Background Surface-enhanced laser desorption ionization time-of-flight mass spectrometry (SELDI-TOF-MS) was applied to analyze the protein profiles in both somatic and metabolic extracts of Aspergillus species. The study was carried out on some Aspergillus species within the Fumigati section ( Aspergillus fumigatus wild-types and natural abnormally pigmented mutants, and Aspergillus lentulus ). The aim was to validate whether mass spectrometry protein profiles can be used as specific signatures to discriminate different Aspergillus species or even mutants within the same species. Results The growth conditions and the SELDI-TOF parameters were determined to generate characteristic protein profiles of somatic and metabolic extracts of Aspergillus fumigatus strains using five different ProteinChips ® , eight growth conditions combining two temperatures, two media and two oxygenation conditions. Nine strains were investigated: three wild-types and four natural abnormally pigmented mutant strains of A. fumigatus and two strains of A. lentulus . A total of 242 fungal extracts were prepared. The spectra obtained are protein signatures linked to the physiological states of fungal strains depending on culture conditions. The best resolutions were obtained using the chromatographic surfaces CM10, NP20 and H50 with fractions of fungi grown on modified Sabouraud medium at 37°C in static condition. Under these conditions, the SELDI-TOF analysis allowed A. fumigatus and A. lentulus strains to be grouped into distinct clusters. Conclusions SELDI-TOF analysis distinguishes A. fumigatus from A. lentulus strains and moreover, permits separate clusters of natural abnormally pigmented A. fumigatus strains to be obtained. In addition, this methodology allowed us to point out fungal components specifically produced by a wild-type strain or natural mutants. It offers attractive potential for further studies of the Aspergillus biology or pathogenesis.

Phagocytes play a central role in immune defense. Their dysfunction predisposes to infections. This study determined the expression level of nine receptors involved in Aspergillus immune response as well as the values of phagocytosis and production of radical oxygen species after Aspergillus stimulation, in a healthy adult population. The expression values of the CD11b, CD11c, CD14, CD18, CD35, CD181, CD182, CD282 and CD284 receptors on peripheral human monocytes and granulocytes was established. A heterogenous expression of the CD282 on granulocytes was observed as CD181, CD182 and CD284 on monocytes. Similarly, we observed considerable variation in the expression of these receptors over time. Only CD282 on granulocytes varied with sex. No variation with age was observed. Adherence of Aspergillus conidia to phagocytes was dependent of individual, sex, age and time. A better characterization of these innate immunity parameters is necessary to develop in the future an immunologic surveillance strategy for transplant recipients.

Efficient monitoring of surfaces for spores of filamentous fungi is essential for detecting minor contamination even when air samples test negative for fungi. This study evaluates and compares a pad prepared using a dusting cloth with Rodac contact plates and humidified swabs for detecting mycological contamination, and concludes that the new method is superior and cheaper.

ABSTRACT Allergic bronchopulmonary aspergillosis (ABPA) is a frequent complication in cystic fibrosis patients. The diagnosis remains difficult and requires a combination of clinical, radiological, biological, and mycological criteria. The aim of this study was to analyze the added value of two recombinant antigens, rAspf4 and rAspf6, associated with the detection of specific IgG; precipitins; total IgE; and Aspergillus fumigatus in sputum for the diagnosis of ABPA. In a retrospective study, we determined the specific IgE responses to these recombinants in 133 sera of 65 cystic fibrosis patients. We selected an average of five serum samples from each of the 17 patients with ABPA (13 proven and 4 probable ABPA) and from 3 patients with Aspergillus bronchitis and rhinosinusitis. One serum sample for the 45 patients without ABPA was tested. The sensitivity of specific IgE detection against rAspf4 calculated per patient (92.3%) was significantly higher (P < 0.05) than that of rAspf6 (53.8%). When rAspf4 IgE detection was associated with anti-Aspergillus IgG enzyme-linked immunosorbent assay (ELISA) and precipitin detection, the sensitivity rose to 100%. The specificities of rAspf4 and rAspf6 IgE detection were 93.7% and 91.6%, respectively. Other diagnostic criteria had slightly lower specificities (87.5% for anti-Aspergillus IgG ELISA, 89.6% for precipitins, 84.4% for total IgE, and 85.0% for positive A. fumigatus culture in sputum). In conclusion, this retrospective study showed the relevance of rAspf4 IgE detection, in combination with other biological markers (Aspergillus IgG ELISA, precipitins, and total IgE), for improving the biological diagnosis of ABPA.

Although data show the importance of type I interferons (IFNs) in the regulation of the innate and adaptive immunity elicited in response to viral, bacterial and parasitic infections, the functional activities of these cytokines during fungal infections are poorly understood. We examined here the impact of IFN-β on the response of human monocyte-derived dendritic cells (DCs) infected in vitro with Aspergillus fumigatus. Having found that A. fumigatus-infected DCs do not express IFN-β, we evaluated the effect of the exogenous addition of IFN-β on the maturation of human DCs induced by the infection with A. fumigatus conidia. Although the phagocytosis of the fungus was not affected by IFN-β treatment, the expression of CD86 and CD83 induced upon A. fumigatus challenge was enhanced in IFN-β-conditioned DCs, which also showed an increased expression of IL-27 and IL-12p70, members of IL-12 family. Through these modifications, IFN-β improved the capacity of DCs to promote an anti-Aspergillus T helper type 1 response, as evaluated by mixed leucocyte reaction, which plays a crucial role in the control of invasive aspergillosis. Our results identified a novel effect of IFN-β on anti-Aspergillus immune responses which, in turn, might open new perspectives on the use of IFN-β in immunotherapy for fungal infections aimed at enhancing the immunological functions of DCs.

Background: Nosocomial invasive filamentous fungi infections could result from inhalation of filamentous fungi conidia present in hospital environment.Methods: The environmental fungal flora in 3 different hospital wards with similar air conditioning was prospectively studied during 30 months and compared to internal (presence of agranulocytosis patient, behavioral practices, activity, cleaning work) and outdoor factors (meteorologic data, outdoor fungi). The general preventive measures differed from one unit to another.Results: The hematology wards with filamentous fungi preventive measures were significantly less contaminated than a conventional ward without specific measures. internal and outdoor factors influenced the level of fungal flora. However, the influence of internal factors was greater in the conventional ward than in hematology wards. The variation of flora in the hospital environment was seasonal, and the level of this contamination in each ward was influenced by the meteorology However, outdoor factors more readily explain the variations of fungal load in hematology than in the conventional ward.Conclusion: This study highlights that specific preventive measures participate significantly in the control of the filamentous fungal flora intensity due to internal factors but not those due to outdoor factors, stressing the importance of high-efficiency particulate air filtration in high-risk units.

Air treatment with a mobile Plasmair air-decontamination unit significantly reduces the fungal spore load in hematology wards. We report that this system used at a low aspiration flow does not perform total biodecontamination against filamentous fungi. moreover, the filamentous fungus load remaining in rooms equipped with this mobile air-decontamination unit is lowest in wards in which other preventive measures against nosocomial filamentous fungal infections are implemented. Copyright (C) 2009 by the Association for Professionals in Infection Control and Epidemiology, Inc. (Am J Infect Control 2009;37:680-2.)

ABSTRACT In order to shed light on its basic biology, we initiated a population genetic analysis of Candida glabrata , an emerging pathogenic yeast with no sexual stage yet recognized. A worldwide collection of clinical strains was subjected to analysis using variable number of tandem repeats (VNTR) at nine loci. The clustering of strains obtained with this method was congruent with that obtained using sequence polymorphism of the NMT1 gene, a locus previously proposed for lineage assignment. Linkage disequilibrium supported the hypothesis of a mainly clonal reproduction. No heterozygous diploid genotype was found. Minimum-spanning tree analysis of VNTR data revealed clonal expansions and associated genotypic diversification. Mating type analysis revealed that 80% of the strains examined are MAT a and 20% MAT α and that the two alleles are not evenly distributed. The MAT a genotype dominated within large clonal groups that contained only one or a few MAT α types. In contrast, two groups were dominated by MAT α strains. Our data are consistent with rare independent mating type switching events occurring preferentially from type a to α, although the alternative possibility of selection favoring type a isolates cannot be excluded.

We describe 6 cases of severe filamentous fungal infections after widespread tissue damage due to traumatic injury in previously healthy people. Additionally, we report 69 cases from an exhaustive 20-y review of the literature to investigate the epidemiological and clinical features, the prognosis and the therapeutic management of these post-traumatic severe filamentous fungal infections. Traffic (41%) and farm accidents (25%) were the main causes of injury, which involved either the limbs only (41%) or multiple sites (41%). Necrosis was the main symptom (60%) and Mucorales (72%) and Aspergillus (11%) were the 2 most frequent fungi causing infection. These infections required substantial surgical debridement or amputation (96%) associated with aggressive antifungal therapy (81%), depending on the responsible fungi. This study underlines the need for early, repeated and systematic mycological wound samples to guide and adapt surgical and antifungal management in these filamentous fungal infections.

Innate immunity is the major host defense against invasive aspergillosis. To determine whether the collectin mannan-binding lectin (MBL) is involved in the initial protective immunity through complement activation against opportunistic fungal infections caused by Aspergillus, we performed in vitro studies on 29 different strains of Aspergillus conidia from five different species. Incubation of Aspergillus conidia in human normal serum leads to activation of the alternative pathway, whereas neither the classical nor the lectin pathways through C4 and C2 cleavage are activated. Complement response to conidia was investigated using a MBL-deficient serum and reconstitution experiments were conducted with MBL/MASPs complexes. We found that MBL can directly support C3 activation by a C2 bypass mechanism. Finally, a stronger activation of the alternative pathway was observed for the clinical strains isolated from patients with invasive aspergillosis, compared with the environmental strains.

Given the role played by chemokines in the selective homing of immune cells, we sought to characterize the profile of chemokines produced by human dendritic cells (DC) following in vitro Aspergillus fumigatus infection and their ability to recruit cells involved in the antifungal defense. At the onset of A. fumigatus infection, DC released elevated amounts of CXCL8 that promote the migration of polymorphonuclear cells (PMN). Moreover, soluble factors released from A. fumigatus-infected DC increased also the surface expression of two activation markers, CD11b and CD18, on PMN. A. fumigatus infection resulted also in CCL3, CCL4, CXCL10 and CCL20 productions that induce the migration of effector memory Th1 cells. Moreover, the late expression of CCL19 suggests that A. fumigatus-infected DC could be implicated in the migration of CCR7+ naïve T cells and mature DC in lymph nodes. Together these results suggested the involvement of human DC in the regulation of innate and adaptive immunity against A. fumigatus through the recruitment of cells active in the fungal destruction.

Despite the widespread use of antifungals for prophylaxis, Candida bloodstream infection (BSI) remains the most frequent life-threatening fungal disease. From an analysis of multi-institutional surveys of Candida BSIs performed in Europe, including the large prospective survey by the European Confederation of Medical Mycology (2089 episodes from seven countries), a limited role of species with decreased susceptibility to azoles in causing BSIs and a low proportion of antifungal resistance was evident. Large prospective epidemiological surveys using common databases are needed to monitor trends in incidence and changes in species distribution, to identify new at-risk patients and to evaluate the impact of the introduction into the market of new antifungal agents.

Human cytomegalovirus (HCMV) infection is associated with an increased susceptibility to opportunistic infections. Although the subversion of adaptive immune responses has been extensively studied, the consequences of HCMV infection on natural immune responses are not well documented. A striking selective downmodulation of CD11b/CD18 (CR3) or CD11c/CD18 (CR4) was found upon HCMV infection, on two models, the monocytic THP-1 cell line and monocyte- derived macrophages. HCMV-infected macrophages have an altered adhesion/phagocytic capacity to Candida albicans, a pathogen responsible for some opportunistic infections in immunocompromised patients. These results suggest a new mechanism implicated in the augmentation of opportunistic infections in HCMV patients.

A recent study by Horvath et al. (5) assessed the use of BACTEC Plus Aerobic/F and Anaerobic/F blood culture (BC) bottles with the BACTEC 9240 system to detect yeasts in a simulated-candidemia model. The authors concluded that further research was needed to optimize the recovery of Candida spp. from this automated BC system because of the high rate of false negatives and the long time to detection (TTD), especially for Candida glabrata. We wish to comment on this study and report our own experience.

Human cryptosporidiosis : an emerging parasitic disease of increasing importance in public health. Cryptosporidiosis in human is an emerging parasitosis due to an intestinal and opportunistic protozoan : Cryptosporidium parvum. This parasite was described 100 years ago in calves and lambs. This zoonosis had emerged as a global public health problem in industrial countries for essentially two reasons. One is linked to its potentially life threatening evolution which remains practiccaly incurable, in severely immunocompromised patients (AIDS patients, transplant patients, young children with immature immunological system...). The other is the important warterbome outbreaks in industrial countries. Clinical forms of cryptosporidiosis range from asymptomatic to acute intestinal illness in immunocompetent host. In immunocompromised host, chronic or severe diarrhoeal diseases increase the morbidity and mortality. Moreover extra intestinal dissemination of parasites can accured (cholangitis) and pulmonary cryptosporidiosis are also frequently discribed. This parasitosis is also caracterized by the lack of efficient therapy and insuffisant prophylactic measures. Numerical standard and legal regulation of Cryptosporidium was proposed. Furthermore the oocyts are extremelly resistant to conventional disinfectants. The control of water treatment is necessary to determine the efficiency of prophylactic measures. The significance of different Cryptosporidium parvum isolates could suggested the existence of different transmission cycles in human and merits futher investigations. The virulent potential of Cryptosporidium strains as well as individual susceptibility need further works to a better undestanding of epidemiological risk.

To analyse if hydrocortisone could modify the in vitro susceptibility of Aspergillus fumigatus to antifungal drugs, we developed a procedure to test the susceptibility of an A. fumigatus strain to amphotericin B and itraconazole, grown in the presence and in the absence of hydrocortisone. Conidia were germinated in the presence or the absence of hydrocortisone in Czapek medium without antifungal drug. A dilution of these cultures (5x10(3) conidia ml(-1)) was spread onto Czapek-agarose plates containing both antifungal drug and hydrocortisone. The cfu per plate were enumerated and compared. A therapeutic concentration of hydrocortisone induced a significant increase in the susceptibility to itraconazole. Conversely, the susceptibility to amphotericin B was not significantly modified when this antifungal drug was associated with hydrocortisone.

In order to evaluate bacterial and fungal contamination, the authors used a qualitative and quantitative procedure and investigated 47 air conditioning and humidifying units.

Sir—In their recent article, Piscitelli et al. [1] advise that " patients receiving saqui-navir as their sole protease inhibitor should avoid using garlic supplements " [1, p. 238]. In their study, healthy volunteers who received the antiretroviral drug sa-quinavir and a garlic supplement exhibited a significant decline in plasma levels of saquinavir. As one reason for the decline in systemic levels of saquinavir, the authors suggest that the effect " may be caused by the induction of CYP450 [cy-tochrome P450] in the gut mucosa " by garlic supplements [1, p. 237]. This warning against taking garlic supplements , stated in the article and publicized in the media, is based on results with a single supplement, GarliPure Maximum Allicin Formula (Natrol), that is standardized with allicin, an unstable compound that is known to convert to polysulfides that induce the production of cytochrome P450. The study and its conclusions do not address the availability of garlic supplements that are not standardized with allicin. For example, Kyolic Aged Garlic Extract (Wakunaga), which is standardized by water soluble S-allyl cysteine, is devoid of allicin and does not induce the production of cytochrome P450. Thus, a limited conclusion by Piscitelli and colleagues would have been more prudent—namely, that patients receiving saquinavir as their sole protease inhibitor should avoid taking the allicin-standardized garlic supplement they studied. Although additional clinical research is needed, use of a supplement that is not standardized with allicin, such as Kyolic Aged Garlic Extract, may be a safe option for people being treated with saquinavir. Reply Sir—We agree with Dr. Borek [1] that, despite having data only on the interaction between a single garlic preparation and sa-quinavir, we drew a generalized conclusion regarding the need for patients to use caution if they combine garlic supplements with saquinavir when using that drug as the sole protease inhibitor [2]. Because we have no information to suggest which constituent (or excipient) in the garlic formulation is responsible for the drug interaction, we cannot speculate about the effects of other commercial products or dietary garlic on the phar-macokinetics of saquinavir or relate our findings to allicin concentrations. We provided data on the allicin (and allin) content of the supplement we studied solely because we considered the verification of product content to be important. Since publication of our article, we have had a study brought to our attention in which garlic supplements were tested for drug release …