Corticosteroid treatment is associated with increased filamentous fungal burden in allergic fungal disease

Background Allergic diseases caused by fungi are common. The best understood conditions are allergic bronchopulmonaryaspergillosis (ABPA) and severe asthma with fungal sensitisation (SAFS). Our knowledge of the fungal microbiome (mycobiome) is limited to a few studies involving healthy individuals, asthmatics and smokers. No study has yet examined the mycobiome in fungal lung disease. Objectives The main aim of this study was to determine the mycobiome in lungs of individuals with well characterised fungal disease. A secondary objective was to determine possible effects of treatment on the mycobiome. Methods After bronchoscopy, ITS1 DNA was amplified and sequenced and fungal load determined by RT-PCR. Clinical and treatment variables were correlated with the main species identified. ABPA (n=16), SAFS (n=16), severe asthma not sensitised to fungi, (n=9), mild asthma patients(n=7) and 10 healthy controls were studied. Results The mycobiome was highly varied with severe asthmatics carrying higher loads of fungus. Healthy individuals had low fungal loads, mostly poorly characterised Malasezziales.The most common fungus in asthmatics was Aspergillus fumigatus complex and this taxon accounted for the increased burden of fungus in the high level samples. Corticosteroid treatment was significantly associated with increased fungal load (p<0.01). Conclusions The mycobiome is highly variable. Highest loads of fungus are observed in severe asthmatics and the most common fungus is Aspergillus fumigatus complex. Individuals receiving steroid therapy had significantly higher levels of Aspergillus and total fungus in their BAL

A spatial approach for the epidemiology of antibiotic use and resistance in community-based studies: the emergence of urban clusters of Escherichia coli quinolone resistance in Sao Paulo, Brasil

Copyright © Kiffer et al; licensee BioMed Central Ltd. 2011 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.Background Population antimicrobial use may influence resistance emergence. Resistance is an ecological phenomenon due to potential transmissibility. We investigated spatial and temporal patterns of ciprofloxacin (CIP) population consumption related to E. coli resistance emergence and dissemination in a major Brazilian city. A total of 4,372 urinary tract infection E. coli cases, with 723 CIP resistant, were identified in 2002 from two outpatient centres. Cases were address geocoded in a digital map. Raw CIP consumption data was transformed into usage density in DDDs by CIP selling points influence zones determination. A stochastic model coupled with a Geographical Information System was applied for relating resistance and usage density and for detecting city areas of high/low resistance risk. Results E. coli CIP resistant cluster emergence was detected and significantly related to usage density at a level of 5 to 9 CIP DDDs. There were clustered hot-spots and a significant global spatial variation in the residual resistance risk after allowing for usage density. Conclusions There were clustered hot-spots and a significant global spatial variation in the residual resistance risk after allowing for usage density. The usage density of 5-9 CIP DDDs per 1,000 inhabitants within the same influence zone was the resistance triggering level. This level led to E. coli resistance clustering, proving that individual resistance emergence and dissemination was affected by antimicrobial population consumption

Identification of a Putative Crf Splice Variant and Generation of Recombinant Antibodies for the Specific Detection of Aspergillus fumigatus

BACKGROUND: Aspergillus fumigatus is a common airborne fungal pathogen for humans. It frequently causes an invasive aspergillosis (IA) in immunocompromised patients with poor prognosis. Potent antifungal drugs are very expensive and cause serious adverse effects. Their correct application requires an early and specific diagnosis of IA, which is still not properly achievable. This work aims to a specific detection of A. fumigatus by immunofluorescence and the generation of recombinant antibodies for the detection of A. fumigatus by ELISA. RESULTS: The A. fumigatus antigen Crf2 was isolated from a human patient with proven IA. It is a novel variant of a group of surface proteins (Crf1, Asp f9, Asp f16) which belong to the glycosylhydrolase family. Single chain fragment variables (scFvs) were obtained by phage display from a human naive antibody gene library and an immune antibody gene library generated from a macaque immunized with recombinant Crf2. Two different selection strategies were performed and shown to influence the selection of scFvs recognizing the Crf2 antigen in its native conformation. Using these antibodies, Crf2 was localized in growing hyphae of A. fumigatus but not in spores. In addition, the antibodies allowed differentiation between A. fumigatus and related Aspergillus species or Candida albicans by immunofluorescence microscopy. The scFv antibody clones were further characterized for their affinity, the nature of their epitope, their serum stability and their detection limit of Crf2 in human serum. CONCLUSION: Crf2 and the corresponding recombinant antibodies offer a novel approach for the early diagnostics of IA caused by A. fumigatus

Habitat use at fine spatial scale: how does patch clustering criteria explain the use of meadows by red deer ?

Large mammalian herbivores are keystone species in different ecosystems. To mediate the effects of large mammalian herbivores on ecosystems, it is crucial to understand their habitat selection pattern. At finer scales, herbivore patch selection depends strongly on plant community traits and therefore its understanding is constrained by patch definition criteria. Our aim was to assess which criteria for patch definition best explained use of meadows by wild, free-ranging, red deer (Cervus elaphus) in a study area in Northeast Portugal. We used two clustering criteria types based on floristic composition and gross forage classes, respectively. For the floristic criteria, phytosociological approach was used to classify plant communities, and its objectivity evaluated with a mathematical clustering of the floristic relevés. Cover of dominant plant species was tested as a proxy for the phytosociological method. For the gross forage classes, the graminoids/forbs ratio and the percentage cover of legumes were used. For assessing deer relative use of meadows we used faecal accumulation rates. Patches clustered according to floristic classification better explained selection of patches by deer. Plant community classifications based on phytosociology, or proxies of this, used for characterizing meadow patches resulted useful to understand herbivore selection pattern at fine scales and thus potentially suitable to assist wildlife management decisions

Intelligent Interfaces to Empower People with Disabilities

Severe motion impairments can result from non-progressive disorders, such as cerebral palsy, or degenerative neurological diseases, such as Amyotrophic Lateral Sclerosis (ALS), Multiple Sclerosis (MS), or muscular dystrophy (MD). They can be due to traumatic brain injuries, for example, due to a traffic accident, or to brainste

Clinical performance of FXG: RESP (Asp+) assay for Pneumocystis jirovecii on respiratory specimens

Objectives: Pneumocystis pneumonia continues to be a common infection in HIV patients and also occurs in other immunocompromised patients. Early diagnosis is known to improve outcome, and specifically, exclusion of the diagnosis, reduces the need for toxic empirical high dose cotrimoxazole therapy. Real-Time PCR offers the prospect of faster and highly sensitive detection of P. jirovecii. FXG : RESP (Asp +) [Myconostica, UK] is a test kit that detects both P. jirovecii and Aspergillus spp., utilising molecular beacons. In this report we focus on the clinical performance for P. jirovecii. Methods: The FXG : RESP (Asp +) real-time PCR kit utilises the large subunit mtRNA gene as a target to detect P. jirovecii and the assay is highly sensitive, being able to detect 6 target copies. The assay appears to be specific for Pneumocystis spp, with the possible exception of Fusarium solani. It was tested blindly on 196 BAL samples, collected from 4 European hospitals. All results were compared to microscopy, usually Calcofluor or Gomori methanamine silver stains, performed shortly after the sample was collected, and in non-AIDS patients whether patient was treated for PCP. Most HIV/AIDS samples had been previously extracted and stored frozen as DNA; the remainder were extracted with the MycXtra™ fungal DNA extraction kit, having been stored frozen unprocessed. Results: The kit contains an internal amplification control to detect potential inhibition of the PCR reaction and 6 (3%) of the clinical samples showed evidence of inhibition. These results were excluded from analysis, although 5 were positive by both microscopy and the FXG : RESP (Asp +) assay, and would be reported clinically. 42 samples were from HIV/AIDS patients and 148 from other patients, mostly with leukaemia. With respect to Pneumocystis detection, the FXG : RESP (Asp +) assay had good performance with a sensitivity of 97.4%, specificity of 92.9%, positive and negative predictive values of 90.4% and 98.1%. 8 samples had negative microscopy but very high FXG assay signals, suggesting that the microscopy was falsely negative, as reported in prior literature. These were reported as false positives Conclusions: The clinical performance of the FXG : RESP (Asp +) assay for the diagnosis of Pneumocystis pneumonia is superb. Overall the speed of detection and sensitivity of the FXG : RESP (Asp +) assay should bring substantial clinical benefits. Prospective and supportive clinical trials are ongoing.status: publishe