein/ml inhibited C . neoformans growth by 82% . On native polyacrylamide gel electrophoresis (Nu-PAGE) the bound fraction migrated as a major and a minor band . Under the reducing conditions of sodium dodecyl sulfate (SDS)-PAGE the bound fraction yielded 4 prominent bands with MW ranging from 175 kDa to 45 kDa . Purification of the active Sephadex G-200 peak was achieved using an anion exchange column (DEAE-Sephacel) . Protein eluted with 0.1 M NaCl had strong anticryptococcal activity (12.5 micrograms/ml, 79% inhibition), which in SDS-PAGE migrated as a single band with an approximate MW of 85 kDA . This protein appears important in natural host resistance to C . neoformans and polymorphisms or deficiencies may have epidemiologic and diagnostic relevance.

Med Mycol, 1999 Aug, 37(4), 235 - 43
Scanning electron microscopy of encapsulated and non-encapsulated Cryptococcus neoformans and the effect of glucose on capsular polysaccharide release; Cleare W et al.; Cryptococcus neoformans has a polysaccharide capsule composed primarily of glucuronoxylomannan (GXM) . This study focuses on the morphology of both encapsulated and non-encapsulated organisms in the presence and absence of monoclonal antibodies (mAbs) and serum proteins, and the effect of glucose on capsular polysaccharide release . Examination of the encapsulated C . neoformans strains 24067 and 34873 by scanning electron microscopy (SEM) revealed globular cells covered with a loose fibrillar network which was most prominent during the early stationary phase . In the presence of GXM-binding mAbs or serum the capsule border became distinct and bud scars were evident in the fibrillar network . In contrast, SEM of strain 52817, a non-encapsulated mutant of 34873 revealed ovoid cells devoid of a fibrillar network with bud scars and small surface protrusions . mAb 2H1 bound to cells from strains 24067 and 34873 but not 52817 . No GXM was detected in supernatants of 52817 culture . For several strains, there was significantly more GXM in culture supernatants using high glucose media . In summary, our results indicate: i) SEM methods for studying capsular structure in C . neoformans; ii) no reactivity by GXM-binding mAb with a non-encapsulated strain; iii) the presence of distinctive bud scars in both encapsulated and non-encapsulated cells; and iv) dependence of GXM concentration on glucose concentration in culture media . The implications of these results are discussed.

Med Mycol, 1999 Jun, 37(3), 175 - 81
Melanin as a potential cryptococcal defence against microbicidal proteins; Doering TL et al.; Cryptococcus neoformans is an important fungal pathogen that synthesizes melanin when grown in the presence of phenolic substrates . The ability of C . neoformans to produce melanin is associated with virulence, but the specific role of melanin in the pathogenesis of infection is not clear . In this study the ability of C . neoformans melanin to bind proteins and protect against microbicidal peptides was investigated . Melanin was shown to bind a variety of proteins of fungal and mammalian origin . Melanin-protein interactions were dependent on the pH of the solution and on the amount of protein and melanin present . Melanized cells were less susceptible to killing by three microbicidal peptides: a defensin, a protegrin, and a magainin . Incubation of the microbicidal peptides with melanin particles, followed by removal of the melanin, reduced or abolished fungicidal activity, demonstrating interactions between peptides and melanin . The ability of melanin to bind proteins and to protect against microbicidal peptides suggests a protective function for melanin, whereby it sequesters microbicidal peptides and abrogates their activity.

Med Mycol, 1999 Jun, 37(3), 169 - 74
Fungemia during murine cryptococcosis sheds some light on pathophysiology; Lortholary O et al.; We studied fungemia over time in outbred mice infected with Cryptococcus neoformans and looked at its relationship with the intravenous (i.v.) inoculum size, tissue burden and survival . Fungemia was evaluated by culture of 10 microl of peripheral blood from living mice or by culture of buffy coats from sacrificed animals . For all inoculum sizes studied, fungemia could last several weeks after the i.v . inoculation . Individual susceptibility of outbred mice to cryptococcal infection was evidenced by variations in the course, duration and magnitude of fungemia and tissue localizations . These results suggest that the fungus can recirculate after the initial i.v . inoculation . Fungemia, assessed by culture of buffy coats, correlated with the extent of infection in the spleen, lung or brain (P<<0.001) on day 1 after inoculation but only with yeast burden in lung or spleen on day 8, thus demonstrating that brain reacts differently to C . neoformans infection than other organs . Comparison of blood culture techniques and examination of smears suggest that cryptococci might circulate within leucocytes . Finally, quantitative blood cultures may accurately assess the fungal load during experimental cryptococcosis.

Med Mycol, 1999 Jun, 37(3), 151 - 7
Production of nitric oxide by rat alveolar macrophages stimulated by Cryptococcus neoformans or Aspergillus fumigatus; Gross NT et al.; Cryptococcus neoformans and Aspergillus fumigatus are airborne fungi and the alveolar macrophages (AM) constitute a first line of host defence against both pathogens . We investigated the ability of rat AM to produce nitric oxide (NO) when challenged in vitro with C . neoformans, A . fumigatus conidia or inert silica particles alone and together with interferon gamma (IFN-Gamma) . The role of NO in the killing of C . neoformans as well as the relationship between phagocytosis of the yeast or A . fumigatus conidia and NO production by AM were studied . Both fungi, but not the inert particles induced a small but significant increase in NO production by AM . A synergistically enhanced NO production by AM was observed when each fungus, but not silica particles, were incubated together with IFN-Gamma . AM treated with IFN-Gamma and challenged with C . neoformans showed higher killing activity than untreated AM, a finding that correlated with increased NO production by AM . Both effects were reduced by an inhibitor of NO synthesis . Increased NO production by IFN-Gamma activated AM was found together with an increased accumulated attachment of A . fumigatus conidia and serum opsonized, but not unopsonized C . neoformans . The IFN-Gamma dependent increase in accumulated attachment of the fungi might be responsible for the synergistic effect of the fungi and IFN-Gamma on the NO production . Our data suggest that activated rat AM might efficiently use the antimicrobial nitric oxide system in the defence against these pathogens in the normal host.

Indian J Pathol Microbiol, 1999 Jan, 42(1), 63 - 8
Renal lesions associated with AIDS--an autopsy study; Lanjewar DN et al.; Kidneys from 55 cases (20 with HIV infection and 35 with AIDS) were studied by routine Haematoxylin and Eosin stains and special stains (PAS, PASM GMS, ZN, Mucicarmine and Congo red) to evaluate, glomerular, interstitial and vascular pathology . Twenty-four of the 35 (68.6%) cases of AIDS showed infective aetiology which included 17 cases (48.5%) of tuberculosis, 5 cases (14.4%) of fungal infection (3 cryptococcus neoformans and 2 candida species) and 2 cases (5.7%) of CMV infection . Other lesions noted were amyloidosis and tubular calcinosis . HIV associated nephropathy (HIVAN) was not detected in any of the cases . Intravenous drug abuse was not a risk factor in our cases which probably explains the absence of HIV associated nephropathy in the present study.

Zh Vopr Neirokhir Im N N Burdenko, 1999 Apr-Jun, (2), 33 - 6
{A cryptococcal granuloma of the midbrain}; Konovalov AN et al.; The paper describes the most infrequent case of cryptococcal granuloma of the midbrain in a HIV-negative female patient aged 41 years . The patient with midbrain lesion without signs of meningitis was found to have a bulky midbrain opercular formation that was regarded as a nodal glioma . The diagnosis of cryptococcal granuloma was established after removal of the formation (via occipito-transtentorial access with dissection of the lamina tecti) and pathomorphological examination . Microbiological studies verified the diagnosis . Despite the initiation of specific treatment with amphotericin B, the patient died on day 12 following surgery for cryptococcal meningoencephalitis.

Acta Paediatr, 1999 Jun, 88(6), 671 - 4
Fatal biphasic brainstem and spinal leptomeningitis with Cryptococcus neoformans in a non-immunocompromised child; Ruggieri M et al.; Cryptococcal meningitis is one of the most common life-threatening, invasive fungal infections of the central nervous system in patients with defective T-lymphocyte function . It is, however, unusual in children . We report on a non-immunocompromised 10-y-old boy without evidence of immunological abnormality who developed headache, vomiting, disturbances of consciousness and areflexia . Magnetic resonance imaging of the brain and the spinal cord revealed enlargement of the ventricles and high signal lesions in the leptomeninges at the level of the cerebral peduncles and the cervical and thoracic cord . Cerebrospinal fluid analysis was positive for Cryptococcus neoformans . He was treated with amphotericin B and was symptom-free within 1 wk . Despite an extended course of therapy his symptoms suddenly relapsed and he succumbed to the medical complications of cardiac and respiratory failure . Central nervous system appearances at postmortem were those of cryptococcal leptomeningitis.

AIDS, 1999 Jul 9, 13(10), 1249 - 53
Changes to AIDS dementia complex in the era of highly active antiretroviral therapy; Dore GJ et al.; OBJECTIVES: To determine the protective efficacy of highly active antiretroviral therapy (HAART) against AIDS dementia complex (ADC) relative to other initial AIDS-defining illnesses (ADIs), Australian AIDS notification data over recent years were examined . METHODS: All initial ADIs in Australia over the period 1992-1997 were included . Three initial ADI groups were established: ADC; other predominantly central nervous system (CNS) ADIs (toxoplasmosis and cryptococcosis); and non-CNS ADIs . For each ADI grouping, the proportion of total ADls, and median CD4 cell count in the pre-HAART era (1992-1995) were compared with the HAART era (1996 and 1997) . RESULTS: Initial ADls peaked in Australia in 1994 (n = 1049), with a gradual decline to 1996 (n = 722), and a marked decline in 1997 (n = 367) . ADC constituted 4.4% of initial ADIs over the period 1992-1995, but increased after the introduction of HAART to 6.0% in 1996 and 6.5% in 1997 (P = 0.02) . In contrast, the proportion of other CNS ADIs (1992-1995, 8.1%; 1996, 6.0%; 1997, 8.2%; P = 0.41) was stable over the period 1992-1997 . The median CD4 cell count at ADC diagnosis increased from 70/mm3 in 1992-1995 to 120/mm3 in 1996 and 170/mm3 in 1997 (P = 0.04) . Although the median CD4 cell count also increased significantly over this period for both other CNS ADIs (40-60/mm3; P = 0.02), and non-CNS ADIs (60-70/mm3; P = 0.02), the increase was small . CONCLUSION: A proportional increase in ADC compared with other ADIs and a marked increase in the median CD4 cell count at ADC diagnosis have occurred since the introduction of HAART in Australia . These changes suggest that HAART has a lesser impact on ADC than on other ADIs, with the poor CNS penetration of many antiretroviral agents a possible explanation.

Fungal Genet Biol, 1999 Jun, 27(1), 36 - 48
Molecular analysis of the Cryptococcus neoformans ADE2 gene, a selectable marker for transformation and gene disruption; Sudarshan S et al.; Cryptococcus neoformans is an important fungal pathogen of man . The incidence of cryptococcal disease has increased dramatically in patients immunocompromised because of HIV infection, organ transplantation, or treatment with cytotoxic chemotherapy or corticosteroids . This organism is an excellent model for molecular dissection of fungal pathogenesis and virulence factors . Here we report the nucleotide sequence of the C . neoformans serotype D genomic ADE2 gene, which encodes a phosphoribosylaminoimidazole carboxylase required for purine biosynthesis . Importantly, this version of the ADE2 gene has been used as the selectable marker for virtually all gene disruptions by transformation and homologous recombination in C . neoformans . We compare the nucleotide and amino acid sequences of the ADE2 gene and product to other highly related adenine biosynthetic genes and enzymes from other yeasts and fungi . We also describe a series of convenient ADE2 cassettes for gene disruption construct preparation . Finally, we have identified the ade2 mutations in strains M001 and M049, adenine auxotrophic mutants derived from the serotype A strain H99 . These mutant strains have served as recipients for targeted gene disruptions using the ADE2 gene . These studies should facilitate transformation and gene disruption approaches using the ADE2 selectable marker in this important human fungal pathogen .

J Clin Microbiol, 1999 Aug, 37(8), 2697 - 8
Comparative performance of the RapID Yeast Plus System and the API 20C AUX Clinical Yeast System; Smith MB et al.; The performance of the RapID Yeast Plus System (Innovative Diagnostic Systems, Norcross, Ga.), a 4-h micropanel using single-substrate enzymatic test reactions, was compared with that of the API 20C AUX Clinical Yeast System (bioMerieux Vitek, Hazelwood, Mo.), a 48- to 72-h carbohydrate assimilation panel . Two hundred twenty-five yeasts, yeast-like fungi, and algae, comprising 28 species and including 30 isolates of Cryptococcus neoformans, an important pathogen not tested in appreciable numbers in other comparisons, were tested by both methods . On initial testing, 196 (87.1%) and 215 (95.6%) isolates were correctly identified by the RapID and API systems, respectively . Upon repeat testing, the number of correctly identified isolates increased to 220 (97.8%) for the RapID system and 223 (99.1%) for the API system . Reducing the turbidity of the test inoculum to that of a no . 3 McFarland turbidity standard, which is below that recommended by the manufacturer, resulted in the correct identification of most of the isolates initially misidentified by the RapID system, including 10 of 30 C . neoformans isolates . Concordance between the RapID and API results after repeat testing was 97.3%.

J Antimicrob Chemother, 1999 Jun, 43(6), 817 - 24
Fluconazole, with or without dexamethasone for experimental cryptococcosis: impact of treatment timing; Lortholary O et al.; The time of initiation of fluconazole treatment with or without dexamethasone, and the impact on mycological outcome and drug pharmacokinetics were assessed in a murine model of disseminated cryptococcosis . Non-infected mice and mice with disseminated cryptococcosis were given saline, dexamethasone, or fluconazole +/- dexamethasone, 1 or 8 days after infection . Cfus were counted in tissues, and fluconazole concentrations were determined in plasma and tissues by HPLC and a bioassay . Despite fluconazole tissue and plasma concentrations which were above the minimal inhibitory concentration, the numbers of cfus in brain and lung tissues were reduced after early (P = 0.002 and 0.04, respectively), but not after late fluconazole treatment . The administration of dexamethasone did not have a deleterious effect on the number of cfus, fluconazole pharmacokinetics or antifungal activity . In conclusion, the size of the fungal burden influences the effective level of fluconazole activity in lung and brain . These results strongly suggest that potential antifungal agents should be studied following both early and late administration in experimental cryptococcosis.

Clin Exp Immunol, 1999 Jul, 117(1), 113 - 22
Interferon-gamma (IFN-gamma)-dependent protection and synthesis of chemoattractants for mononuclear leucocytes caused by IL-12 in the lungs of mice infected with Cryptococcus neoformans; Kawakami K et al.; We have recently demonstrated that IL-12 induced cellular inflammatory responses consisting mainly of accumulation of mononuclear leucocytes in the lungs of mice infected with Cryptococcus neoformans and protected mice against fulminant infection . We examined the involvement of endogenously synthesized IFN-gamma in such a response by investigating the effects of a neutralizing monoclonal antibody against this cytokine . The latter treatment completely abrogated the positive effects of IL-12 on survival of infected mice and prevented IL-12-induced elimination of microbials from the lungs . Histopathological examination showed that accumulation of mononuclear leucocytes in the infected lungs caused by IL-12 was clearly inhibited by anti-IFN-gamma MoAb . We also examined the local production of mononuclear cell-attracting chemokines such as monocyte chemotactic protein-1 (MCP-1), regulated upon activation, normal T cell expressed and secreted (RANTES), macrophage inflammatory protein-1alpha (MIP-1alpha), MIP-1beta and IFN-gamma-inducible protein 10 (IP-10) in the lungs using a reverse transcriptase-polymerase chain reaction (RT-PCR) method . We found that these chemokines were not synthesized in the infected lungs, while IL-12 treatment markedly induced their production . Interestingly, neutralizing anti-IFN-gamma MoAb strongly suppressed IL-12-induced production of these chemokines . Similar results were obtained with MCP-1 and MIP-1alpha when their synthesis was measured at the protein level using respective ELISA kits . Our results indicate that IFN-gamma plays a central role in the protective effects of IL-12 by inducing mononuclear leucocyte-attracting chemokines and cellular inflammatory responses.

AIDS, 1999 Apr 16, 13(6), 653 - 60
Antibody to capsular polysaccharide enhances the function of neutrophils from patients with AIDS against Cryptococcus neoformans; Monari C et al.; OBJECTIVE: To determine the contribution of anti-glucuronoxylomannan monoclonal antibody (MAb18B7) to the fungicidal capacity of polymorphonuclear leukocytes (PMNL) from HIV-infected patients towards Cryptococcus neoformans . DESIGN: Killing activity and superoxide anion generation were evaluated in the presence or absence of MAb18B7 in an in vitro system . METHODS: Killing activity was determined by colony forming unit inhibition assay . Superoxide generation was measured in the presence or absence of zymosan, C . neoformans, or Candida albicans . CD16, CD32, and CD64 molecules on PMNL were evaluated by cytofluorometric analysis . RESULTS: MAb18B7 strongly influenced the phagocytic and killing activities against encapsulated C . neoformans and consistently enhanced superoxide anion generation . Expression of CD16, and to a lesser extent CD64, on PMNL was required for MAb18B7-induced superoxide generation . By blocking CD16 and CD64 molecules with anti-CD16 and anti-CD64 MAb, a significant down-regulation of MAb18B7-induced fungicidal activity was observed . CONCLUSIONS: Our results demonstrate that MAb18B7 selectively enhances the killing mechanisms of PMNL from HIV-infected patients against encapsulated C . neoformans . The availability of CD16 and CD64 molecules on PMNL plays a critical role.

J Vasc Surg, 1999 Jul, 30(1), 189 - 92
Cryptococcal aortitis presenting as a ruptured mycotic abdominal aortic aneurysm; Deitch JS et al.; Mycotic processes occasionally complicate atherosclerotic aortic disease and usually require aggressive surgical therapy to control sepsis and prevent arterial rupture . Rarely, fungal organisms are responsible for primary infection of the abdominal aorta . We report the first case of Cryptococcal aortitis presenting as a ruptured abdominal aortic aneurysm . The surgical, pathologic, and microbiologic aspects of fungal aortitis are discussed.

J Nat Prod, 1999 Jun, 62(6), 824 - 8
Synthetic analogues of irlbacholine: a novel antifungal plant metabolite isolated from Irlbachia alata; Lu Q et al.; Irlbacholine and a series of related analogues were synthesized and their antifungal activities against Candida albicans, Cryptococcus neoformans, and Aspergillus fumigatus were assessed . The natural bisphosphocholine, irlbacholine, was the most potent compound, its 22-carbon chain length appearing to be optimal.

Mycoses, 1999 Apr, 42(1-2), 103 - 6
The green colour effect (GCE) of the killer strain Cryptococcus laurentii CBS 139 on Staib agar; Staib F; Attention is drawn to the observation that the type strain Cryptococcus laurentii CBS 139, producing killer toxins (mycocins) directed at Cr . neoformans var . gattii, causes a green colour effect (GCE) on Staib agar (Guizotia abyssinica creatinine agar) in combination with an intense assimilation of creatinine . Five (9.6%) out of 52 strains of Cr . laurentii of various origin, showed a GCE and intense creatinine assimilation . Further research must show if all Cr . laurentii strains, characterized by a GCE similar to that of the strain CBS 139, are also capable of producing killer toxins . For further ecological and epidemiological research on strains producing killer toxins directed against species of the genus Cryptococcus, it is proposed to use Staib agar as differential culture medium indicating both colour effects, i.e . the GCE and the brown colour effect (BCE).

Mycoses, 1999 Apr, 42(1-2), 17 - 9
Dosage adjustment of fluconazole during continuous renal replacement therapy (CAVH, CVVH, CAVHD, CVVHD); Pittrow L et al.; Continuous arterio-venous haemofiltration (CAVH), continuous veno-venous haemofiltration (CVVH), continuous arterio-venous haemodialysis (CAVHD) and continuous veno-venous haemodialysis (CVVHD) are increasingly used in patients with acute renal failure (ARF) . The elimination rate of fluconazole varies considerably depending on the procedure used . (In Germany, fluconazole is approved for the treatment of life-threatening fungal infections caused by Candida spp . and Cryptococcus neoformans at a dosage of up to 800 mg day-1.) The elimination rate of fluconazole by CVVHD depends on the combined dialysate/ultrafiltrate flow rate, but is much higher than achieved with CVVH and intermittent dialysis, with a fluconazole clearance in patients with CVVHD 2 l h-1 exceeding the values of healthy persons . To achieve therapeutic plasma levels during continuous renal replacement therapy, the same loading dose as in patients without renal failure should be administered, followed by a maintenance dose that is adjusted for anuric patients by multiplying by a factor that takes into account the extracorporeal elimination of the absorbed dose (CAVH, CVVH x 2.2, ultrafiltrate flow 0.5 l h-1; CAVHD, CVVHD x 3.8, combined dialysate/ultrafiltrate flow 1.5 l h-1) . Despite the broad therapeutic margin of fluconazole, drug monitoring is recommended to achieve therapeutic drug levels in life-threatening indications because there have been only a few investigations of this, all involving relatively low dosages (up to 200 mg day-1).

Mycoses, 1999 Apr, 42(1-2), 3 - 16
Administration of fluconazole in children below 1 year of age; Schwarze R et al.; For this review, 78 studies regarding the use of fluconazole in a total of 726 children below 1 year of age were evaluated . The range of fluconazole dosage was 2-50 mg kg-1 day-1, with 162 days being the maximum duration of treatment . According to current experience, fluconazole seems to be well tolerated and efficacious against systemic candidosis and candidaemia in children below 1 year of age, including neonates and very low-birthweight infants (VLBWIs) . The recommended daily dosage is 6 mg kg-1 . (In Germany, fluconazole is approved for children between 1 and 16 years in cases in which there is no therapeutic alternative for treatment of systemic infections caused by Candida spp . and Cryptococcus neoformans in a dosage of 3-6 mg kg-1 day-1 and for superficial Candida infections in a dosage of 1-2 mg kg-1 day-1.) In patients with impaired renal function, the daily dose should be reduced in accordance with the guidelines given for adults . In neonates during the first 2 weeks of life, this dosage should be administered only every 72 h . In weeks 2-4 of life, the same dose should be given every 48 h, following which daily dosing is appropriate . This posology is derived from the age-related pharmacokinetics of fluconazole, with a higher volume of distribution and a prolonged plasma elimination half-life, especially during the first month of life . Drug monitoring during treatment should be performed to ensure therapeutic plasma concentrations of fluconazole within a range between 4 and 20 micrograms ml-1 . The benefit of fluconazole should be investigated in prospective studies for treatment of systemic candidosis with administration of higher dosages as well as for early empiric therapy in VLBWIs.

J Neurol Sci, 1999 Mar 15, 164(1), 72 - 5
Rapid diagnosis of cryptococcal meningitis by microscopic examination of centrifuged cerebrospinal fluid sediment; Sato Y et al.; The classic India ink test is positive in only half of cryptococcal meningitis cases, and reliable, rapid cryptococcal antigen (CRAG) testing requires technical expertise and facilities not always available . We therefore examined cerebrospinal fluid (CSF) sediment using May-Giemsa, periodic acid-Schiff, and Gram stains in 16 patients with cryptococcal meningitis . The India ink test was positive in seven patients (44%), while microscopic examination of sediment revealed cryptococci in 13 (81%); in six of these 13 the India ink test was negative . Both methods failed to detect the pathogen in the remaining three patients . CRAG testing in CSF was negative in two patients (one with acquired immunodeficiency syndrome, one with diabetes mellitus) whose India ink test also was negative while cryptococci were identified in their CSF sediment . No false positives occurred with CSF May-Giemsa staining in 27 cases of aseptic meningitis with negative cultures for Cryptococcus . In all, microscopic examination of centrifuged and stained CSF sediment proved more sensitive for rapid diagnosis of cryptococcal meningitis than the India ink method, and in two of our patients cryptococci were seen in centrifuged CSF sediment despite negative CRAG and India ink tests.

Arch Pathol Lab Med, 1999 Jun, 123(6), 519 - 23
Neuropathology of human immunodeficiency virus 1 infection . Significance of studying in forensic autopsy cases at Dar es Salaam, Tanzania; Kibayashi K et al.; OBJECTIVE: In sub-Saharan Africa, only a few studies of neurologic complications of human immunodeficiency virus 1 (HIV-1) infection have been done . The authors studied neuropathology of HIV-1 infection in Tanzania . DESIGN: Forensic autopsy study at Dar es Salaam, Tanzania . SETTING: A joint research project between Dar es Salaam, Tanzania, and Kumamoto, Japan . PATIENTS: Thirty patients with risk factors for HIV-1 infection . MAIN OUTCOME MEASURES: Human immunodeficiency virus 1 infection was evaluated by HIV-1 antibody test on postmortem serum samples . The brains of HIV-1-infected persons were studied histopathologically . RESULTS: Infection with HIV-1 was identified on postmortem serum samples in 10 of 30 forensic autopsy cases . Neuropathologic changes of the brain were observed in 8 of the 10 HIV-1-infected persons; these changes consisted of lymphocytic meningitis, bacterial meningoencephalitis, cryptococcal meningoencephalitis, tuberculous meningitis with brain abscesses, and intracerebral hemorrhage . CONCLUSIONS: Because none of the persons studied was suspected to have had brain diseases before autopsy, the results suggest that brain diseases of HIV-1-infected patients are likely to go unrecognized in Tanzania . In addition, the high incidence of neuropathologic findings in HIV-1-infected persons indicates that HIV-1-related brain diseases are common in Tanzania, as they are in developed countries . Further forensic autopsy study will determine the range and prevalence of brain complications and have immediate impact on the management of HIV-1-infected patients in Tanzania and other developing countries.

Rev Soc Bras Med Trop, 1999 May-Jun, 32(3), 255 - 61
{Usefulness of routine investigation of fungal infections through bronchoscopy in HIV-infected and non-HIV-infected patients in a general hospital, reference to AIDS}; Lazzarini-de-Oliveira LC et al.; The diagnostic yields and the spectrum of pulmonary fungal-infection obtained in samples collected by fiberoptic bronchoscopy from HIV-positive and HIV-negative patients were evaluated from 1990 to 1995 . A total of 1943 bronchoscopies were performed during this period, 47% in the HIV-positive group and 53% in the HIV-negative group . Of 908 HIV-positive patients, 38 (4%) had a fungus isolated from the pulmonary sample whereas of 1035 HIV-negative patients, only 4 (0.2%) had a fungus isolated . Histoplasmosis and Cryptococcosis were more frequently found in HIV-positive than in HIV-negative patients (p < 0.001) . Paracoccidioides brasiliensis was found in only 3 patients, all of them immunocompetent . The study demonstrated that, despite the low yields, the HIV-positive group may benefit from routine screening for fungal elements in specimens obtained by fiberoptic bronchoscopy.

J Neuroophthalmol, 1999 Jun, 19(2), 131 - 5
Wall-eyed bilateral internuclear ophthalmoplegia in central nervous system cryptococcosis; Fay PM et al.; Only one case of wall-eyed bilateral internuclear ophthalmoplegia (WEBINO) has been described in central nervous system cryptococcosis . The disorder was initially unilateral, then became bilateral with skew deviation and vertical upgaze deficit . We report a case of WEBINO in central nervous system cryptococcosis in a patient with acquired immune deficiency syndrome . Magnetic resonance imaging revealed high signal on T2 images in the right midbrain, left frontal vertex, left splenium, and cerebellum . With treatment, the internuclear ophthalmoplegia improved; however, the convergence insufficiency remained . Disruption of input from cortical supranuclear locations or the region of the rostral interstitial nucleus of the medial longitudinal fasciculus has been proposed as a mechanism in the absence of convergence . This correlates in our patient with the lesions seen on magnetic resonance images.

J Neuroophthalmol, 1999 Jun, 19(2), 118 - 9
Bilateral trochlear nerve palsy associated with cryptococcal meningitis in human immunodeficiency virus infection; Sadun F et al.; This is the report of a case of bilateral trochlear nerve palsy secondary to cryptococcal meningitis in a 34-year-old woman with acquired immune deficiency syndrome . Based on clinical and neuroradiologic findings, it is concluded that in the present case, a postinflammatory shrinking of the arachnoid has stretched the fourth cranial nerves at their point of emergence from the dorsal surface of the brainstem.

Neurologia, 1999 May, 14(5), 218 - 23
{Meningitis by Cryptococcus neoformans in patients with HIV infection}; Martinez Fernandez EM et al.; OBJECTIVE: To review a serie of patients with cryptococcal meningitis and immunodeficiency syndrome (AIDS) treated in our hospital in the last two years . PATIENTS AND METHODS: Retrospective study of 25 patients infected with the human immunodeficiency virus (HIV) and affected by Cryptococcus neoformans meningitis . The factors analysed were epidemiological data, clinical manifestations, biochemical and microbiological characteristics of cerebrospinal fluid (CSF), radiological abnormalities, treatment, adverse reactions and outcomes . RESULTS: Eighty-four percent of patients had less than 200 CD4/microliter . Cryptococcal infection was the AIDS defining illness in 24% of cases . Patients typically presented with neurologic symptoms such as: headache (88%), fever (68%) and somnolence (68%); 20% presented seizures and 28% focal deficits . There were no CSF biochemical alterations in 25% of them . CSF culture and indian ink stain were positive in 76% . CSF cryptococcal antigen test was positive in 68% of the cases . TC showed abnormalities in 48% . CSF of all patients treated with amphotericin B (AB) plus flucytosine (5FC) whose CSF culture was monitored became negative in the first two weeks, meanwhile those treated only with AB or fluconazol had negative control culture in 60% and 50% respectively . Six patients died within the initial 10 weeks . Death was due to bacterian sepsis in 3 patients and high intracranial pressure was the cause in 2 cases . One happened before treatment was administered . CONCLUSIONS: It's essential to consider the possibility of cryptococcal meningitis in patients infected with HIV and any compatible symptom regardless of CSF biochemical results and immunodepression level (CD4) . Although our study was non randomized and so we can't propose a therapeutical schedule based on it, we can say that patients treated with AB plus 5FC showed an earlier conversion from positive to negative CSF cultures without more adverse reactions.

J Med Chem, 1999 Jun 17, 42(12), 2112 - 24
Phthalein derivatives as a new tool for selectivity in thymidylate synthase inhibition; Costi PM et al.; A new set of phthalein derivatives stemming from the lead compound, phenolphthalein, were designed to specifically complement structural features of a bacterial form of thymidylate synthase (Lactobacillus casei, LcTS) versus the human TS (hTS) enzyme . The new compounds were screened for their activity and their specificity against TS enzymes from different species, namely, L . casei (LcTS), Pneumocystis carinii (PcTS), Cryptococcus neoformans (CnTS), and human thymidylate synthase (hTS) . Apparent inhibition constants (Ki) for all the compounds against LcTS were determined, and inhibition factors (IF, ratio between the initial rates of the enzymatic reaction in the presence and absence of each inhibitor) against each of the four TS species were measured . A strong correlation was found between the two activity parameters, IF and Ki, and therefore the simpler IF was used as a screening factor in order to accelerate biological evaluation . Compounds 5b, 5c, 5ba, and 6bc showed substantial inhibition of LcTS while remaining largely inactive against hTS, illustrating for the first time remarkable species specificity among TSs . Due to sequence homology between the enzymes, several compounds also showed high activity and specificity for CnTS . In particular, 3-hydroxy-3-(3-chloro-4-hydroxyphenyl)-6-nitro-1H, 3H-naphtho{1,8-c,d}pyran-1-one (6bc) showed an IF < 0.04 for CnTS (Ki = 0.45 microM) while remaining inactive in the hTS assay at the maximum solubility concentration of the compound (200 microM) . In cell culture assays most of the compounds were found to be noncytotoxic to human cell lines but were cytotoxic against several species of Gram-positive bacteria . These results are consistent with the enzymatic assays . Intriguingly, several compounds also had selective activity against Cr . neoformans in cell culture assay . In general, the most active and selective compounds against the Gram-positive bacteria were those designed and found in the enzyme assay to be specific for LcTS versus hTS . The original lead compound was least selective against most of the cell lines tested . To our knowledge these compounds are the first TS inhibitors selective for bacterial TS with respect to hTS.

Infect Immun, 1999 Jul, 67(7), 3601 - 9
Differential regulation of immune responses by highly and weakly virulent Cryptococcus neoformans isolates; Blackstock R et al.; Early inflammatory responses, delayed-type hypersensitivity (DTH) responses, and cytokine profiles were studied in mice infected by the pulmonary route with either a highly virulent isolate (NU-2) or a weakly virulent isolate (184A) of Cryptococcus neoformans . After infection, NU-2 remained in the lungs and the capsule became more pronounced during the first 24 h, whereas 184A induced an immediate inflammatory reaction and was rapidly cleared from the lungs . Cryptococcal antigen (GXM) appeared in sera early after infection with NU-2 and increased over the entire observation period . There was no detectable GXM in sera from 184A-infected mice . Both C . neoformans isolates induced anticryptococcal cell-mediated immune responses, but the responses had different profiles . DTH in NU-2-infected mice appeared at day 15 after infection and waned by day 21, whereas DTH in 184A-infected mice was present by day 5 and continued to increase . T helper 1 (Th1) cytokines (interleukin 2 {IL-2} and gamma interferon) were made by spleen cells early after infection with either isolate . NU-2-infected mice lost their ability to produce these cytokines, but 184A-infected mice retained it . IL-4, a Th2 cytokine, was not detected in infected mice . The regulatory cytokine IL-10 was made by spleen cells early but not later after infection with the highly virulent isolate and was not produced by spleen cells from 184A-infected mice . IL-10-deficient mice survived an NU-2 infection significantly longer than wild-type mice, suggesting that IL-10 is important in down-regulating the protective immune response . The induction of anergy appears to be responsible for the inability of NU-2-infected mice to control a C . neoformans infection.

AJNR Am J Neuroradiol, 1999 May, 20(5), 867 - 75
Magnetization transfer MR imaging in CNS tuberculosis; Gupta RK et al.; BACKGROUND AND PURPOSE: CNS tuberculosis may simulate other granulomas and meningitis on MR images . The purpose of this study was to improve the characterization of lesions in CNS tuberculosis and to assess the disease load using magnetization transfer (MT) imaging . METHODS: A total of 107 tuberculomas in seven patients with or without meningitis and 15 patients with tuberculosis meningitis alone were studied . Fifteen patients with cysticercus granulomas with T2 hypointensity, five patients each with viral and pyogenic meningitis, and two patients with cryptococcal meningitis were also studied . The MT ratios were calculated from tuberculomas, cysticercus granulomas, and thickened meninges in tuberculous, viral, pyogenic, and cryptococcal meningitis and were compared within each pathologic group and with the MT ratio of different regions of normal brain parenchyma . Detectability of lesions on T1-weighted MT spin-echo (SE) images was compared with that on conventional SE and postcontrast MT-SE images . RESULTS: Thickened meninges appeared hyperintense relative to surrounding brain parenchyma in the basal and supratentorial cisterns on precontrast MT-SE images in all 18 patients with tuberculosis meningitis . These meninges were not seen or were barely visible on conventional SE images, and enhanced on postcontrast MT-SE images . The MT ratio from the thickened meninges of tuberculous meningitis was significantly lower than that from the meninges in cryptococcal and pyogenic disease and significantly higher than the meninges in viral meningoencephalitis . The MT ratio from T2 visible and invisible tuberculomas appeared to be significantly lower than that of normal white matter . The MT ratio of T2 hypointense cysticercus granuloma was significantly higher than that of T2 hypointense tuberculoma . CONCLUSION: Precontrast MT-SE imaging helps to better assess the disease load in CNS tuberculosis by improving the detectability of the lesions . With the use of MT ratios, it may be possible to differentiate tuberculosis from similar-appearing infective lesions on MR images.

Prikl Biokhim Mikrobiol, 1999 Mar-Apr, 35(2), 190 - 3
{Activity of exoglycans as sorbents of heavy metal ions}; Elinov NP et al.; The ability of yeast exoglycans (from Cryptococcus laurentii, Cr . luteolus, and Sporobolomyces albo-rubescens) to absorb copper and lead ions has been studied . The sorption isotherms have been obtained, which indicates that the mechanism of interactions in the system polysaccharide-metal is complex . For the majority of the glycans, the absorption selectivity was considerably higher in the case of copper, as compared to lead . The observed discrepancy in the effects of pH and temperature on the absorption indicates that the bonds mediating ion absorption by glycans are of different nature . In conclusion, yeast exoglycans are efficient sorbents of copper and lead ions, and lutelan is the most active exoglycan in this respect.

J Neurol, 1999 Apr, 246(4), 237 - 43
Opportunistic infections of the central nervous system during HIV-1 infection (emphasis on cytomegalovirus disease); Roullet E; Toxoplasma encephalitis, cryptococcal meningitis, progressive multifocal leukoencephalopathy (PML), and cytomegalovirus (CMV) encephalitis are the most common opportunistic infections of the central nervous system (CNS) in HIV-infected patients . They occur at variable degrees of immunosuppression, and their diagnosis is based on a systematic evaluation with includes, in a definite order, ongoing prophylactic therapies, extraneurological signs, neuroimaging and CSF studies, and an anti-Toxoplasma therapeutic trial . Concurrent neurological HIV-CNS disease (such as the AIDS dementia complex) is frequent . The development of reliable molecular biology techniques such as the polymerase chain reaction and their application to the CSF have made the diagnosis of virus-related opportunistic infections much easier and has limited the need for cerebral biopsy . The incidence of opportunistic infections has decreased since the introduction of recent antiretroviral therapeutic strategies.

FEMS Microbiol Lett, 1999 Jun 1, 175(1), 87 - 94
Differential effect of Cryptococcus neoformans on the production of IL-12p40 and IL-10 by murine macrophages stimulated with lipopolysaccharide and gamma interferon; Kawakami K et al.; In the present study, we examined the in vitro effect of Cryptococcus neoformans on the production of interleukin-12 (IL-12) and IL-10 by murine macrophages . At a dose of 1 x 10(5), 1 x 10(6) or 1 x 10(7) ml-1, a highly virulent strain of C . neoformans (strain YC-11) suppressed the production of IL-12p40 by a murine macrophage cell line, J774.1 stimulated with lipopolysaccharide (LPS) and interferon (IFN)-gamma, while the production of IL-10 was not inhibited, but rather slightly augmented . The suppression of IL-12p40 production did not change by neutralizing anti-IL-10 mAb . A direct contact of C . neoformans with macrophages was largely involved in this inhibitory effect, since placement of a 0.45 micron pore membrane between the organism and macrophages prevented such effect . On the other hand, the culture supernatant of YC-11 did not inhibit macrophage IL-12p40 production when used at a lower dose, which contained an equivalent amount of capsular polysaccharide to that in the supernatant of YC-11 cultured at 1 x 10(5) or 1 x 10(6) ml-1, although it showed a small suppression at higher doses . Our results suggest that C . neoformans may suppress the induction of Th1 responses by inhibiting macrophage IL-12 production predominantly through a direct contact-dependent mechanism and to a lesser extent by a certain soluble factor(s) released from this microorganism.

Chem Pharm Bull (Tokyo), 1999 May, 47(5), 643 - 6
Regiospecific synthesis of pyrido{3,4-b}- and pyrido{4,3-b}carbazole-5,11-dione derivatives . Evaluation of their in vitro antifungal or antiprotozoological activities; Poumaroux A et al.; Hetero Diels-Alder reactions between 2- or 3-bromocarbazolequinones 1a or 1b and azadiene 5 afford regiospecifically pyrido{3,4-b}- and pyrido{4,3-b}carbazole-3,5,11-triones 6a and 6b . The regiochemistry of the cycloadditions is controlled by the position of the bromine atom at C-2 or C-3 of the bromoquinone . The corresponding N- and O-methyl derivatives 7 and 8 are prepared . Structural assignment of the regioisomers is made by 1H-NMR nuclear Overhauser effect difference experiments performed on a diacetoxy derivative of pyrido{4,3-b}carbazole 9b . The in vitro antifungal and antiprotozoological activities of some prepared derivatives have been evaluated against Candida albicans, Candida krusei, Cryptococcus neoformans and Trichomonas vaginalis . None of the tested compounds have shown significant activity towards the yeasts or protozoa.

Clin Electroencephalogr, 1999 Apr, 30(2), 35 - 8
Discrimination between viral and nonviral meningitis by visually analyzed and quantitative electroencephalography; Bartel P et al.; A prospective study was conducted to assess the ability of the visually analyzed electroencephalogram (VEEG), the quantitative EEG (QEEG) and the Glasgow Coma Scale (GCS) to discriminate between patients with viral and nonviral meningitis . The 55 subjects, aged 14-75 years, fell into one of the following categories: viral (n = 12), bacterial (n = 19), tuberculous (n = 16) or cryptococcal (n = 8) meningitis . EEG recordings and Glasgow Coma Scale (GCS) scores were obtained within 48 hours of admission to hospital . The sensitivity of the VEEG and QEEG for the prediction of patients with nonviral meningitis (true positives in this context) attained reasonably high values of 70% and 80%, respectively . In contrast, the sensitivity of the GCS was only 38% . Each of the three tests achieved high degrees of consistency in this regard with positive predictive values of 94% or better . The specificity for each of the three tests was high, 100% for the VEEG and the GCS and 82% for the QEEG indicating a high probability for the correct prediction of viral meningitis (true negatives) . The consistency of this prediction was, however, poor due to negative predictive values of only 53% for the QEEG, 48% for the VEEG and 32% for the GCS . The QEEG results did not reveal any obvious advantages over the VEEG . Rather the assessment of the occurrence of particular VEEG abnormalities showed that patients with delta abnormalities had a very high probability of nonviral meningitis . At the other end of the spectrum, all normal VEEGs occurred in viral meningitis . In important respects the predictive ability of the EEG was superior to that of the GCS . While there was statistically significant agreement between the VEEG and GCS, the degree of agreement was poor . This study indicates that the EEG is a valuable and probably underestimated test in the acute phase of meningitis and provides complementary information to the GCS.

Diagn Microbiol Infect Dis, 1999 Jun, 34(2), 111 - 4
Use of cerebrospinal fluid shunt for the management of elevated intracranial pressure in a patient with active AIDS-related cryptococcal meningitis; Mylonakis E et al.; Persistently elevated intracranial pressure (ICP) is one of the most accurate predictors of a poor prognosis in patients with AIDS-related cryptococcal meningitis . We present a severe case of persistent cryptococcal meningitis in a patient with advanced AIDS, complicated by elevation of ICP . A ventriculoperitoneal shunt was placed that successfully lowered the ICP and alleviated the associated symptoms . The elevated ICP secondary to AIDS-related cryptococcal meningitis should be treated aggressively . Despite the risk of shunt complications, cerebrospinal fluid shunts can be considered in these patients if they do not respond to other treatment.

J Infect Dis, 1999 Jul, 180(1), 87 - 92
Fever and human immunodeficiency virus infection as sentinels for emerging mycobacterial and fungal bloodstream infections in hospitalized patients >/=15 years old, Bangkok; Archibald LK et al.; To determine the etiology of bloodstream infections (BSIs) in hospitalized patients >/=15 years old in Thailand, prospectively enrolled, consecutive febrile (>/=38 degrees C) patients were admitted to one hospital during February-April 1997 . After a patient history was taken and a physical examination was performed, blood was obtained for comprehensive culture and human immunodeficiency virus (HIV) testing . Of 246 study patients, 119 (48%) had BSIs, and 182 (74%) were infected with HIV . The 2 most common pathogens were Cryptococcus neoformans and Mycobacterium tuberculosis (30 and 27 patients, respectively) . HIV-positive patients were more likely than HIV-negative patients to have mycobacteremia (57/182 vs . 0/64, P< . 0001), fungemia (38/182 vs . 2/64, P<.001), or polymicrobial BSIs (19/182 vs . 0/64, P<.002) . Clinical predictors of BSIs included HIV infection, chronic diarrhea, lymphadenopathy, or splenomegaly . Mortality was higher among patients with than those without BSIs (P< . 001) . Cohort-based microbiologic studies are critically important to diagnose emerging pathogens and to develop algorithms for empirical treatment of BSIs in developing countries.

Diagn Cytopathol, 1999 Jun, 20(6), 385 - 6
Cryptococcosis of thoracic vertebra simulating tuberculosis: diagnosis by fine-needle aspiration biopsy cytology--a case report; Jain M et al.; A rare case of cryptococcosis of sixth thoracic vertebra (T6) along with pulmonary involvement in an old diabetic patient is presented . The infection resulted in lytic lesion of T6 vertebra and girdle pain . A computerized tomographic (CT) guided fine-needle aspiration biopsy (FNAB) cytology was performed, which showed encapsulated fungal spores of Cryptococcus neoformans with granulomatous reaction, later confirmed by fungal culture.

Mycopathologia, 1998-99, 143(3), 131 - 4
Recovery of Cryptococcus neoformans from the nasopharynx of AIDS patients; Sukroongreung S et al.; Nasopharyngeal swabbings, obtained from AIDS patients, were plated onto Niger seed agar containing antibiotics Cryptococcus neoformans was isolated from 35 out of 84 patients (41.7%) diagnosed as primary cryptococcal cases before antifungal administration, and 8 out of 86 (9.3%) cryptococcosis patients on antifungal therapy . The fungus could not be isolated from any of 447 samples from 194 AIDS patients not diagnosed with cryptococcosis . These findings are novel in that the presence of C . neoformans in AIDS patients at this site has never been looked at previously.

Mycopathologia, 1998-99, 143(3), 121 - 5
Human defenses against Cryptococcus neoformans: an update; Brummer E; Cryptococcus neoformans var . neoformans is an opportunistic fungal pathogen, especially in AIDS patients, and is found world-wide . On the other hand, Cryptococcus neoformans var . gatti (CN-g) is restricted to an association with two species of Eucalyptus trees . Alveolar macrophages (AM) constitute the first line of defense to Cryptococcus neoformans and offers some resistance . The inflammatory response to Cryptococcus neoformans with an influx of neutrophils and monocytes affords a second line of defense . Secretion of proinflammatory monokines by human AM is now being defined . The inflammatory phagocytes are efficient in killing Cryptococcus neoformans and offer strong resistance . T and B cell responses to infection, a third line of defense, results in production of lymphokines (IFNg, etc.) and specific antibodies . Enhancement of lymphocyte responses by IL-12 and IL-18 to Cryptococcus neoformans infection appears to be critical . Susceptibility of AIDS patients to Cryptococcus neoformans is associated with low CD4+ T cell counts and likely reduced efficacy of the second line of defense.

Gene, 1999 May 31, 232(2), 155 - 63
Characterization of the glyceraldehyde-3-phosphate dehydrogenase gene {correction of glyceraldehyde-3-phosphate gene} and the use of its promoter for heterologous expression in Cryptococcus neoformans, a human pathogen; Varma A et al.; The GPD gene encoding glyceraldehyde-3-phosphate dehydrogenase was isolated from Cryptococcus neoformans, a heterobasidiomycetous yeast that is pathogenic to humans . The gene contains 11 introns, differing from the conserved intron positions found in the GPD genes of Basidiomycetes . The predicted amino-acid sequence of this gene is extremely similar to that reported from GPD proteins of other basidiomycetes . The promoter region of the C . neoformans GPD gene was similar to those of other basidiomycetes . Plasmid constructs containing up to 1600 base pairs upstream of the native GPD open reading frame were used to express either the native URA5 gene in a ura5 mutant or the heterologous hphI gene (a bacterial gene that confers resistance to the aminoglycoside hygromycin) in a wild-type strain of C . neoformans . Transformation frequencies resulting from the plasmid-borne Gpdp::URA5 gene were at levels similar to those of the native URA5, which suggested that all the sequences necessary for proper expression were present . Transformation frequencies using the Gpdp::hphI gene constructs were poor . However, addition of DNA sequences flanking the 3'-end of an native C . neoformans gene significantly improved the transformation frequencies resulting from the expression of the heterologous hphI gene.

Clin Neurol Neurosurg, 1999 Mar, 101(1), 23 - 5
Midbrain infarction: a rare presentation of cryptococcal meningitis; Kalita J et al.; A 20-year-old farmer who had headache and fever for 1 month, suddenly developed left hemiplegia, tremor in left arm and titubation followed by deep coma . Cranial CT scan revealed an infarction in right crus of midbrain . His CSF revealed 66 mg/dl protein, 10 lymphocytes/mm3, and 70 mg/dl glucose . CSF was positive for cryptococcal antigen . He improved following i.v . amphotericin 0.5 mg/kg and fluconazole 200 mg daily, continued for 6 and 12 weeks respectively . Infarctions though rare in cryptococcal meningitis should be considered in patients with chronic meningitis with vasculitis.

Rev Cubana Med Trop, 1998, 50(3), 207 - 8
{The evaluation of the specific type of capsular polysaccharide of Cryptococcus neoformans as an immunogen and positive-control antigen}; Torano Peraza GT et al.; The specific type of capsular polysaccharide of an autochthonous strain of Cryptococcus neoformans var . neoformans was obtained by using the method of selective precipitation with hexadimethrine bromide . The capsular polysaccharide was matched to lamb's erythrocytes and it was used as an immunogen in rabbits . Antibody titres of up to 1:32 were attained . Doble serial dilutions of the capsular polysaccharide were evaluated as positive control antigen by contraimmunoelectrophoresis, latex and ELISA techniques, showing biological activity.

Rev Cubana Med Trop, 1998, 50(2), 167 - 9
{The identification of Cryptococcus neoformans var . neoformans in Cuban clinical isolates}; Fernandez Andreu CM et al.; 50 strains of Cryptococcus neoformans of clinical origin were studied by using the canavanine-glycine-bromothymol blue (CGB) medium . 56% of the strains were isolated from AIDS patients, and 16% belonged to patients with kidney transplantation . 90% of the samples were obtained from the samples of cerebrospinal fluid, which corresponded to the classical form of presentation of cryptococcosis . All the strains were identified as C . neoformans var . neoformans, coinciding with previous reports made in Cuba . Knowing the varieties of C . neoformans, inferences can be drawn on the epidemiology, clinics and response to the treatment of cryptococcosis.

Antimicrob Agents Chemother, 1999 Jun, 43(6), 1401 - 5
In vitro antifungal activity of nikkomycin Z in combination with fluconazole or itraconazole; Li RK et al.; Nikkomycins are nucleoside-peptide antibiotics produced by Streptomyces species with antifungal activities through the inhibition of chitin synthesis . We investigated the antifungal activities of nikkomycin Z alone and in combination with fluconazole and itraconazole . Checkerboard synergy studies were carried out by a macrobroth dilution procedure with RPMI 1640 medium at pH 6.0 . At least 10 strains of the following fungi were tested: Candida albicans, other Candida spp., Cryptococcus neoformans, Coccidioides immitis, Aspergillus spp., and dematiacious fungi (including Exophiala jeanselmei, Exophiala spinifera, Bipolaris spicifera, Wangiella dermatitidis, Ochroconis humicola, Phaeoannellomyces werneckii, and Cladophialophora bantiana), and 2 strains each of Fusarium, Scedosporium, Paecilomyces, Penicillium, and Trichoderma spp . A total of 110 isolates were examined . Inocula of fungal elements were standardized by hemacytometer counting or spectrophotometrically . MICs and minimum lethal concentrations (MLCs) were determined visually by comparison of growth in drug-treated tubes with growth in drug-free control tubes . Additive and synergistic interactions between nikkomycin and either fluconazole or itraconazole were observed against C . albicans, Candida parapsilosis, Cryptococcus neoformans, and Coccidioides immitis . Marked synergism was also observed between nikkomycin and itraconazole against Aspergillus fumigatus and Aspergillus flavus . No antagonistic interaction between the drugs was observed with any of the strains tested.

J Antibiot (Tokyo), 1999 Mar, 52(3), 305 - 10
Mulundocandin, an echinocandin-like lipopeptide antifungal agent: biological activities in vitro; Hawser S et al.; Mulundocandin (MCN) is an antifungal lipopeptide which belongs to the echinocandin class of antimycotic agents . MCN exhibited good in vitro activity against Candida albicans and C . glabrata isolates with MIC ranges of 0.5-4.0 microg/ml and 2.0-4.0 microg/ml, respectively . MCN also exhibited some activity against C . tropicalis isolates (MIC range 1.0-8.0 microg/ml) . However, MCN was poorly active against other non-albicans isolates and was inactive against Cryptococcus neoformans, Aspergillus species and Trichophyton . MCN appeared to exert its antifungal activity through preferential inhibition of germ tube formation (MIC-HY 0.015-0.03 microg/ml) and was typically less active on the yeast form (MIC 0.5-4.0 microg/ml) . In kill-curve experiments 99.9% reductions in cell viability were observed following 8 hours exposure to MCN at 4 x MIC and 8 x MIC and after 5 hours exposure to 16 x MIC.

J Nat Prod, 1999 May, 62(5), 674 - 7
Antifungal jujubogenin saponins from Colubrina retusa; Li XC et al.; Antifungal assay-guided isolation of the 95% ethanol extract of the stems of Colubrina retusa yielded jujubogenin 3-O-alpha-L-arabinofuranosyl(1-->2)-{beta-D-glucopyranosyl (1-->3)}-alpha-L-arabinopyranoside (1), which showed modest growth-inhibitory effects against Candida albicans, Cryptococcus neoformans, and Aspergillus fumigatus (MICs, 50 microg/mL) . In addition, two new minor saponins, jujubogenin 3-O-alpha-L-arabinofuranosyl(1-->2)-{2-O-(trans, cis)p-coumaroyl-beta-D-glucopyranosyl(1-->3)}-alpha-L-arabinopy ranosi de (2), and jujubogenin 3-O-(5-O-malonyl)-alpha-L-arabinofuranosyl (1-->2)-{beta-D-glucopyranosyl(1-->3)}-alpha-L-arabinopyranoside (3), were obtained . Saponin 2 was marginally active against only C . neoformans, with a MIC of 50 microg/mL, while 3 was inactive . NMR spectroscopy was used extensively for the structure determination of these compounds . The previously reported ambiguity of the NMR assignments of jujubogenin saponins for carbons -26 to -29 was clarified by a comprehensive analysis of the NMR spectra of 1.

Infect Immun, 1999 Jun, 67(6), 3096 - 107
In vitro and in vivo stability of a Cryptococcus neoformans {corrected} glucuronoxylomannan epitope that elicits protective antibodies; Cleare W et al.; The monoclonal antibody (MAb) 2H1 defines an epitope in Cryptococcus neoformans capsular glucuronoxylomannan (GXM) that can elicit protective antibodies . In murine models of cryptococcosis, MAb 2H1 administration prolongs survival and reduces fungal burden but seldom clears the infection . The mechanism by which C . neoformans persists and escape antibody-mediated clearance is not understood . One possibility is that variants that do not bind MAb 2H1 emerge in the course of infection . Using an agglutination-sedimentation protocol, we recovered a variant of strain 24067 that did not agglutinate, could not be serotyped, and had marked reduction in GXM O-acetyl groups . Binding of MAb 2H1 to 24067 variant cells produced a different immunofluorescence pattern and lower fluorescence intensity relative to the parent 24067 cells . Addition of MAb 2H1 to 24067 variant cells had no effect on cell charge . Phagocytic assays demonstrated that MAb 2H1 was not an effective opsonin for the 24067 variant . The 24067 variant was less virulent than the 24067 parent strain in mice, and MAb 2H1 administration did not prolong survival in animals infected with the variant strain . To investigate whether variants which do not bind MAb 2H1 are selected in experimental infection, three C . neoformans strains were serially passaged in mice given either MAb 2H1 or no antibody . Analysis of passaged isolates by agglutination assay, flow cytometry, and indirect immunofluorescence revealed changes in MAb 2H1 epitope expression but no clear trend with regards to gain or loss of MAb 2H1 epitope . C . neoformans variants with reduced MAb 2H1 epitope content can be isolated in vitro, but persistence of infection in mice given MAb 2H1 does not appear to be a result of selection of escape variants that lack the MAb 2H1 epitope.

AIDS Res Hum Retroviruses, 1999 May 1, 15(7), 625 - 31
Risk factors for cryptococcal meningitis in HIV-infected patients; Oursler KA et al.; To identify the risk factors for cryptococcal meningitis in patients with HIV disease we conducted a nested case-control study of 37 incident cases of cryptococcal meningitis and 74 controls, identified from a cohort of more than 2000 HIV-infected patients . Conditional logistic regression was used to study demographic and AIDS-related variables in addition to fluconazole and steroid use . No difference in demographic variables, HIV risk factors, or stage of AIDS was detected between cases and controls . Exposure to fluconazole for more than 90 days reduced the risk of cryptococcal meningitis by 82% (OR=0.18; 95% CI=0.04-0.85; p=0.03) . We did not find a difference in steroid use between cases and controls for either the length or amount of steroid exposure (p=0.41) . No difference in survival during follow-up in the clinic was observed by the log-rank test (p=0.74) . Among the cases, a cryptococcal antigen was positive in more than 97% of the CSF or blood samples . CSF and blood cultures were positive in 81 and 44% of the samples, respectively . We conclude that demographic factors did not affect the risk of cryptococcal meningitis in an inner city United States population . While fluconazole use has a protective effect, steroid use was not associated with an increased risk of cryptococcal meningitis in HIV-infected patients.

Mol Cell Biol, 1999 Jun, 19(6), 4101 - 12
Rapamycin antifungal action is mediated via conserved complexes with FKBP12 and TOR kinase homologs in Cryptococcus neoformans; Cruz MC et al.; Cryptococcus neoformans is a fungal pathogen that causes meningitis in patients immunocompromised by AIDS, chemotherapy, organ transplantation, or high-dose steroids . Current antifungal drug therapies are limited and suffer from toxic side effects and drug resistance . Here, we defined the targets and mechanisms of antifungal action of the immunosuppressant rapamycin in C . neoformans . In the yeast Saccharomyces cerevisiae and in T cells, rapamycin forms complexes with the FKBP12 prolyl isomerase that block cell cycle progression by inhibiting the TOR kinases . We identified the gene encoding a C . neoformans TOR1 homolog . Using a novel two-hybrid screen for rapamycin-dependent TOR-binding proteins, we identified the C . neoformans FKBP12 homolog, encoded by the FRR1 gene . Disruption of the FKBP12 gene conferred rapamycin and FK506 resistance but had no effect on growth, differentiation, or virulence of C . neoformans . Two spontaneous mutations that confer rapamycin resistance alter conserved residues on TOR1 or FKBP12 that are required for FKBP12-rapamycin-TOR1 interactions or FKBP12 stability . Two other spontaneous mutations result from insertion of novel DNA sequences into the FKBP12 gene . Our observations reveal that the antifungal activities of rapamycin and FK506 are mediated via FKBP12 and TOR homologs and that a high proportion of spontaneous mutants in C . neoformans result from insertion of novel DNA sequences, and they suggest that nonimmunosuppressive rapamycin analogs have potential as antifungal agents.

J Clin Microbiol, 1999 Jun, 37(6), 1732 - 8
Epidemiology of visceral mycoses: analysis of data in annual of the pathological autopsy cases in Japan; Yamazaki T et al.; The data on visceral mycoses that had been reported in the Annual of the Pathological Autopsy Cases in Japan from 1969 to 1994 by the Japanese Society of Pathology were analyzed epidemiologically . The frequency of visceral mycoses among the annual total number of pathological autopsy cases increased noticeably from 1.60% in 1969 to a peak of 4.66% in 1990 . Among them, the incidences of candidiasis and aspergillosis increased the most . After 1990, however, the frequency of visceral mycoses decreased gradually . Until 1989, the predominant causative agent was Candida, followed in order by Aspergillus and Cryptococcus . Although the rate of candidiasis decreased by degrees from 1990, the rate of aspergillosis increased up to and then surpassed that of candidiasis in 1991 . Leukemia was the major disease underlying the visceral mycoses, followed by solid cancers and other blood and hematopoietic system diseases . Severe mycotic infection has increased over the reported 25-year period, from 6.6% of the total visceral mycosis cases in 1969 to 71% in 1994 . The reasons for this decrease of candidiasis combined with an increase of aspergillosis or of severe mycotic infection might be that (i) nonsevere (not disseminated) infections were excluded from the case totals, since they have become controllable by antifungal drugs such as fluconazole, but (ii) the available antifungal drugs were not efficacious against severe infections such as pulmonary aspergillosis, and (iii) the number of patients living longer in an immunocompromised state had increased because of developments in chemotherapy and progress in medical care.

Planta Med, 1999 Apr, 65(3), 204 - 8
In vitro antifungal susceptibilities of Candida albicans and other fungal pathogens to polygodial, a sesquiterpene dialdehyde; Lee SH et al.; In vitro antifungal activities of polygodial were investigated against several fungal pathogens . Polygodial showed strong antifungal activity, comparable to amphotericin B, against yeast-like fungi Candida albicans, C . utilis, C . krusei, Cryptococcus neoformans, Saccharomyces cerevisiae and also filamentous fungi including Trichophyton mentagrophytes, T . ruburum, and Penicillium marneffei . Other strains such as Aspergillus fumigatus, A . flavus, P . chrysogenum, C . lipolytica, and C . tropicalis showed moderate to low susceptibility to polygodial . The anti-fungal activity of polygodial was generally not reduced by several susceptibility-testing conditions such as medium, incubation temperature, inoculum size, and medium pH . However, polygodial's antifungal activity was strongly increased at acidic conditions . Unlike amphotericin B, polygodial did not show any hemolytic activity and also its antifungal activity was not diminished in the presence of ergosterol . Based on killing kinetics against growing and nongrowing C . albicans, polygodial showed strong and rapid fungicidal activity.

Med Clin (Barc), 1999 Mar 27, 112(11), 401 - 5
{Disseminated cryptococcosis in patients with AIDS . Prognostic factors of poor outcome}; Alonso M et al.; BACKGROUND: The objectives of this study were to analyze the changes in the incidence of cryptococcal disease in the last 10 years (1987-1997) and to assess the factors of poor prognosis in HIV-1 infected patients . PATIENTS AND METHODS: Clinical records of HIV-1 infected patients diagnosed with cryptococcal infection from June 1987 to December 1997 at Hospital Clinic i Provincial in Barcelona, Spain, were examined . An univariate and multivariate analysis of the predictors of poor outcome was performed . RESULTS: Sixty clinical records were analyzed . The number of cases per 100 exposed patients per year decreased from 1991 to 1993 and, afterwards, decreased again from 1996 . Fifty patients had a resolution of clinical symptoms, 17 out of this 50 patients (34%) had a relapse . Factors associated with a higher risk of relapse were a positive blood culture and cryptococcal antigen title in cerebrospinal fluid above 1/1.024 . CONCLUSIONS: The decrease in the number of cases of cryptococcal infection coincides with the broad use of triazole antifungal drugs for oral candidiasis (1990-1991) and, afterwards, with the initiation of highly active antiretroviral therapy (1996) . The best predictors of relapse are a positive blood cryptococcal culture and a high titter of cryptococcal antigen.

Chemosphere, 1999 Jun, 38(13), 3031 - 9
Biodegradation of fluoranthene by soil fungi; Salicis F et al.; A selection of 39 strains of micromycetes known as good degraders of polychlorinated aromatic compounds, mostly isolated from soil and belonging to various taxonomic groups, have been investigated for fluoranthene degradation . Toxicity assays, first evaluated on solid medium MEA, have not shown any toxicity of fluoranthene (1-100 mg.L-1) towards fungi . Whereas, consumption assays on a solid synthetic medium showed a toxicity at 100 mg.L-1 . The degradation of fluoranthene (10 mg.L-1) was then investigated in a liquid synthetic medium for 4 days and evaluated by HPLC . Among the 39 strains tested, 18 degraded fluoranthene at 60% or more . Zygomycetes appeared to be the most efficient group (mean degradation: 90%) . Among 18 performant strains, 10 had not yet been reported in the literature: Sporormiella australis, Cryptococcus albidus, Cicinobolus cesatii, Pestalotia palmarum, beauveria alba, Aspergillus terreus . Cunninghamella blakesleeana, C . echinulata, Mortierella ramanniana and Rhizopus arrhizus . Fluoranthene adsorption on fungi was very low for the strains which degraded well fluoranthene (mean adsorption: 4%) . Whereas, some strains adsorbed it much more such as Colletotrichum dematium (47%) and Penicillium italicum (43%).

Biochem J, 1999 May 15, 340 ( Pt 1), 25 - 32
Inositol acylation of glycosylphosphatidylinositols in the pathogenic fungus Cryptococcus neoformans and the model yeast Saccharomyces cerevisiae; Franzot SP et al.; Cryptococcus neoformans, an opportunistic fungus responsible for life-threatening infection in immunocompromised patients, is able to synthesize glycosylphosphatidylinositol (GPI) structures . Radiolabelling experiments in vitro with the use of a cryptococcal cell-free system showed that the pathway begins as in other eukaryotes, with the addition of N-acetylglucosamine to phosphatidylinositol, followed by deacetylation of the sugar residue . The third step, acylation of the inositol ring, seemed to involve a fatty acid other than palmitate, in contrast with previous findings in Saccharomyces cerevisiae and mammalian GPI pathways . A systematic study of inositol acylation in C . neoformans and S . cerevisiae showed that both organisms used a variety of fatty acids in this step; these were transferred directly from acyl-CoA to inositol without modification . However, the specificity of fatty acid utilization was quite distinct in the two fungi, with the pathogen being substantially more restrictive . In mammalian cells fatty acids added exogenously as acyl-CoAs are not transferred directly to inositol . These results suggest significant differences in the GPI biosynthetic pathway between mammalian and C . neoformans cells that could represent targets for anti-cryptococcal therapy.

Radiology, 1999 May, 211(2), 507 - 12
Intracranial mass lesions: sequential thallium and gallium scintigraphy in patients with AIDS; Lee VW et al.; PURPOSE: To determine the efficacy of sequential thallium and gallium scintigraphy to differentiate intracranial neoplasms (lymphoma and glioma) from other nonmalignant intracranial mass lesions among patients with acquired immunodeficiency syndrome (AIDS) . MATERIALS AND METHODS: The authors reviewed the cases of 40 patients with human immunodeficiency virus (HIV) who underwent thallium and gallium scanning to evaluate intracranial mass lesions from October 1991 through November 1997 . There was a definitive final diagnosis of the nature of the mass lesions in 21 of these cases . In these 21 cases, the scintigraphic patterns were reviewed and were compared with the final diagnosis . RESULTS: On the basis of results at thallium and gallium scanning, the patients were divided into three groups . Group A included 13 patients (11 with brain tumors {lymphomas and gliomas} and two with progressive multifocal leukoencephalopathy {PML}) with thallium-positive, gallium-positive scans . Group B included five patients with intracranial infections (tuberculosis, Cryptococcus, bacteria) with thallium-negative, gallium-positive scans . Group C included three patients (one with PML and two with infarcts) with thallium-negative, gallium-negative scans . All patients with lymphomas were in group A . The sensitivity and specificity of the thallium-positive, gallium-positive pattern for intracranial malignancy were 100% and 80%, respectively . CONCLUSION: Sequential thallium and gallium scanning helped differentiate tumors from nonmalignant intracranial mass lesions and may help differentiate infections from PML or infarcts.

Int J Infect Dis, 1998-99 Winter, 3(2), 88 - 93
Etiology of central nervous system infections in the Philippines and the role of serum C-reactive protein in excluding acute bacterial meningitis; Sutinen J et al.; OBJECTIVES: The value of measurements of serum C-reactive protein (CRP) in differentiating central nervous system (CNS) infections of varying etiologies in the Philippines was investigated . METHODS: A wide array of bacteriologic and virologic methods as well as computed tomography, typical clinical presentation, and autopsy were used for etiologic diagnosis . RESULTS: Among 103 patients with CNS infection, etiology was identified in 60 (58%) cases . Bacteria were found in 19 (including 7 Streptococcus pneumoniae, 5 Haemophilus influenzae, 3 Neisseria meningitidis), tuberculosis in 4, viruses in 38 (including 20 coxsackievirus, 8 measles, 4 adenovirus, and 4 poliovirus infections), and brain abscess in 3 patients . C-reactive protein was elevated on admission in all 18 cases of bacterial meningitis tested, exceeding 50 mg/L in 17 (94%), and was not affected by prior antibacterial treatment . The mean CRP was significantly higher in the bacterial group than in the viral group (207 +/- 111 mg/L vs . 39 +/- 34 mg/L; P < 0.001) . In the viral group one third had CRP above 50 mg/L . In patients with tuberculous meningitis, brain abscess, or cryptococcal meningitis, CRP was moderately to highly elevated . CONCLUSIONS: In the presence of a normal CRP concentration (below 10 mg/mL) acute bacterial meningitis is excluded even in a developing country setting and antimicrobial therapy is not warranted.

Infect Immun, 1999 May, 67(5), 2357 - 65
Genetic and physiologic characterization of ferric/cupric reductase constitutive mutants of Cryptococcus neoformans; Nyhus KJ et al.; Cryptococcus neoformans is a pathogenic yeast that causes meningitis in immunocompromised patients . Because iron acquisition is critical for growth of a pathogen in a host, we studied the regulation of the ferric reductase and ferrous uptake system of this organism . We isolated 18 mutants, representing four independent loci, with dysregulated ferric reductase . The mutant strains had >10-fold higher than wild-type WT reductase activity in the presence of iron . Two of the strains also had >7-fold higher than WT iron uptake in the presence of iron but were not markedly iron sensitive . Both were sensitive to the oxidative stresses associated with superoxide and hydrogen peroxide . One strain exhibited only 23% of the WT level of iron uptake in the absence of iron and grew poorly without iron supplementation of the medium, phenotypes consistent with an iron transport deficiency; it was sensitive to superoxide but not to hydrogen peroxide . The fourth strain had high reductase activity but normal iron uptake; it was not very sensitive to oxidative stress . We also demonstrated that the ferric reductase was regulated by copper and could act as a cupric reductase . Sensitivity to oxidants may be related to iron acquisition by a variety of mechanisms and may model the interaction of the yeast with the immune system.

Infect Immun, 1999 May, 67(5), 2218 - 24
Antibody response to Cryptococcus neoformans proteins in rodents and humans; Chen LC et al.; The prevalence and specificity of serum antibodies to Cryptococcus neoformans proteins was studied in mice and rats with experimental infection, in individuals with or without a history of potential laboratory exposure to C . neoformans, human immunodeficiency virus (HIV)-positive individuals who developed cryptococcosis, in matched samples from HIV-positive individuals who did not develop cryptococcosis, and in HIV-negative individuals . Rodents had little or no serum antibody reactive with C . neoformans proteins prior to infection . The intensity and specificity of the rodent antibody response were dependent on the species, the mouse strain, and the viability of the inoculum . All humans had serum antibodies reactive with C . neoformans proteins regardless of the potential exposure, the HIV infection status, or the subsequent development of cryptococcosis . Our results indicate (i) a high prevalence of antibodies reactive with C . neoformans proteins in the sera of rodents after cryptococcal infection and in humans with or without HIV infection; (ii) qualitative and quantitative differences in the antibody profiles of HIV-positive individuals; and (iii) similarities and differences between humans, mice, and rats with respect to the specificity of the antibodies reactive with C . neoformans proteins . The results are consistent with the view that C . neoformans infections are common in human populations, and the results have implications for the development of vaccination strategies against cryptococcosis.

Oral Surg Oral Med Oral Pathol Oral Radiol Endod, 1999 Apr, 87(4), 437 - 41
Frequency of oropharyngeal candidiasis in HIV-infected patients on protease inhibitor therapy; Diz Dios P et al.; OBJECTIVE: The purpose of this study was to investigate the effect of HIV-1 protease inhibitors on the frequency of oropharyngeal candidiasis in HIV-infected patients . STUDY DESIGN: A clinical and analytic follow-up was carried out to determine the number of episodes of oropharyngeal candidiasis during HIV-1 protease inhibitor therapy and the relation of this incidence to the CD4 lymphocyte count and circulating neutrophils level . Seventy-five HIV-positive patients were selected, and HIV-1 protease inhibitor therapy was administered to each patient over a minimum of 6 months . These patients did not receive long-term preventive antifungal therapy for oral candidiasis, even as secondary prophylaxis against cryptococcosis . Results were compared with those obtained during the previous 6 months, during which patients had been treated only with reverse transcriptase inhibitors . RESULTS: At least one episode of oropharyngeal candidiasis was seen in 56% (42/75) of patients during reverse transcriptase inhibitor therapy and in only 9.3% (7/75) of patients after the initiation of protease inhibitor therapy . The number of relapses decreased significantly when the 2 follow-up periods were compared (P<.0001) . The CD4 and CD8 lymphocyte counts increased significantly with protease inhibitor therapy (P<.001 and P<.05, respectively) . During reverse transcriptase inhibitor treatment, the probability of the presentation of oropharyngeal candidiasis correlated with falling CD4 counts (P<.0001) . The HIV-1 protease inhibitor therapy was associated with a significant increase in the neutrophil count (P<.01) . The probability of the occurrence of some episode of candidiasis correlated inversely with the circulating neutrophil level (P<.05) . CONCLUSIONS: Protease inhibitor therapy decreases the frequency of HIV-related oropharyngeal candidiasis . The mechanism involved is unknown, but it can be speculated that a reduction of the viral load increases the number of intact T helper cells, which in turn enhances the number of circulating polymorphonuclear neutrophils and regulates their function by means of colony-stimulating factors.

Scand J Infect Dis, 1998, 30(6), 615 - 6
HIV combination therapy: immune restitution causing cryptococcal lymphadenitis dramatically improved by anti-inflammatory therapy; Blanche P et al.; Two patients with AIDS developed microscopically verified focal cryptococcal lymphadenitis while treated with highly active anti-retroviral therapy for 8 and 15 months . Both were treated with fluconazole as a secondary prophylaxis for prior cryptococcal meningitis . Cryptococcus neoformans did not grow . Amphotericin was ineffective . Anti-inflammatory drugs had a dramatic effect.

Genetics, 1999 May, 152(1), 167 - 78
Topoisomerase I is essential in Cryptococcus neoformans: role In pathobiology and as an antifungal target; Del Poeta M et al.; Topisomerase I is the target of several toxins and chemotherapy agents, and the enzyme is essential for viability in some organisms, including mice and drosophila . We have cloned the TOP1 gene encoding topoisomerase I from the opportunistic fungal pathogen Cryptococcus neoformans . The C . neoformans topoisomerase I contains a fungal insert also found in topoisomerase I from Candida albicans and Saccharomyces cerevisiae that is not present in the mammalian enzyme . We were unable to disrupt the topoisomerase I gene in this haploid organism by homologous recombination in over 8000 transformants analyzed . When a second functional copy of the TOP1 gene was introduced into the genome, the topoisomerase I gene could be readily disrupted by homologous recombination (at 7% efficiency) . Thus, topoisomerase I is essential in C . neoformans . This new molecular strategy with C . neoformans may also be useful in identifying essential genes in other pathogenic fungi . To address the physiological and pathobiological functions of the enzyme, the TOP1 gene was fused to the GAL7 gene promoter . The resulting GAL7::TOP1 fusion gene was modestly regulated by carbon source in a serotype A strain of C . neoformans . Modest overexpression of topoisomerase I conferred sensitivity to heat shock, gamma-rays, and camptothecin . In contrast, alterations in topoisomerase I levels had no effect on the toxicity of a novel class of antifungal agents, the dicationic aromatic compounds (DACs), indicating that topoisomerase I is not the target of DACs . In an animal model of cryptococcal meningitis, topoisomerase I regulation was not critically important to established infection, but may impact on the initial stress response to infection . In summary, our studies reveal that topoisomerase I is essential in the human pathogen C . neoformans and represents a novel target for antifungal agents.

Antimicrob Agents Chemother, 1999 May, 43(5), 1258 - 9
In vitro susceptibilities of clinical yeast isolates to the new antifungal eberconazole compared with their susceptibilities to clotrimazole and ketoconazole; Torres-Rodriguez JM et al.; The antifungal activity of eberconazole, a new imidazole derivative, against 124 clinical isolates of Candida comprising eight different species and to 34 isolates of Cryptococcus neoformans was compared to those of clotrimazole and ketoconazole . MICs of eberconazole, determined by the National Committee for Clinical Laboratory Standards standardized microbroth method, were equal to or lower than those of other azoles, especially for Candida krusei and Candida glabrata, which are usually resistant to triazoles.

Prostate, 1999 May, 39(2), 119 - 22
Primary cryptococcal prostatitis in an apparently uncompromised host; Caballes RL et al.; BACKGROUND: Systemic spread from a primary focus of cryptococcal infection commonly involves the central nervous system, manifested as meningitis or meningoencephalitis . Untreated meningitis and meningoencephalitis are invariably fatal, following a subacute or chronic course of cyclic remission and relapse, followed by progressive deterioration over weeks to months . Occasional patients with fulminating meningoencephalitis die within a few days . Incorrect diagnosis may be the most common cause of fatality in this disease . Thorough histopathologic examination and blood and body fluid studies should minimize misdiagnosis . METHODS: We studied the case of an initially misdiagnosed 64-year-old apparently immunologically competent man with primary cryptococcal prostatitis that subsequently disseminated to the central nervous system and the left eye . All immunologic findings including workup for AIDS viruses were normal . Laboratory studies confirmed cryptococcal infection . RESULTS: After initial misdiagnosis, our patient received improper treatment for 10 months . He developed meningitis and severe left endophthalmitis with optic nerve and retinal involvement . Toxic medications led to kidney insufficiency with about 66% loss of function . Following therapy, reevaluation of his immune system showed marked abnormality in cell-mediated immunity . CONCLUSIONS: Cryptococcosis is easily misdiagnosed in uncompromised hosts, both clinically and pathologically, because of misconception that the disease affects only immunocompromised individuals and that primary cryptococcal prostatitis is virtually unheard-of in "normal" males.

Br J Ophthalmol, 1999 May, 83(5), 535 - 9
Aetiological study of the presumed ocular histoplasmosis syndrome in the Netherlands; Ongkosuwito JV et al.; AIM: To investigate whether presumed ocular histoplasmosis syndrome in the Netherlands is caused by Histoplasma capsulatum and whether other risk factors might play a role in the pathogenesis of this syndrome . METHODS: 23 patients were clinically diagnosed as having presumed ocular histoplasmosis syndrome based on the following criteria: peripapillary atrophy, punched out lesions, a macular disciform lesion or scar in one eye without vitritis . As controls, 66 sex and age matched healthy volunteers were used . Serum samples from both patients and controls were tested for the presence of antibodies against H capsulatum, Toxoplasma gondii, Toxocara canis et cati, Ascaris sp, and for the presence of antigens of Cryptococcus neoformans . Serum samples were also tested for the presence of autoantibodies against retinal or choroidal proteins . To investigate other risk factors, patients and controls were asked to fill in a health and travel related questionnaire . Ten patients with ocular toxoplasmosis were used as a disease control group . RESULTS: None of the patients with presumed ocular histoplasmosis syndrome or controls had circulating antibodies directed against H capsulatum . No risk factors could be identified and no indications for autoimmunity and no evidence for the role of the other infectious agents could be demonstrated . CONCLUSIONS: In a Dutch group of patients fulfilling the criteria of a disease currently named presumed ocular histoplasmosis syndrome, no risk factors or relation with the fungus H capsulatum could be detected.

Bull Soc Pathol Exot, 1999 Feb, 92(1), 23 - 6
{Neurologic manifestations associated with HIV infections at the Bobo-Dioulasso Hospital Center (Burkina Faso)}; Millogo A et al.; Neurological manifestations of HIV infection are frequent and diverse . Diagnosis is often difficult due to geographical factors . 686 of the 3409 patients admitted to the Internal Medicine ward of Bobo-Dioulasso in 1995-1996 were HIV seropositive . This represents a prevalence of 20.1% . The sociodemographic and clinical characteristics of 101 patients with neurological problems during the study period are reported in this paper . This case series represents 14% of the HIV-positive admissions . The mean age was 35.7 years and 43% of the cases were aged 30 to 40 years . Sex-ratio was 1.6 male for 1 female . Focal deficits were observed in 36 of cases . Peripheral neuropathy (37%), meningitis or meningoencephalitis (20%), fitting (8%) and myelitis (8%) were the other clinical presentations . The etiology of the focal deficits was not ascertained because of the lack of tomodensitometry, specific serology and necropsy . Any neurological manifestation in a HIV seropositive patient should prompt investigations in order to diagnose those infections which can be treated, especially Toxoplasma gondii abscess and Cryptococcus neoformans meningitis.

Nihon Kokyuki Gakkai Zasshi, 1999 Feb, 37(2), 108 - 14
{Clinical investigation of 15 patients with pulmonary cryptococcosis--clinical comparison of HTLV-I carriers and non-carriers}; Matsuyama W et al.; We investigated 15 pulmonary cryptococcosis patients . The group had a mean age of 51.7 years, and 6 (40%) of the patients were women . Cryptococcosis was primary in 13 patients and secondary in 2 (diabetes mellitus and smoldering adult T-cell leukemia) . Eight patients were asymptomatic and 9 patients were detected by medical examinations . Dry cough was the most common symptoms . On chest radiographs, 5 patients showed solitary nodules, 4 patients showed infiltrative shadows, and 4 patients showed multiple nodules . The right lower lobe was the predominant location of solitary nodules, and the left upper and middle lung fields were the predominant locations of infiltrative shadows . Transbronchial lung biopsy was the method of diagnosis for 9 patients, and open lung biopsy for the others . Eleven patients were treated with fluconazole, and the mean treatment period was 7 months . Four patients underwent, resection procedures only, and experienced no recurrence . Five patients were positive for HTLV-I (one had smoldering ATL) and 5 were negative . Eighty percent of the HTLV-I positive patients had some symptoms and 80% of the HTLV-I negative patients were asymptomatic . HTLV-I positive patients showed various pulmonary shadows and 80% of the HTLV-I negative patients showed solitary nodules . The pulmonary lesions in HTLV-I positive patients were more extensive than those in HTLV-I negative patients (p < 0.05) . We postulate the possible existence of subtle immunological abnormalities, including abnormalities of cellular immunity, in HTLV-I carriers.

Kansenshogaku Zasshi, 1999 Feb, 73(2), 187 - 90
{A case of pulmonary cryptococcosis with lung cancer in a pulmonary lobe}; Yamamoto Y et al.; A 63-year old female was admitted because of an abnormal shadow on chest X-ray film . Chest CT showed a nodular shadow in the right S6 and a patchy shadow in the right S10 . Right lower lobectomy was performed under a diagnosis of lung cancer made by TBLB in the right S6 . Pathological examination of the resected lung revealed papillary adenocarcinoma in the right S6 and numerous cryptococci in the right S10 . No cryptococcal infection was found in the resected lymph nodes.

J Am Vet Med Assoc, 1999 Apr 15, 214(8), 1205 - 7, 1200
Bartonella spp infection as a possible cause of uveitis in a cat; Lappin MR et al.; A 6-year-old castrated mixed-breed cat was evaluated because of unilateral anterior uveitis . The cat was seronegative for antibodies to Toxoplasma gondii, coronaviruses, and feline immunodeficiency virus, and antigens for FeLV p27 and Cryptococcus neoformans . Antibodies to Bartonella spp were detected in serum and aqueous humor . The antibody coefficient (C value) for IgG antibodies to Bartonella spp in the aqueous humor was 4.42; values > 1 suggest ocular production of antibodies and supports a diagnosis of ocular infection . Topical administration of prednisolone and oral administration of prednisone failed to induce a response; however, the uveitis resolved rapidly after the cat was given doxycycline orally . Clinical or laboratory evidence of immunodeficiency in this cat was not detected . Detection of a serum IgG antibody titer to Bartonella spp and ocular production of IgG antibodies to Bartonella spp, exclusion of other causes of uveitis, and response to doxycycline suggests that the cat may have had bartonellosis resulting in uveal tract inflammation.

Eur J Dermatol, 1999 Apr-May, 9(3), 224 - 6
Primary cutaneous cryptococcosis in an HIV-negative patient; Bellosta M et al.; We report a case of primary cutaneous cryptococcosis in an HIV-negative patient, who presented with painless, ulcerated lesions involving the right forearm (fingers and elbow), which developed over 45 days . On the basis of the clinical appearance, serological and cultural examinations were performed to confirm the diagnosis; the histological evaluation of a skin biopsy showed an acute inflammatory infiltrate containing several PAS + Cryptococci . Subsequently, the patient was treated with fluconazole (400 mg/day for 10 days, then a maintenance therapy of 200 mg/day); after one month, the cutaneous lesions were remarkably improved, but, although a series of further laboratory and clinical examinations was scheduled, the patient repeatedly refused any other re-evaluation, and he was lost from follow-up.

Head Neck, 1999 May, 21(3), 239 - 46
Supraclavicular masses: results of a series of 309 cases biopsied by fine needle aspiration; Ellison E et al.; BACKGROUND: The purpose of this study was to determine the current distribution of diseases resulting in supraclavicular swelling or lymphadenopathy as diagnosed by fine needle aspiration (FNA) biopsy . METHODS: Supraclavicular aspirates in this retrospective 5-year study from a large public hospital were classified as neoplastic, infectious, inflammatory, reactive, and nondiagnostic . RESULTS: Malignancy was present in 55% of the 309 aspirates (47% metastatic, 8% lymphoma) . Age was most predictive of malignancy (32% for age < or =41 years, 68% for age >40 years) . Lymphoma occurred equally in both groups, but the lymphoma:metastasis ratio was much higher in younger patients (1:1.6 for age < or =41 years versus 1:11 for age >40 years) . Ethnic origin was related to tumor type, metastatic uterine cervical carcinoma being most frequent in Hispanics, and lymphoma in Caucasians . Primary oropharyngeal sites were unusual; most malignancies originated in the lung, breast, or cervix . Left or right side did not discriminate for either the presence or type of tumor . The 43 aspirates from human immunodeficiency virus (HIV) positive (+) patients were predominantly inflammatory, infectious, or reactive; lymphoma was infrequent (7%) . Overall, 39 patients had mycobacterial infection; aspirate smears or culture were positive in 19 . Three other FNAs grew Staphylococcus aureus and one stained for Cryptococcus . Aspirates were not informative in 24 cases (8%) . The sensitivity of FNA for malignancy was 97%, specificity was 98%, and positive predictive value 98% . CONCLUSION: Aspiration biopsy is an excellent diagnostic tool for supraclavicular masses . Patients over 40 years old are likely to have metastatic malignancy, from breast, lung, or infradiaphragmatic sites . Aspirates with inflammation and those from HIV+ patients should undergo extensive culture.

J Neuroimaging, 1999 Apr, 9(2), 118 - 21
Pontine cryptococcoma in a nonimmunocompromised individual: MRI characteristics; Kesler R et al.; The case of a pontine cryptococcoma in a nonimmunocompromised, previously healthy 16-year-old boy is presented . The patient had slowly progressive brainstem signs with right cranial nerves V, VII, and VIII palsies, and contralateral corticospinal and spinothalamic deficits . Magnetic resonance images (MRI) revealed, within the right pons, a 1-cm diameter round mass lesion, hypointense on T1-weighted images, hyperintense on T2-weighted images, and with rim enhancement after infusion of gadopentetate dimeglumine . This is the only report of the MRI findings in an isolated pontine cryptococcoma in an immunocompetent patient . Early recognition of this specific MRI pattern is essential, because complete recovery can be achieved with prompt antifungal treatment.

AIDS, 1999 Jan 14, 13(1), 49 - 55
Granulocyte-macrophage colony-stimulating factor and fluconazole enhance anti-cryptococcal activity of monocytes from AIDS patients; Tascini C et al.; OBJECTIVE: To investigate the effect of human recombinant granulocyte-macrophage colony-stimulating factor (hrGM-CSF) and fluconazole on anti-cryptococcal activity of monocytes from AIDS patients and normal subjects . DESIGN: The effect of GM-CSF and fluconazole on fungistatic and fungicidal activity of monocytes was studied in an in vitro system . METHODS: Monocytes were treated in vitro with hrGM-CSF and fluconazole or either agent alone for 24 or 48 h, and fungistatic and fungicidal activity was evaluated in a colony-forming unit inhibition assay . CD11b/CD18 expression in monocytes was measured by flow cytometry analysis . Superoxide anion generation by peripheral blood monocytes was measured in the presence of pre-opsonized zymosan . RESULTS: Defective antifungal capacity of monocytes from AIDS patients was observed . GM-CSF treatment of monocytes from AIDS patients increased fungistatic activity, and the combination of hrGM-CSF and fluconazole resulted in fungicidal activity . The mechanisms involved in the GM-CSF-mediated effect appeared to be mediated by (i) enhancement of phagocytic activity, (ii) increase of superoxide anion generation, and (iii) upregulation of CD11b/CD18 expression on the monocyte surface . CONCLUSIONS: Our data highlight the effect of GM-CSF on anti-cryptococcal activity of human monocytes and show a synergistic effect of GM-CSF with fluconazole, suggesting a new therapeutic strategy in the treatment of cryptococcosis.

Med Mycol, 1998 Dec, 36(6), 419 - 24
Pathogenicity of basidiospores of Filobasidiella neoformans var . neoformans; Sukroongreung S et al.; Basidiospores of Filobasidiella neoformans var . neoformans (progeny of Cryptococcus neoformans MT 100.1 x VR 45980) were able to induce cryptococcosis in Swiss albino mice if inoculated by intraperitoneal injection, nasal instillation or nasal spraying . The latter method, with the aid of a jet nebulizer, was first adopted to imitate the natural entrance of infectious particles . Using this method the small number of basidiospores (7 x 10(3)) could induce cryptococcosis in mice, while the higher number of the parental laboratory-grown yeast cells (1.5 x 10(6)) did not produce infections . By nasal instillation Cyclophosphamide (Cy)-treated mice were more susceptible to the basidiospores, showing 80% cryptococcosis (eight of 10) . Seven of the eight infected mice had disseminated cryptococcosis . Immunocompetent mice were more resistant to basidiospore infection than Cy-treated mice, as 40% of that group developed only pulmonary cryptococcosis; none had disseminated infection . Thus, we propose that basidiospores are one form of the infectious propagules of F . neoformans var . neoformans which can cause cryptococcosis, particularly in immunocompromised people.

Med Mycol, 1998 Dec, 36(6), 391 - 4
Isolation of Cryptococcus neoformans var . neoformans from pigeon droppings collected throughout Turkey; Yildiran ST et al.; The six hundred and thirty-four samples of pigeon droppings were collected throughout Turkey, from 54 of 80 provinces . Cryptococcus neoformans was isolated from 29 (4.6%, overall) of 634 samples and 29 isolates were from 18 provinces . Interestingly, 16 (88.9%) of these provinces occur on the three different coastlines of Turkey, therefore the ecological role of a humid climate was speculated . Almost all isolates {28/29} were recovered from samples collected from roofs (n=14) and dovecotes (n=14) . All isolates were found to be C . neoformans var . neoformans.

J Hosp Infect, 1999 Mar, 41(3), 181 - 94
Invasive yeast infections other than Candida spp . in acute leukaemia; Krcmery V et al.; During the last two decades, the treatment of leukaemia has changed significantly; increasing intensity of chemotherapy and bone marrow transplantation have lead to profound immunosuppression, prolonged stays in hospital, vascular catheterization, administration of broad spectrum antimicrobial agents and extensive use of prophylactic antifungal drugs . All but the last of these risk factors have increased the incidence of fungal infections in leukaemic patients and have significantly changed the spectrum of pathogens in favour of non-Candida species . In major haematological centres in Europe and the US, the proportion of non-Candida yeast isolated from patients increased from 1 to 5% in 1980 to 10 to 25% after 1990 . However, there are not enough data to assess whether mortality due to these species is higher than that due to Candida spp . (30-40%) or filamentous fungi (50-70%) . In this article, specific risk factors and therapeutic outcome of yeast infections other than Candida spp . in leukaemia such as Malassezia furfur, Trichosporon spp., Blastoschizomyces capitatus, Rhodotorula rubra, Saccharomyces cerevisiae, Clavispora lusitaniae, Cryptococcus laurentii and Hansenula anomala are reviewed . An analysis of risk factors from the National Cancer Institute, Bratislava has shown that non-Candida yeast infections (N = 15) in cancer patients are associated with leukaemia and neutropenia (P = < 0.002, 0.005), more often fatal than those caused by C . albicans (N = 51) (P < 0.006) but not non-albicans Candida (N = 34) and are associated with quinolone (P < 0.0001) and itraconazole prophylaxis (P < or = 0.05, 0.015) compared to both C . albicans or non-albicans Candida respectively.

J Med Primatol, 1998 Dec, 27(6), 298 - 302
Intestinal cryptococcosis in a common marmoset (Callithrix jacchus); Juan-Salles C et al.; A five-year-old female common marmoset (Callithrix jacchus) died after a one-month clinical course of nonspecific signs . Pathologic findings were acute diffuse fibrinonecrotizing enteritis and granulomatous endolymphangitis of intestinal and mesenteric lymphatic vessels . Both lesions were associated with a marked proliferation of Mayer's mucicarmine-positive, 4 to 15 microm yeasts that were surrounded by a wide clear halo . The infection was probably acquired by oral route . Other findings included moderate multifocal granulomatous and necrotizing hepatitis and mesangial nephropathy . Although the immunological status of this marmoset was unknown, cryptococcosis might induce primary lethal intestinal infections in callitrichids.

Cell Immunol, 1999 Apr 10, 193(1), 9 - 16
Role of TNF-alpha in the induction of fungicidal activity of mouse peritoneal exudate cells against Cryptococcus neoformans by IL-12 and IL-18; Kawakami K et al.; We have recently demonstrated that two IFN-gamma-inducing cytokines, interleukin (IL)-12 and IL-18, synergistically induced the fungicidal activity of mouse peritoneal exudate cells (PEC) against Cryptococcus neoformans through NK cell production of interferon (IFN)-gamma and nitric oxide (NO) synthesis . In the present study, we further dissected these effects by examining the involvement of tumor necrosis factor (TNF)-alpha in the induction of IL-12/IL-18-stimulated PEC fungicidal activity . The addition of neutralizing anti-TNF-alpha mAb significantly suppressed IL-12/IL-18-stimulated PEC anticryptococcal activity . This effect was ascribed to the inhibition of macrophage NO synthesis, but not of IFN-gamma production by NK cells, because the same treatment inhibited the former response, but not the latter one . On the other hand, combined treatment with IL-12 and IL-18 synergistically induced the production of TNF-alpha by PEC and this effect was almost completely abrogated by neutralizing anti-IFN-gamma mAb . The cell type producing TNF-alpha among PEC was mostly macrophage . TNF-alpha significantly promoted macrophage NO production and anticryptococcal activity induced by IFN-gamma, and furthermore anti-TNF-alpha mAb partially inhibited these responses . Considered together, our results indicated that TNF-alpha contributed to the potentiation of IL-12/IL-18-induced PEC fungicidal activity against C . neoformans through enhancement of IFN-gamma-induced production of NO by macrophages, but not through increased production of IFN-gamma by NK cells .

J Immunol, 1999 Apr 15, 162(8), 4824 - 33
Role of the C-C chemokine, TCA3, in the protective anticryptococcal cell-mediated immune response; Doyle HA et al.; Activated T lymphocytes play a crucial role in orchestrating cellular infiltration during a cell-mediated immune (CMI) reaction . TCA3, a C-C chemokine, is produced by Ag-activated T cells and is chemotactic for neutrophils and macrophages, two cell types in a murine CMI reaction . Using a gelatin sponge model for delayed-type hypersensitivity (DTH), we show that TCA3 is a component of the expression phase of an anticryptococcal CMI response in mice . TCA3 mRNA levels are augmented in anticryptococcal DTH reactions at the same time peak influxes of neutrophils and lymphocytes are observed . Neutralization of TCA3 in immunized mice results in reduced numbers of neutrophils and lymphocytes at DTH reaction sites . However, when rTCA3 is injected into sponges in naive mice, only neutrophils are attracted into the sponges, indicating TCA3 is chemotactic for neutrophils, but not lymphocytes . We show that TCA3 is indirectly attracting lymphocytes into DTH-reactive sponges by affecting at least one other chemokine that is chemotactic for lymphocytes . Of the two lymphocyte-attracting chemokines assessed, monocyte-chemotactic protein-1 and macrophage-inflammatory protein-1alpha (MIP-1alpha), only MIP-1alpha was reduced when TCA3 was neutralized, indicating that TCA3 affects the levels of MIP-1alpha, which attracts lymphocytes into the sponges . TCA3 also plays a role in protection against Cryptococcus neoformans in the lungs and brains of infected mice, as evidenced by the fact that neutralization of TCA3 results in increased C . neoformans CFU in those two organs.

Med Mycol, 1999 Feb, 37(1), 61 - 7
Molecular cloning of a second phospholipase B gene, caPLB2 from Candida albicans; Sugiyama Y et al.; Accumulating evidence suggests that phospholipase B, secreted by pathogenic fungi such as Candida albicans, Cryptococcus neoformans and Aspergillus fumigatus, functions as one of the virulence factors . In the present study, we have attempted to clone phospholipase B gene from C . albicans . By RT-PCR analysis with degenerate primers based on conserved regions of phospholipase B from Saccharomyces cerevisiae, Penicillium notatum and Torulaspora delbrueckii two similar but different cDNA fragments were obtained . One corresponded to the partial sequence of caPLB1, recently cloned phospholipase B gene from C . albicans by a different approach (Leidich et al.: J Biol Chem 1998; 273: 26078-86) . The other fragments contained sequences similar to the corresponding sequences of phospholipase B from other fungi . The presence of two related genes was confirmed by Southern and Northern blot analyses . The full length of the second C . albicans phospholipase B gene (caPLB2) encoded a putative protein with 608 amino acids and contained a potential signal peptide sequence and a putative catalytic region, which are found in phospholipase B from other fungi . Consistent with the findings of caPLB1, caPLB2 also lacks a cluster of hydrophobic amino acids at the COOH-terminal, which may function as a signal of glycosylphosphatidylinositol anchor.

Med Mycol, 1999 Feb, 37(1), 43 - 51
Serum antibody response to Cryptococcus neoformans in cats, dogs and koalas with and without active infection; Malik R et al.; Anti-cryptococcal antibodies were measured in normal cats, dogs, horses and koalas, and cats, dogs and koalas with cryptococcosis using an enzyme immunoassay . Antibody levels were expressed as absorbance readings . Over 80% of cats and dogs with cryptococcal infection had elevated antibody levels at the time of diagnosis, during or after successful therapy . Antibody levels in these patients either remained elevated or declined slowly after treatment . For cats, anti-cryptococcal antibody levels were higher in C . neoformans var . gattii than var . neoformans infections, and lower in mild than in moderate or severe infections . The persistence of increased anti-cryptococcal antibody levels in over half of the feline and canine cases following active infection suggested the use of antibody determinations as a seroepidemiologic marker of previous infection . Consequently, antibody measurements from 'normal' animals indicated a prevalence of previous cryptococcal infection of 10% in cats and dogs, compared with 3% in horses and 5% in koalas . Preliminary studies of young animals suggested that anti-cryptococcal antibody levels were substantially lower in the young cats but not the young dogs surveyed, compared with their mature counterparts . The cut-offs used in the present work may thus be erroneously high, with a corresponding underestimation of the prevalence of inapparent cryptococcosis.

Clin Infect Dis, 1999 Mar, 28(3), 629 - 33
Treatment of hydrocephalus secondary to cryptococcal meningitis by use of shunting; Park MK et al.; Hydrocephalus can be associated with increased morbidity and mortality in cryptococcal meningitis if left untreated . Both ventriculoperitoneal and ventriculoatrial shunting have been used in persons with cryptococcosis complicated by hydrocephalus, but the indications for and complications, success, and timing of these interventions are not well known . To this end, we reviewed the clinical courses of 10 non-human immunodeficiency virus-infected patients with hydrocephalus secondary to cryptococcal meningitis who underwent shunting procedures . Nine of 10 patients who underwent shunting had noticeable improvement in dementia and gait . Two patients required late revision of their shunts . Shunt placement in eight patients with acute infection did not disseminate cryptococcal infection into the peritoneum or bloodstream, nor did shunting provide a nidus from which Cryptococcus organisms proved difficult to eradicate . Shunting procedures are a safe and effective therapy for hydrocephalus in patients with cryptococcal meningitis and need not be delayed until patients are mycologically cured.

Clin Exp Immunol, 1999 Mar, 115(3), 485 - 90
Heat shock protein 70 (hsp70) as a major target of the antibody response in patients with pulmonary cryptococcosis; Kakeya H et al.; Cryptococcus neoformans causes infection in individuals with defective T cell function, such as AIDS, as well as without underlying disease . It has been suggested that humoral as well as cellular immunity might play an important role in the immune response to C . neoformans infection . We have recently shown, using immunoblotting, that the 70-kD hsp family of C . neoformans was the major target molecule of the humoral response in murine pulmonary cryptococcosis . In this study we also used immunoblotting to define the antibody responses in the sera of 24 patients with pulmonary cryptococcosis: 21 proven and three suspected diagnoses . Anti-C . neoformans hsp70 antibody was detected in 16 of 24 (66.7%) patients with pulmonary cryptococcosis . Fourteen of 17 (82.3%) patients with high antigen titres (> or = 1:8) and two of seven (28.6%) patients with low titres (< or = 1:4) had detectable levels of anti-hsp70 antibody . Sera from patients positive for anti-hsp70 antibody showed high titres in the Eiken latex agglutination test for the detection of serum cryptococcal antigen . Our results indicate that the 70-kD hsp family from C . neoformans appears to be a major target molecule of the humoral response, not only in murine pulmonary cryptococcosis, but also in human patients with pulmonary cryptococcosis.

J Clin Gastroenterol, 1999 Apr, 28(3), 273 - 5
Hepatobiliary dysfunction as the initial manifestation of disseminated cryptococcosis; Lin JI et al.; A case of hepatobiliary dysfunction as the initial manifestation of disseminated cryptococcosis is described . The patient was admitted with symptoms of hepatitis with cholestatic jaundice . Antibody tests for hepatitis B and C and human immunodeficiency virus were negative . The patient continued to deteriorate clinically . Eventually, the patient succumbed to hepatic failure . Autopsy disclosed systemic cryptococcosis that caused extensive necrosis of the liver . In review of the literature, only nine cases of cryptococcal infection presenting as hepatitis, cholangitis, and cholecystitis as initial manifestation were reported . Four of these patients had been subjected to exploratory laparotomy for clinical suspicion of acute abdomen . One patient developed cirrhosis as a result of cryptococcal hepatitis . Two patients succumbed to hepatic failure . Cryptococcosis is known to occur commonly in immunocompromised patients, yet only two reported cases presenting as hepatitis were associated with immunocompromised status.

J Neurovirol, 1999 Feb, 5(1), 76 - 81
Dissemination of C . neoformans to the central nervous system: role of chemokines, Th1 immunity and leukocyte recruitment; Huffnagle GB et al.; Cryptococcus neoformans is a fungus that possesses two properties unique for yeast: (1) production of a polysaccharide capsule and (2) neurotropism . The natural route of infection by C . neoformans is the respiratory tract; thus, factors that regulate the development and recruitment of memory Th1 cells and monocytes into the brain are critical for an effective response against disseminated C . neoformans infection . Production of TNFalpha prior to day 7 is required to prevent colonization of the central nervous system (CNS) . Th1 type immunity is required to clear established foci . In contrast, Th2 type immunity is ineffective at eliminating the infection in the brain and results in decreased survival . C . neoformans infection of MIP-1alpha and CCR5 knockout mice has highlighted the complex role that some chemokines may play in different organs . MIP-1alpha knockout mice have decreased leukocyte recruitment and cryptococcal clearance from the brain compared to wild-type mice . Thus, the host defence mechanisms that clear C . neoformans from the CNS appear to be similar to those in the lungs: via a Th1 cell-mediated inflammatory response that requires chemokines for the recruitment of effector cells.

Int J Gynaecol Obstet, 1999 Feb, 64(2), 177 - 80
Multiple hepatocellular adenomas in a patient with a history of oral contraception; Caballes RL et al.; We report a previously undescribed association between multiple hepatocellular adenomas and the use of oral contraceptives in a 29-year-old woman . She underwent trisegmentectomy, with 90% of the liver resected . Based on the rather large size of some of the adenomas at the time of diagnosis, most likely, the lesions were already present years before they were discovered . The liver was fully regenerated 6 months after surgery . Four years later, multiple adenomas recurred . During work-up for a liver transplant, she was found to have cryptococcal meningitis and pneumohydrocephalus which caused her demise.

Syst Appl Microbiol, 1999 Feb, 22(1), 97 - 105
Polyphasic taxonomy of a novel yeast isolated from antarctic environment; description of Cryptococcus victoriae sp . nov; Montes MJ et al.; In 1992 some samples of mosses, lichens and soils were collected from Botany Bay, Southern Victoria Land (77 degrees 01' S 162 degrees 32' E) and, as a result of a routine screening programme some yeasts were isolated . One of them, designated as strain G5, showed marked differences when compared to other antarctic yeasts . According to morphological and physiological characteristics, we were able to identify the strain G5 as a yeast belonging to the genus Cryptococcus . Some characteristics of this genus are the growth response to myo-inositol, celobiose, raffinose and D-glucuronate, no-fermentation, the absence of mycelium and pseudomycelium, asexual reproduction, Diazolium blue B test (DBB) and urea hydrolisis positive and the growth without vitamines . This strain (G5) formed cream colonies of slimy appearance with cells of 3 x 2 microns in size, that grew between 4 degrees C and 20 degrees C . The G + C content of strain G5 was 50.3 mol% . The molecular characterization by whole-cell proteins and RFLP analysis of the 5.8S rRNA gene and the two ribosomal internal transcribed spacers (5.85-ITS region), revealed that this strain was different from other antarctic species of this genus . The phylogenetic tree deduced from the 5.8S rRNA gene sequence showed the strain G5 as a member of the genus Cryptococcus, clearly separated from other basidiomycetous yeasts . On the basis of the physiological, genotypical and phylogenetical data, the new isolate G5 was described as Cryptococcus victoriae, sp . nov., with the type strain G5 (= CECT 11114).

Curr Opin Neurol Neurosurg, 1991 Apr, 4(2), 196 - 201
Neuroimaging in infections and demyelinating disease; Paty DW; Computerized tomography and magnetic resonance imaging continue to illuminate the changes that occur in the central nervous system in infections and demyelinating disease . Imaging in the acquired immune deficiency syndrome helps to better understand neurological complications . Magnetic resonance imaging also helps to be specific about the diagnosis of cryptococcal meningitis and toxoplasmosis . Major contributions have been made to the understanding of the diagnosis and the living pathology of multiple sclerosis . Experimental studies have identified the mechanism of blood-brain barrier disruption in inflammatory disease.

Hosp Formul, 1990 Mar, 25 Suppl B, 9 - 14
P&T Committee review of fluconazole: an effective alternative to antifungal therapy; Neu HC et al.; Fluconazole is a new antifungal agent available in both oral and parenteral formulations . According to the experts in this roundtable discussion, fluconazole represents a major clinical advance in the treatment of candidiasis and cryptococcosis in cancer patients, patients with AIDS, organ transplant recipients, and other patients at risk for opportunistic mycoses . The pharmacokinetic profile for fluconazole permits infrequent dosing and also makes it ideal for tissue site infections . Fluconazole's low toxicity gives it an advantage over currently available antifungal therapy and will permit prompt presumptive treatment of selected infections.

Ann Acad Med Singapore, 1998 Nov, 27(6), 873 - 6
Cryptococcal prostatic abscess in an immunocompromised patient: a case report and review of the literature; Yip SK et al.; A case of cryptococcal prostatic abscess in a 65-year-old Chinese man with immunosuppression from treatment of myasthenia gravis is presented . The patient was diagnosed to have cryptococcaemia when he presented with fever and urinary symptoms . Further investigations confirmed cryptococcal meningitis and imaging studies showed a hypodense lesion in the prostate . This proved to be an abscess and it was deroofed transurethrally . Histology of the prostatic tissue revealed the presence of Cryptococcus . The prostate can be a site of persistent cryptococcal infection and may take the form of an abscess . It should be drained transurethrally to prevent relapse.

Neurologia, 1999 Feb, 14(2), 62 - 6
{Meningitis in subjects with human immunodeficiency virus infection}; Silva-Rosas C et al.; BACKGROUND: Neurological complications are frequently observed in HIV-1 patients . Lumbar puncture (LP) and LSF analysis are two key diagnostic procedures . AIM: To describe the etiology meningeal syndromes in a hospital series of HIV patients . RESULTS: In this study, we present the different meningeal complications from 198 HIV-1 patients referred, for the last five years, to the HIV Center of the Hospital of University of Chile . The diagnosis of HIV-1 was done clinically plus a positive ELISA test, and confirmed by Western blot and/or PCR . In all cases with a possible neurological complication (52/198), a LP was performed . Cytochemical and microbiological studies, were done in each CSF sample . Serum CD4/CD8 lymphocytes number were determined by flow cytometry, and brain CT scan and/or MRI were obtained . From the 52 patients in whom a LP was done, 24 showed an abnormal CSF, compatible with the diagnosis of meningitis . The most frequent etiology (11/24) was infection by Cryptococcus neoformans, followed by Treponema pallidum (7/24) . There were 3 cases of HIV-1 meningitis, and 3 other cases with lymphoma, varicella zoster and cytomegalovirus meningitis . The frequency of cryptococcal infection was similar to that reported in the literature, but two interesting observations were the high frequency of neurosyphilis and the absence of TBC meningitis . In our country, the VDRL/FTA-ABS serum tests are mandatory in HIV patients . If these tests were positive a LP was performed, and this could partially explain the high number of cases coinfected with neurosyphilis . There is an important prevalence of lung TBC in our country, and as a consequence there is a policy of immunization to all newborn . It is possible that the high prevalence of TBC "promotes" a more actively search for TBC infection, with an early diagnosis and treatment of pulmonary TBC, and so a more frequent prophylaxis therapy in HIV patients, without the development of TBC meningitis.

Am J Ther, 1998 Jan, 5(1), 45 - 9
Management of opportunistic infections in acquired immunodeficiency syndrome . I . Treatment; Schlager SI; A case report of a patient infected with human immunodeficiency virus (HIV) is described . The patient presents with a multitude of medical complaints that are of acute or subacute onset . The medical examination of these complaints is described and includes algorithms for the diagnosis and treatment of the most common HIV-related opportunistic infections, including Pneumocystis carinii pneumonia, toxoplasmosis, Mycobacterium avium complex, cytomegalovirus infection, and cryptococcal meningitis.

Klin Med (Mosk), 1999, 77(1), 39 - 42
{Clinical course of cryptococcosis in HIV infection}; Rakhmanova AG et al.; Cryptococcosis was detected in 17% of examines with AIDS . The disease was caused by fungus Cryptococcus neoformans . Most frequently cryptococcosis affected CNS (meningitis and encephalitis) . The symptoms were scare and nonspecific, e.g . positive meningitis indicators occurred only in 9% of the patients . Cryptococcosis tends to hematogenic spread with severe dissemination . The diagnosis was made primarily basing on the results of mycological investigations of the biosubstrates, i.e . discovery of Cryptococcus neoformans.

Zh Mikrobiol Epidemiol Immunobiol, 1999 Jan-Feb, (1), 80 - 2
{The structure of the secondary diseases in HIV-infected patients in Russia}; Ermak TN et al.; In 95 patients with HIV infection (76 males and 19 females), observed in 1993-1997, the structure of secondary diseases was studied . During this period 58 patients (61.1%) died . In the structure of secondary diseases represented by 14 nosoforms prevailed cytomegalovirus and candidal infections, tuberculosis, Kaposi's sarcoma, rarely--pneumocystis pneumonia, cryptococcal meningitis and toxoplasmosis.

Eur J Pediatr, 1999 Mar, 158(3), 187 - 99
Current approaches to diagnosis and treatment of fungal infections in children infected with human immuno deficiency virus; Muller FM et al.; The prolonged survival of profoundly immunocompromised patients has revealed mucosal and invasive fungal infections to be major causes of morbidity and mortality in advanced HIV disease in children of all age groups . Antifungal resistance has become a clinically relevant problem . Paediatricians caring for HIV-infected children need to be aware of these increasingly frequent and often life-threatening infectious complications . This article reviews what is currently known about epidemiology, clinical presentation, diagnosis and treatment of mucosal and invasive fungal infections in children and adolescents with HIV disease . Candida spp . have become a leading bloodstream isolate in hospitalised patients; mucosal candidiasis is the most prevalent opportunistic infection in HIV-infected patients, and in both invasive and superficial infections, non Candida albicans spp . are on the increase . Invasive pulmonary aspergillosis has surfaced as an HIV-associated complication and previously uncommon fungi are more frequently encountered . HIV-infected individuals are particularly susceptible to Peumocystis carinii, Cryptococcus neoformans and infections by endemic fungi, such as Histoplasma capsulatum, Coccidioides immitis, Penicillium marneffei, and others . Newer immunological and molecular-based methods provide early and rapid diagnosis and monitoring . Potent and broad-spectrum third generation triazoles and novel fungicidal lipopeptides of the echinocandin class of antifungal antibiotics have entered clinical trials . Immunmodulation by recombinant cytokines and antifungal vaccines are very actively pursued inroads to adjunctive and preventive immunotherapy . CONCLUSION: Mucosal and invasive fungal infections will remain important complications in HIV-infected children of all age groups . Interventional studies and well documented case series are needed to improve the molecular diagnosis, treatment and prevention of invasive fungal infections in the paediatric HIV-infected population.

Eur J Clin Invest, 1999 Jan, 29(1), 83 - 92
Pulmonary surfactant protein A binds to Cryptococcus neoformans without promoting phagocytosis; Walenkamp AM et al.; BACKGROUND: Evidence is accumulating that the alveolar collecting surfactant protein A (SP-A) plays an important role in the first line of defence against infiltrating pathogenic micro-organisms and viruses . The ability of SP-A to facilitate the binding and uptake of acapsular Cryptococcus neoformans by monocyte-derived macrophages, human alveolar macrophages, monocytes and polymorphonuclear leucocytes was investigated . MATERIALS AND METHODS: Binding, competition and phagocytosis experiments were performed using a flow cytometry technique . RESULTS: SP-A bound to both the acapsular and the encapsulated form of C . neoformans in a concentration-dependent manner . SP-A showed a threefold better binding to the acapsular yeast: this binding was partly calcium dependent and could be inhibited by mannose (ID50 = 3 mmol L-1) and glucose (ID50 = 2.1 mmol L-1) but not by galactose (ID50 = 391 mmol L-1) . SP-A did not function as an opsonin in phagocytosis of acapsular C . neoformans for any of the phagocytes studied . CONCLUSION: Our results indicate that SP-A binds in a concentration-dependent manner to both encapsulated and acapsular C . neoformans . Despite SP-A binding to the acapsular C . neoformans, phagocytosis by various phagocytes was not enhanced.

Neuroradiology, 1999 Feb, 41(2), 129 - 33
Cryptococcus meningoencephalitis in AIDS: parenchymal and meningeal forms; Berkefeld J et al.; CT and MRI in one case of Cryptococcus neoformans infection showed contrast-enhancing parenchymal lesions resembling granulomata or abscesses . After an initial phase without contrast enhancement, the full extent of the lesions was visible within 2 weeks of presentation . The enhancing masses were assumed to represent intracerebral cryptococcomas . Despite evidence of massive meningeal infection on cerebrospinal fluid (CSF) examination, no radiological signs of meningitis, invasion of the Virchow-Robin spaces or ventriculitis could be demonstrated . With antimycotic treatment the contrast enhancement disappeared and cystic, partly calcified lesions remained . Recurrence of meningeal infection without radiological correlates was apparent in this stage . In a second case of proven cryptococcus meningitis, dilation of Virchow-Robin spaces or cysts in the adjacent parenchyma were the main abnormalities on MRI . Enhancing masses were not detected . These cases may represent two different reactions of the immunocompromised hosts to infection with C . neoformans: widening of the perivascular spaces as a correlate of the more typical meningeal infection and enhancing parenchymal lesions as a sign of further invasion from the CSF spaces . Enhancement of cryptococcomas, indicating an inflammatory response in the surrounding brain, is not typical in patients with impairment of immune function.

J Med Assoc Thai, 1999 Jan, 82(1), 65 - 71
Serum cyrptococcal antigen: diagnostic value in the diagnosis of AIDS-related cryptococcal meningitis; Asawavichienjinda T et al.; RATIONALE: The incidences of HIV-AIDS patients with opportunistic infections of the central nervous system are increasing . Of these, cryptococcal meningitis is the most important and serious . A simple method for the diagnosis of cryptococcal meningitis is needed despite its variable clinical features and the lack of a capacity in most health facilities in Thailand to exclude it from other diseases especially mass lesions in the brain . OBJECTIVE: To identify the capability and cut off point of serum cryptococcal antigen for diagnosis and screening of cryptococcal meningitis in HIV-AIDS patients . METHODS: One hundred consecutive cases of HIV-AIDS patients suspected of having central nervous system infections were prospectively recruited for the study . The serum of all patients were examined for cryptococcal antigen by latex agglutination test, the Pastorex Cryptococcus manufactured by Sanofi Diagnostic Pasteur, France . If a test was positive, the serum dilution was carried out using 10-fold serial dilution . Every patient went through pre-defined standard investigations to derive at a definite diagnosis . The gold standard for diagnosis of cryptococcal meningitis was the presence of encapsulated yeast forms in the cerebrospinal fluid or a positive culture for cryptococcal neoformans from the cerebrospinal fluid . RESULTS: Of 100 patients enrolled in this study, 58 patients had cryptococcal meningitis and serum cryptococcal antigen was detectable in 60 patients . If the cut-off point for a positive test was when the serum cryptococcal antigen titer was more than zero, then, the sensitivity of the test was 91.4 per cent, the specificity was 83.3 per cent, likelihood ratio if test positive (LR+) was 5.47, likelihood ratio if test negative (LR-) was 0.1, false positive was 16.7 per cent, false negative was 8.6 per cent . CONCLUSION: We conclude that serum cryptococcal antigen is a simple and rapid screening method for diagnosis of cryptococcal meningitis.

Mycoses, 1998, 41 Suppl 2, 81 - 5
{Incidence of endomycoses in the autopsy material of the Berlin Charité Hospital}; Tietz HJ et al.; From 1970 to 1993, a total of 93 endomf1p4es confirmed by post-mortem examination was diagnosed in the autopsy material of the Berlin Charite, a large hospital with an average of 1,500 hospital beds and maximum medical care . These comprised 51 candidoses (54.8%), 24 aspergilloses (36.6%), five cryptococcoses (5.4%), one zygomycosis, 1 trichosporosis and one coccidioidomycoses . This corresponded to 0.7% of the 13,375 decreased persons autopsied during this period . The frequency of autopsy was 85.3% . In 3,770 cases (2,418 adults and 1,352 children), brain dissection was performed . An adequate clinical putative diagnosis was made in only six out of 28 patients (18 adults, 10 children) with histologically confirmed cerebral mycosis {11 candidoses (39.3%), 10 aspergilloses (35.7%), five cryptococcoses (17.9%), one trichosporosis and one coccidioidomycosis} . About 80% of the mycoses of the CNS thus remained undetected while the patients were alive . Against the background of the continuing reduction in the frequency of autopsy in the Federal Republic of Germany, the observations made in the present paper underscore the demand for improved efficiency of mycological in-vivo diagnoses in the hospital and laboratory.

Infect Immun, 1999 Apr, 67(4), 1812 - 20
Cryptococcus neoformans differential gene expression detected in vitro and in vivo with green fluorescent protein; del Poeta M et al.; Synthetic green fluorescent protein (GFP) was used as a reporter to detect differential gene expression in the pathogenic fungus Cryptococcus neoformans . Promoters from the C . neoformans actin, GAL7, or mating-type alpha pheromone (MFalpha1) genes were fused to GFP, and the resulting reporter genes were used to assess gene expression in serotype A C . neoformans . Yeast cells containing an integrated pACT::GFP construct demonstrated that the actin promoter was expressed during vegetative growth on yeast extract-peptone-dextrose medium . In contrast, yeast cells containing the inducible GAL7::GFP or MFalpha1::GFP reporter genes expressed significant GFP activity only during growth on galactose medium or V-8 agar, respectively . These findings demonstrated that the GAL7 and MFalpha1 promoters from a serotype D C . neoformans strain function when introduced into a serotype A strain . Because the MFalpha1 promoter is induced by nutrient deprivation and the MATalpha locus containing the MFalpha1 gene has been linked with virulence, yeast cells containing the pMFalpha1::GFP reporter gene were analyzed for GFP expression in the central nervous system (CNS) of immunosuppressed rabbits . In fact, significant GFP expression from the MFalpha1::GFP reporter gene was detected after the first week of a CNS infection . These findings suggest that there are temporal, host-specific cues that regulate gene expression during infection and that the MFalpha1 gene is induced during the proliferative stage of a CNS infection . In conclusion, GFP can be used as an effective and sensitive reporter to monitor specific C . neoformans gene expression in vitro, and GFP reporter constructs can be used as an approach to identify a novel gene(s) or to characterize known genes whose expression is regulated during infection.

Chest, 1999 Mar, 115(3), 734 - 40
Pulmonary cryptococcosis in patients without HIV infection; Aberg JA et al.; PURPOSE: To further elucidate the diagnostic and therapeutic approaches to patients with pulmonary cryptococcosis who are not HIV-infected . SUBJECTS: All of the patients without HIV infection who received care at two Midwest hospitals between January 1986 and February 1996 and had a respiratory isolate of Cryptococcus neoformans . METHODS: The medical records of the study patients were reviewed for demographic data, host immune status, respiratory symptoms, diagnostic studies, treatment, and follow-up . RESULTS: Forty-two patient presentations comprised the overall study group . Thirty-six patients (85.7%) had no evidence of dissemination, and six patients (14.3%) had disseminated disease . Seven of the 36 patient presentations were definitive pulmonary cryptococcosis, 15 were presumptive disease, and 14 were colonization with C neoformans . Neither the baseline demographic parameters nor the immune status appeared to discriminate the patients with disease from the patients with colonization . A serum cryptococcal antigen (sCRAG) was positive for 7 of 18 patients, 3 of whom were proven by culture to have a disseminated infection . A negative sCRAG was observed in 11 patients, one of whom had proven dissemination . Fifteen patients underwent a lumbar puncture as part of their evaluation, and cryptococcal meningitis was diagnosed in three of these patients, all of whom had positive blood cultures for C neoformans . The majority of the patients did not receive antifungal therapy . CONCLUSION: In the majority of the patients, the lung appeared to be the sole organ involved, and a workup for systemic infection was rarely helpful . A positive sCRAG was not specific for dissemination . Antifungal therapy should be reserved for symptomatic patients, for patients with a positive sCRAG, and for patients with underlying immunosuppression.

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