JoVE Visualize What is visualize?
Stop Reading. Start Watching.
Advanced Search
Stop Reading. Start Watching.
Regular Search
Find video protocols related to scientific articles indexed in Pubmed.
The clinical spectrum of pulmonary aspergillosis.
Thorax
PUBLISHED: 10-31-2014
Show Abstract
Hide Abstract
The clinical presentation of Aspergillus lung disease is determined by the interaction between fungus and host. Invasive aspergillosis develops in severely immunocompromised patients, including those with neutropenia, and increasingly in the non-neutropenic host, including lung transplant recipients, the critically ill patients and patients on steroids. A high index of suspicion is required in patients without the classical risk factors as early presentation is usually silent and non-specific, pyrexia uncommon and timely treatment is crucial for survival. Invasive aspergillosis has also been diagnosed in normal hosts after massive exposure to fungal spores. Chronic pulmonary aspergillosis affects patients without obvious immune compromise, but with an underlying lung condition such as COPD or sarcoidosis, prior or concurrent TB or non-tuberculous mycobacterial disease. Aspergillus bronchitis may be responsible for persistent respiratory symptoms in patients with Aspergillus detected repeatedly in sputum without evidence of parenchymal Aspergillus disease, especially in patients with bronchiectasis and cystic fibrosis. Allergic bronchopulmonary aspergillosis affects patients with asthma and cystic fibrosis, and is important to recognise as permanent lung or airways damage may accrue if untreated. Changes in the classification of Aspergillus allergic lung disease have been proposed recently. Cases of extrinsic allergic alveolitis and chronic pulmonary aspergillosis have been observed after Aspergillus exposure. Asymptomatic colonisation of the respiratory tract needs close monitoring as it can lead to clinical disease especially with ongoing immunosuppression. The various syndromes should be viewed as a semicontinuous spectrum of disease and one form may evolve into another depending on the degree of ongoing immunosuppression.
Related JoVE Video
Name changes in medically important fungi and their implications for clinical practice.
J. Clin. Microbiol.
PUBLISHED: 10-10-2014
Show Abstract
Hide Abstract
Recent changes in the Fungal Code of Nomenclature and developments in molecular phylogeny are about to lead to dramatic changes in the naming of medically important moulds and yeasts. In this article, we present a widely supported and simple proposal to prevent unnecessary nomenclatural instability.
Related JoVE Video
Efficacy and safety of nebulised amphotericin B (NAB) in severe asthma with fungal sensitisation (SAFS) and allergic bronchopulmonary aspergillosis (ABPA).
J Asthma
PUBLISHED: 09-23-2014
Show Abstract
Hide Abstract
Abstract Background and rationale: Antifungal therapy for severe asthma with fungal sensitisation (SAFS) and allergic bronchopulmonary aspergillosis (ABPA) remains poorly studied. We assessed the efficacy and safety of NAB as second and third line therapy in SAFS and ABPA. Methods: 21 adult asthmatics with SAFS (n?=?11) and ABPA (n?=?10) who had either failed itraconazole (n?=?8), voriconazole proceeded by itraconazole (n?=?5) or developed adverse events (AEs) to either agent (n?=?7) were treated with 10mg of NAB (Fungizone) twice daily. We audited clinical and immunological response, using the Asthma Quality of Life Questionnaire (AQLQ-J) scores, asthma control, FEV1, healthcare utilisation and IgE. Patients were followed up for 12 months. Results: Twenty-one patients were treated (SAFS, n?=?11) and (ABPA, n?=?10), M: F?=?8:12, median age 65 years (range, 24-78). The median duration of therapy was 30 days (0-1825). Clinical benefit was observed in three (14.3 %) in which overall mean AQLQ-J score improved by?+?2.9, mean FEV1 improved by 0.5 L and there was improvement in overall asthma control. Seven (33%) failed initial dose (bronchospasm). Eleven (52.4%) discontinued within 12 months of therapy due to delayed bronchospasm (n?=?3, within 4 weeks), equipment problems (n?=?2, within 4 weeks) and lack of clinical benefit (n?=?4, within 16 weeks). Conclusion: Our data suggest that the overall efficacy of NAB in this group of patients is poor and associated with bronchospasm. However, the excellent response in 3 patients, suggest it may be considered when other alternatives have been exhausted. Overcoming the initial bronchospasm may improve tolerability.
Related JoVE Video
Elevated Levels of the Neutrophil Chemoattractant Pro-Platelet Basic Protein in Macrophages From Individuals With Chronic and Allergic Aspergillosis.
J. Infect. Dis.
PUBLISHED: 09-05-2014
Show Abstract
Hide Abstract
?Aspergillus fumigatus causes chronic cavitary pulmonary aspergillosis (CCPA) and allergic bronchopulmonary aspergillosis (ABPA) in overtly immunocompetent and atopic individuals, respectively. Disease mechanisms are poorly understood but may be related to increased neutrophil presence and activation. Pro-platelet basic protein (PPBP) is a potent neutrophil chemoattractant and activator whose expression is repressed by interleukin 10 (IL-10).
Related JoVE Video
Emerging novel and antimicrobial-resistant respiratory tract infections: new drug development and therapeutic options.
Lancet Infect Dis
PUBLISHED: 09-01-2014
Show Abstract
Hide Abstract
The emergence and spread of antimicrobial-resistant bacterial, viral, and fungal pathogens for which diminishing treatment options are available is of major global concern. New viral respiratory tract infections with epidemic potential, such as severe acute respiratory syndrome, swine-origin influenza A H1N1, and Middle East respiratory syndrome coronavirus infection, require development of new antiviral agents. The substantial rise in the global numbers of patients with respiratory tract infections caused by pan-antibiotic-resistant Gram-positive and Gram-negative bacteria, multidrug-resistant Mycobacterium tuberculosis, and multiazole-resistant fungi has focused attention on investments into development of new drugs and treatment regimens. Successful treatment outcomes for patients with respiratory tract infections across all health-care settings will necessitate rapid, precise diagnosis and more effective and pathogen-specific therapies. This Series paper describes the development and use of new antimicrobial agents and immune-based and host-directed therapies for a range of conventional and emerging viral, bacterial, and fungal causes of respiratory tract infections.
Related JoVE Video
What is the importance of classifying Aspergillus disease in cystic fibrosis patients?
Expert Rev Respir Med
PUBLISHED: 05-29-2014
Show Abstract
Hide Abstract
Aspergillus species are commonly isolated from lower respiratory tract samples of patients with cystic fibrosis (CF) and markers of immunological sensation to Aspergillus are frequently encountered in this group of patients; however, the contribution of Aspergillus to CF lung disease outside of the typical complications of ABPA and aspergilloma formation remains largely unclear. Patients with CF show discretely different responses to Aspergillus, though the underlying reasons for this variation are unknown. Recent work has begun to allow us to categorize patient responses to Aspergillus based upon molecular markers of infection and immune sensitization. Aspergillus sensitization and/or airway infection is associated with worse FEV1, in CF and other patients (asthma, chronic obstructive pulmonary disease, bronchiectasis). Classification of different clinical phenotypes of Aspergillus will enable future studies to determine the natural history of different manifestations of Aspergillus disease and evaluate the effects of intervention with antifungal therapy.
Related JoVE Video
Long-term stability at -20 °C of Aspergillus galactomannan in serum and bronchoalveolar lavage specimens.
J. Clin. Microbiol.
PUBLISHED: 04-09-2014
Show Abstract
Hide Abstract
Research to develop and validate novel methods for diagnosis of aspergillosis based on detection of galactomannan requires the use of clinical specimens that have been stored frozen. Data indicating that galactomannan remains stable when frozen are scant. The objective of this study was to determine the stability of galactomannan in clinical specimens stored at -20 °C that were positive in the Platelia Aspergillus enzyme immunoassay when initially tested. Prospective real-time testing of serum and bronchoalveolar lavage (BAL) fluid pools from positive and negative patient specimens showed no decline in galactomannan index (GMI) over 11 months at -20 °C and no development of positive reactions in the negative-control pool. Retrospective testing of positive specimens that had been stored at -20 °C for 5 years showed that 28 of 30 serum (n = 15) or BAL (n = 15) specimens remained positive. These findings support the use of frozen serum or BAL specimens stored for at least 5 years in evaluation of diagnostic tests based on detection of galactomannan.
Related JoVE Video
Posaconazole responsive cerebral aspergillosis in an immunocompetent adult.
J Clin Neurosci
PUBLISHED: 04-01-2014
Show Abstract
Hide Abstract
Cerebral aspergillosis is a rare manifestation of invasive aspergillosis that usually affects immunocompromised patients. There are few treatment options for recurrent disease and experiences with immunocompetent patients are lacking. We report the clinical course of an immunocompetent patient with recurrent cerebral aspergillosis, following initial treatment with burr hole aspiration and voriconazole, who showed remarkable response to posaconazole. The patient remains clinically well with no evidence of recurrence on MRI 7 years following diagnosis. To our knowledge this is the first reported experience with posaconazole in an immunocompetent patient with cerebral aspergillosis.
Related JoVE Video
Frequency of Pneumocystis jirovecii in sputum from HIV and TB patients in Namibia.
J Infect Dev Ctries
PUBLISHED: 03-13-2014
Show Abstract
Hide Abstract
The opportunistic fungus Pneumocystis jirovecii causes Pneumocystis pneumonia (PcP), which is a life-threatening infection in HIV/AIDS patients. The seemingly low prevalence of P. jirovecii pneumonia in sub-Saharan Africa has been a matter of great debate because many HIV/AIDS patients reside in this region. The lack of suitable diagnostic practices in this resource limited-region has been added to the uncertainty of PcP prevalence. Only a few studies have evaluated the utility of easily obtainable samples such as expectorated sputum for diagnosis of PcP. Thus, the aim of the current study was to evaluate the effectiveness of expectorated sputum for the routine diagnosis of PcP in a resource-limited sub-Saharan African setting.
Related JoVE Video
AIDS-related mycoses: the way forward.
Trends Microbiol.
PUBLISHED: 03-04-2014
Show Abstract
Hide Abstract
The contribution of fungal infections to the morbidity and mortality of HIV-infected individuals is largely unrecognized. A recent meeting highlighted several priorities that need to be urgently addressed, including improved epidemiological surveillance, increased availability of existing diagnostics and drugs, more training in the field of medical mycology, and better funding for research and provision of treatment, particularly in developing countries.
Related JoVE Video
Multi-country estimate of different manifestations of aspergillosis in cystic fibrosis.
PLoS ONE
PUBLISHED: 01-01-2014
Show Abstract
Hide Abstract
Aspergillus spp. can lead to allergic bronchopulmonary aspergillosis (ABPA), Aspergillus sensitisation and Aspergillus bronchitis in CF. The relative frequencies of these entities have recently been ascertained in a large UK adult CF cohort. We have used this data to estimate the burden of aspergillosis and ABPA cases in adult CF patients in 30 countries reporting CF. National and international CF registry data was accessed and assessed for completeness and age distribution. Published proportions of ABPA (17.7%), Aspergillus sensitisation (14.6%) and Aspergillus bronchitis (30%) in CF were applied to those >18 years and compared with notified ABPA cases. Of the 76,201 estimated CF patients worldwide (not including India), 37,714 were >18 years. The proportion of adults to children varied from 63% in Norway to 20% in Brazil. ABPA caseload in adults is anticipated to be 6,675 cases of which only 2,221 cases (33%) are currently recorded, indicating substantial underdiagnosis. The ABPA diagnosis rate compared with estimated rates varies by country from 101% (France) to 14.5% (Greece), although genetic variation could account for genuine differences compared with the UK. Aspergillus bronchitis is not currently recognised or recorded in CF registries but there are an anticipated 10,988 adult cases. Aspergillus sensitisation, associated with increased bronchiectasis and reduced FEV1, affects an anticipated 5,506 patients without ABPA or Aspergillus bronchitis. Together ABPA and Aspergillus bronchitis are estimated to affect 17,989 adults, 47.7% of the adult CF population. ABPA also occurs in children and teenagers and 984 cases were documented in registries. Diagnosed ABPA rates by age were available for the ECFS registry, USA, UK, Ireland, Belgium and Netherlands. The rate was <1% under 4 years, and increased throughout childhood and adolescence, with marked variation between countries. Newly published diagnostic criteria and methods should facilitate better recognition of aspergillosis in CF, allowing better CF disease control.
Related JoVE Video
Fungal allergy in asthma-state of the art and research needs.
Clin Transl Allergy
PUBLISHED: 01-01-2014
Show Abstract
Hide Abstract
Sensitization to fungi and long term or uncontrolled fungal infection are associated with poor control of asthma, the likelihood of more severe disease and complications such as bronchiectasis and chronic pulmonary aspergillosis. Modelling suggests that >6.5 million people have severe asthma with fungal sensitizations (SAFS), up to 50% of adult asthmatics attending secondary care have fungal sensitization, and an estimated 4.8 million adults have allergic bronchopulmonary aspergillosis (ABPA). There is much uncertainty about which fungi and fungal allergens are relevant to asthma, the natural history of sensitisation to fungi, if there is an exposure response relationship for fungal allergy, and the pathogenesis and frequency of exacerbations and complications. Genetic associations have been described but only weakly linked to phenotypes. The evidence base for most management strategies in ABPA, SAFS and related conditions is weak. Yet straightforward clinical practice guidelines for management are required. The role of environmental monitoring and optimal means of controlling disease to prevent disability and complications are not yet clear. In this paper we set out the key evidence supporting the role of fungal exposure, sensitisation and infection in asthmatics, what is understood about pathogenesis and natural history and identify the numerous areas for research studies.
Related JoVE Video
IgE-mediated immune responses and airway detection of Aspergillus and Candida in adult cystic fibrosis.
Chest
PUBLISHED: 08-30-2013
Show Abstract
Hide Abstract
The recovery of Aspergillus and Candida from the respiratory secretions of patients with cystic fibrosis (CF) is common. Their relationship to the development of allergic sensitization and effect on lung function has not been established. Improved techniques to detect these organisms are needed to increase knowledge of these effects.
Related JoVE Video
Long-term antifungal treatment improves health status in patients with chronic pulmonary aspergillosis: a longitudinal analysis.
Clin. Infect. Dis.
PUBLISHED: 06-20-2013
Show Abstract
Hide Abstract
?Chronic pulmonary aspergillosis (CPA) is an infectious disease that progressively destroys lung tissue. To date, no longitudinal data on the efficacy of antifungal treatment on health status in CPA patients exist.
Related JoVE Video
Validity and reliability of the St. Georges Respiratory Questionnaire in assessing health status in patients with chronic pulmonary aspergillosis.
Chest
PUBLISHED: 06-14-2013
Show Abstract
Hide Abstract
Chronic pulmonary aspergillosis (CPA) markedly reduces lung function through progressive lung destruction. To date, however, health status in patients with CPA has not been studied. This is due, in part, to a lack of adequately validated scales. The St. Georges Respiratory Questionnaire (SGRQ) is widely used for several chronic respiratory diseases, but not for CPA. We examined the reliability and validity of SGRQ in CPA.
Related JoVE Video
Results of surgery for chronic pulmonary Aspergillosis, optimal antifungal therapy and proposed high risk factors for recurrence - a National Centres experience.
J Cardiothorac Surg
PUBLISHED: 05-14-2013
Show Abstract
Hide Abstract
Surgery for pulmonary aspergillosis is infrequent and often challenging. Risk assessment is imprecise and new antifungals may ameliorate some surgical risks. We evaluated the medical and surgical management of these patients, including perioperative and postoperative antifungal therapy.
Related JoVE Video
Development and evaluation of a calibrator material for nucleic acid-based assays for diagnosing aspergillosis.
J. Clin. Microbiol.
PUBLISHED: 04-24-2013
Show Abstract
Hide Abstract
Twelve laboratories evaluated candidate material for an Aspergillus DNA calibrator. The DNA material was quantified using limiting-dilution analysis; the mean concentration was determined to be 1.73 × 10(10) units/ml. The calibrator can be used to standardize aspergillosis diagnostic assays which detect and/or quantify nucleic acid.
Related JoVE Video
Environmental fungicides and triazole resistance in Aspergillus.
Pest Manag. Sci.
PUBLISHED: 04-10-2013
Show Abstract
Hide Abstract
Fungal diseases are problematic in both human health and agriculture. Treatment options are limited and resistance may emerge. The relatively recent recognition of triazole resistance in Aspergillus fumigatus has prompted questioning of the origin of resistance. While multiple mechanisms are described in clinical isolates from triazole-treated patients, some de novo resistance is also recognised, especially attributable to TR34 /L98H. Such strains probably arose in the environment, and, indeed, multiple studies have now demonstrated TR34 /L98H triazole resistance strains of A. fumigatus from soil. Docking and other in vitro studies are consistent with environmental resistance induction through exposure to certain triazole fungicides, notably difenoconazole, propiconazole, epoxiconazole, bromuconazole and tebuconazole. This article addresses the potential implications of this issue for both human health and food security. © 2013 Society of Chemical Industry.
Related JoVE Video
The cdr1B efflux transporter is associated with non-cyp51a-mediated itraconazole resistance in Aspergillus fumigatus.
J. Antimicrob. Chemother.
PUBLISHED: 04-10-2013
Show Abstract
Hide Abstract
Recent increases in triazole resistance in Aspergillus fumigatus have been attributed primarily to target site (cyp51A) mutations. A recent survey of resistant isolates in Manchester showed that >50% of resistant isolates had no mutation in cyp51A or its promoter. We investigated the mechanisms of resistance in clinical azole-resistant isolates without cyp51A mutations.
Related JoVE Video
Intravenous antibiotics reduce the presence of Aspergillus in adult cystic fibrosis sputum.
Thorax
PUBLISHED: 03-19-2013
Show Abstract
Hide Abstract
Pseudomonas aeruginosa and Aspergillus fumigatus frequently co-colonise the airways of patients with cystic fibrosis (CF). This study aimed to assess the impact of short-term administration of intravenous antipseudomonal antibiotics during CF exacerbations on the presence of Aspergillus.
Related JoVE Video
Novel immunologic classification of aspergillosis in adult cystic fibrosis.
J. Allergy Clin. Immunol.
PUBLISHED: 03-10-2013
Show Abstract
Hide Abstract
Patients with cystic fibrosis (CF) demonstrate a wide range of hypersensitivity responses to Aspergillus, beyond allergic bronchopulmonary aspergillosis, which require classification.
Related JoVE Video
Volume dependency for culture of fungi from respiratory secretions and increased sensitivity of Aspergillus quantitative PCR.
Mycoses
PUBLISHED: 03-06-2013
Show Abstract
Hide Abstract
Diagnosis of aspergillosis is often difficult. We compared fungal yields from respiratory specimens using the Health Protection Agency standard culture method (BSOP57), a higher volume undiluted culture method Mycology Reference Centre Manchester (MRCM) and Aspergillus quantitative real time polymerase chain reaction (qPCR). Sputum, bronchial aspirate and bronchoalveolar lavage (BAL) samples (total 23) were collected from aspergillosis patients. One fraction of all samples was cultured using the MRCM method, one BSOP57 and one was used for qPCR. The recovery rate for fungi was significantly higher by MRCM (87%) than by BSOP57 (8.7%) from all 23 specimens. Sputum samples were 44% positive by MRCM compared to no fungi isolated (0%) by BSOP57. Bronchial aspirates were 75% positive by MRCM and 0% by BSOP57. BAL samples were positive in 20% by MRCM and 10% by BSOP57. qPCR was always more sensitive than culture (95.6%) from all samples. In general, over 100 mould colonies (81 Aspergillus fumigatus) were grown using the MRCM method compared with only one colony from BSOP57. This study provides a reference point for standardisation of respiratory sample processing in diagnostic laboratories. Culture from higher volume undiluted respiratory specimens has a much higher yield for Aspergillus than BSOP57. qPCR is much more sensitive than culture and the current UK method requires revision.
Related JoVE Video
Frequency, diagnosis and management of fungal respiratory infections.
Curr Opin Pulm Med
PUBLISHED: 02-16-2013
Show Abstract
Hide Abstract
This review highlights key recent advances in fungal respiratory infections, encompassing developments in epidemiology, diagnostics and management, focussing on Aspergillus, Pneumocystis and Cryptococcus as key pathogens.
Related JoVE Video
Allergic bronchopulmonary aspergillosis and related allergic syndromes.
Semin Respir Crit Care Med
PUBLISHED: 12-13-2011
Show Abstract
Hide Abstract
While allergic bronchopulmonary aspergillosis (ABPA) is well recognized as a fungal complication of asthma, severe asthma with fungal sensitization (SAFS) is not. In ABPA the total immunoglobulin E (IgE) is usually >1,000 IU/mL, whereas in SAFS it is <1,000 IU/mL, and either skin prick tests or fungus-specific IgE tests are positive. ABPA may present with any severity of asthma, and occasionally with no asthma or cystic fibrosis, the other common underlying disease. SAFS is a problem in patients with poorly controlled asthma and occasionally presents in the intensive care unit (ICU). Production of mucous plugs and coughing paroxysms is more common in ABPA. Certain underlying genetic defects seem to underpin these remarkable phenotypic differences. From a management perspective both ABPA and SAFS respond to both high doses of corticosteroids and oral antifungal agents, with ?60% response rate in both ABPA and SAFS with itraconazole. In 50% of patients itraconazole boosts inhaled corticosteroid exposure, sometimes leading to cushingoid features. Second-line therapy data are scant, but we have shown that 70 to 80% of patients who tolerate either voriconazole or posaconazole also respond. Other useful therapies include nebulized hypertonic saline to aid expectoration of thick sputum and long-term azithromycin for its anti-inflammatory effect on the airways. Omaluzimab is useful in some patients with SAFS and occasionally in ABPA. Complications of ABPA include bronchiectasis, typically central in distribution, and chronic pulmonary aspergillosis. Most patients with ABPA and SAFS can be stabilized for long periods with inhaled corticosteroids and itraconazole or another antifungal agent. Novel immunotherapies are on the horizon.
Related JoVE Video
Guidelines for the diagnosis and antibiotic treatment of endocarditis in adults: a report of the Working Party of the British Society for Antimicrobial Chemotherapy.
J. Antimicrob. Chemother.
PUBLISHED: 11-14-2011
Show Abstract
Hide Abstract
The BSAC guidelines on treatment of infectious endocarditis (IE) were last published in 2004. The guidelines presented here have been updated and extended to reflect developments in diagnostics, new trial data and the availability of new antibiotics. The aim of these guidelines, which cover both native valve and prosthetic valve endocarditis, is to standardize the initial investigation and treatment of IE. An extensive review of the literature using a number of different search criteria has been carried out and cited publications used to support any changes we have made to the existing guidelines. Publications referring to in vitro or animal models have only been cited if appropriate clinical data are not available. Randomized, controlled trials suitable for the development of evidenced-based guidelines in this area are still lacking and therefore a consensus approach has again been adopted for most recommendations; however, we have attempted to grade the evidence, where possible. The guidelines have also been extended by the inclusion of sections on clinical diagnosis, echocardiography and surgery.
Related JoVE Video
Azole antifungal resistance today: focus on Aspergillus.
Curr Infect Dis Rep
PUBLISHED: 09-21-2011
Show Abstract
Hide Abstract
Oral triazole therapy is well established for the treatment of invasive aspergillosis (IPA), allergic aspergillosis (ABPA), and chronic pulmonary aspergillosis (CPA), and is often long-term. Resistance to triazole azole antifungal drugs in Aspergillus fumigatus is now a major clinical problem in a number of European locations, in China, Canada and the USA with particularly high frequencies from the north-west of the UK, and The Netherlands. A number of centers are reporting the continuing increasing frequency and evolution of resistance mechanisms in A. fumigatus, in both azole-naïve and patients treated with azoles. The increasing rate of resistance is of concern. A number of resistance mechanisms have been found. The biofilm modality of Aspergillus growth may have a number of therapeutic implications for aspergillosis, including antifungal resistance. Microbiological diagnosis of aspergillosis is limited by poor culture yield, leading to uncertainty about the frequency of triazole resistance. Direct resistance testing in culture-negative clinical samples may add additional insights into the prevalence of azole resistance in A. fumigatus.
Related JoVE Video
Interrogation of related clinical pan-azole-resistant Aspergillus fumigatus strains: G138C, Y431C, and G434C single nucleotide polymorphisms in cyp51A, upregulation of cyp51A, and integration and activation of transposon Atf1 in the cyp51A promoter.
Antimicrob. Agents Chemother.
PUBLISHED: 08-29-2011
Show Abstract
Hide Abstract
Multiple Aspergillus fumigatus isolates from a patient with two aspergillomas complicating chronic pulmonary aspergillosis were pan-azole resistant. Microsatellite typing was identical for all isolates despite major phenotypic and some growth rate differences. Three different cyp51A mutations were found (G138C, Y431C, and G434C), of which the first two were demonstrated by heterologous expression in a hypersusceptible Saccharomyces cerevisiae strain to be at least partly responsible for elevated MICs. cyp51A and cyp51B gene duplication was excluded, but increased expression of cyp51A was demonstrated in three isolates selected for additional study (7-to 13-fold increases). In the isolate with the greatest cyp51A expression, an Aft1 transposon was found inserted 370 bp upstream of the start codon of the cyp51A gene, an integration location never previously demonstrated in Aspergillus. Two transcription start sites were identified at 49 and 136 bp upstream of the start codon. The role of the Aft1 transposon, if any, in modulating cyp51A expression remains to be established. Increased mRNA expression of the transporters AfuMDR1 and AfuMDR4 also was demonstrated in some isolates, which could contribute to azole resistance or simply represent a stress response. The diversity of confirmed and possible azole resistance mechanisms demonstrated in a single series of isogenic isolates is remarkable, indicating the ability of A. fumigatus to adapt in the clinical setting.
Related JoVE Video
Cryptic species and azole resistance in the Aspergillus niger complex.
Antimicrob. Agents Chemother.
PUBLISHED: 07-18-2011
Show Abstract
Hide Abstract
Aspergillus niger is a common clinical isolate. Multiple species comprise the Aspergillus section Nigri and are separable using sequence data. The antifungal susceptibility of these cryptic species is not known. We determined the azole MICs of 50 black aspergilli, 45 from clinical specimens, using modified EUCAST (mEUCAST) and Etest methods. Phylogenetic trees were prepared using the internal transcribed spacer, beta-tubulin, and calmodulin sequences to identify strains to species level and the results were compared with those obtained with cyp51A sequences. We attempted to correlate cyp51A mutations with azole resistance. Etest MICs were significantly different from mEUCAST MICs (P < 0.001), with geometric means of 0.77 and 2.79 mg/liter, respectively. Twenty-six of 50 (52%) isolates were itraconazole resistant by mEUCAST (MICs > 8 mg/liter), with limited cross-resistance to other azoles. Using combined beta-tubulin/calmodulin sequences, the 45 clinical isolates grouped into 5 clades, A. awamori (55.6%), A. tubingensis (17.8%), A. niger (13.3%), A. acidus (6.7%), and an unknown group (6.7%), none of which were morphologically distinguishable. Itraconazole resistance was found in 36% of the isolates in the A. awamori group, 90% of the A. tubingensis group, 33% of the A. niger group, 100% of the A. acidus group, and 67% of the unknown group. These data suggest that cyp51A mutations in section Nigri may not play as important a role in azole resistance as in A. fumigatus, although some mutations (G427S, K97T) warrant further study. Numerous cryptic species are found in clinical isolates of the Aspergillus section Nigri and are best reported as "A. niger complex" by clinical laboratories. Itraconazole resistance was common in this data set, but azole cross-resistance was unusual. The mechanism of resistance remains obscure.
Related JoVE Video
Peripheral neuropathy in patients on long-term triazole antifungal therapy.
J. Antimicrob. Chemother.
PUBLISHED: 06-17-2011
Show Abstract
Hide Abstract
Triazole antifungal drugs are the mainstay of treatment for patients with chronic pulmonary aspergillosis and are often used as steroid-sparing agents in patients with allergic aspergillosis. Peripheral neuropathy (PN) is a rare but reported side effect of triazole therapy in the acute management of invasive fungal infections, but its incidence during long-term triazole treatment for chronic aspergillosis is unknown. The goal of this study was to determine the incidence of PN in this context.
Related JoVE Video
Global burden of chronic pulmonary aspergillosis as a sequel to pulmonary tuberculosis.
Bull. World Health Organ.
PUBLISHED: 04-19-2011
Show Abstract
Hide Abstract
To estimate the global burden of chronic pulmonary aspergillosis (CPA) after pulmonary tuberculosis (PTB), specifically in cases with pulmonary cavitation.
Related JoVE Video
Itraconazole associated quadriparesis and edema: a case report.
J Med Case Rep
PUBLISHED: 04-09-2011
Show Abstract
Hide Abstract
Itraconazole is an anti-fungal agent widely used to treat various forms of mycosis. It is particularly useful in allergic bronchopulmonary aspergillosis and severe asthma with fungal sensitization. Side effects are uncommon and usually mild. Mild neuropathy is noted to occur very rarely. We present an unusual and, to the best of our knowledge, as yet unreported case of severe neuropathy and peripheral edema due to itraconazole in the absence of a concomitant risk factor.
Related JoVE Video
High-frequency triazole resistance found In nonculturable Aspergillus fumigatus from lungs of patients with chronic fungal disease.
Clin. Infect. Dis.
PUBLISHED: 04-07-2011
Show Abstract
Hide Abstract
Oral triazole therapy is well established for the treatment of invasive (IPA), allergic (ABPA), and chronic pulmonary (CPA) aspergillosis, and is often long-term. Triazole resistance rates are rising internationally. Microbiological diagnosis of aspergillosis is limited by poor culture yield, leading to uncertainty about the frequency of triazole resistance.
Related JoVE Video
Pulmonary aspergillosis: an alternative diagnosis to lung cancer after positive [18F]FDG positron emission tomography.
Thorax
PUBLISHED: 04-02-2011
Show Abstract
Hide Abstract
[(18)F]Fluorodexyglucose (FDG) positron emission tomography (PET) scans have significantly improved the diagnosis and staging of lung cancer, but false-positive scans are known to occur due to inflammatory and infectious diseases. Recognition of the conditions leading to false-positive scans is important. Single or multiple pulmonary nodules, with or without cavitation, are classical findings in acute and chronic pulmonary aspergillosis. Clinical features of pulmonary aspergillosis are very similar to those of lung cancer. This report highlights pulmonary aspergillosis as an alternative diagnosis to lung cancer in patients with positive [(18)F]FDG PET scans and the need to strive for presurgical histological diagnosis.
Related JoVE Video
Multicenter, prospective clinical evaluation of respiratory samples from subjects at risk for Pneumocystis jirovecii infection by use of a commercial real-time PCR assay.
J. Clin. Microbiol.
PUBLISHED: 03-02-2011
Show Abstract
Hide Abstract
Pneumocystis jirovecii pneumonia (PCP) is a common opportunistic infection. Microscopic diagnosis, including diagnosis using the Merifluor-Pneumocystis direct fluorescent antigen (MP-DFA) test, has limitations. Real-time PCR may assist in diagnosis, but no commercially validated real-time PCR assay has been available to date. MycAssay Pneumocystis is a commercial assay that targets the P. jirovecii mitochondrial large subunit (analytical detection limit, ? 3.5 copies/?l of sample). A multicenter trial recruited 110 subjects: 54 with transplants (40 with lung transplants), 32 with nonmalignant conditions, 13 with leukemia, and 11 with solid tumors; 9 were HIV positive. A total of 110 respiratory samples (92% of which were bronchoalveolar lavage [BAL] specimens) were analyzed by PCR. Performance was characterized relative to investigator-determined clinical diagnosis of PCP (including local diagnostic tests), and PCR results were compared with MP-DFA test results for 83 subjects. Thirteen of 14 subjects with PCP and 9/96 without PCP (including 5 undergoing BAL surveillance after lung transplantation) had positive PCR results; sensitivity, specificity, and positive and negative predictive values (PPV and NPV, respectively) were 93%, 91%, 59%, and 99%, respectively. Fourteen of 83 subjects for whom PCR and MP-DFA test results were available had PCP; PCR sensitivity, specificity, PPV, and NPV were 93%, 90%, 65%, and 98%, respectively, and MP-DFA test sensitivity, specificity, PPV, and NPV were 93%, 100%, 100%, and 98%. Of the 9 PCR-positive subjects without PCP, 1 later developed PCP. The PCR diagnostic assay compares well with clinical diagnosis using nonmolecular methods. Additional positive results compared with the MP-DFA test may reflect low-level infection or colonization.
Related JoVE Video
Aspergillus fumigatus Bronchopneumonia in a Hellenic Shepherd Dog.
J Am Anim Hosp Assoc
PUBLISHED: 02-10-2011
Show Abstract
Hide Abstract
A 3 yr old intact female Hellenic shepherd dog was referred due to depression, partial anorexia, fever, and a mild productive cough of 2 mo duration. Thoracic radiographs showed increased opacity of all of the left lung lobes. Upon bronchoscopy, a sanguineous, purulent discharge was detected in the tracheal lumen with hyperplastic tissue narrowing the left main stem bronchus. Cultures were positive for bacteria (Bacillus spp. and Clostridium spp.) but negative for fungi. Due to the severity of the lesions, a complete left lung pneumonectomy was performed. Histopathological examination of the excised lung tissues revealed a severe granulomatous bronchopneumonia with numerous alveolar macrophages laden with structures stained positively by periodic acid-Schiff and Grocott stain that had morphology consistent with fungi. PCR and sequencing of internal transcribed spacer regions 1 and 2 from genetic material extracted from paraffin-embedded pulmonary tissue confirmed the presence of Aspergillus fumigatus. Itraconazole was administrated for 5.5 mo and the dog was clinically normal 26 mo after surgery.
Related JoVE Video
Too many mouldy joints - marijuana and chronic pulmonary aspergillosis.
Mediterr J Hematol Infect Dis
PUBLISHED: 01-14-2011
Show Abstract
Hide Abstract
Chronic pulmonary aspergillosis is a progressive debilitating disease with multiple underlying pulmonary diseases described. Here we report the association of chronic pulmonary aspergillosis and long term marijuana smoking in 2 patients and review the literature related to invasive and allergic aspergillosis.
Related JoVE Video
Homogenisation of cystic fibrosis sputum by sonication--an essential step for Aspergillus PCR.
J. Microbiol. Methods
PUBLISHED: 01-14-2011
Show Abstract
Hide Abstract
The importance of Aspergillus as a lung pathogen in cystic fibrosis (CF) is becoming increasingly recognised. However, fungal culture of CF sputum is unreliable and there is no consensus for identifying phenotypes beyond ABPA that may benefit from antifungal therapy. There are no published studies using real-time PCR to detect Aspergillus in CF sputum. The major barrier to sensitive detection of Aspergillus using PCR is sputum homogenisation. This study aimed to optimise sputum homogenisation utilising sonication to improve Aspergillus DNA extraction. Sonication amplitude and duration that enabled sputum homogenisation but ensured preservation of DNA integrity were first determined. 160 sputum samples were collected from CF patients. 49 of the sputum samples were split, one half was used for standard culture and the other half was homogenised with NALC-NaOH before undergoing DNA extraction. The subsequent 111 samples were homogenised with dithiothreitol plus sonication prior to culture and DNA extraction. Real-time PCR targeting a portion of the 18S rDNA of Aspergillus was performed on all DNA extractions. In the 49 samples with no sonication 8 (16%) were culture positive but only 4 of these were PCR positive. However, PCR was positive in 11 culture negative samples. PCR after sonication showed a significant improvement in sensitivity: 33 (30%) were culture and PCR positive, 48 (43%) were culture negative, but PCR positive (p<0.0001) and 30 (27%) were culture and PCR negative. The combination of dithiothreitol and sonication to homogenise sputum increases PCR yield, with PCR being substantially more sensitive than culture.
Related JoVE Video
Efficacy and safety of posaconazole for chronic pulmonary aspergillosis.
Clin. Infect. Dis.
PUBLISHED: 11-05-2010
Show Abstract
Hide Abstract
Chronic pulmonary aspergillosis (CPA) is a severe, progressive respiratory infection characterized by multiple pulmonary cavities and increased levels of antibodies to Aspergillus species. We report the first use of posaconazole in patients with CPA.
Related JoVE Video
Candida tropicalis in human disease.
Crit. Rev. Microbiol.
PUBLISHED: 10-02-2010
Show Abstract
Hide Abstract
Candida tropicalis is one of the more common Candida causing human disease in tropical countries; the frequency of invasive disease varies by geography causing 3--66% of candidaemia. C. tropicalis is taxonomically close to C. albicans and shares many pathogenic traits. C. tropicalis is particularly virulent in neutropenic hosts commonly with hematogenous seeding to peripheral organs. For candidaemia and invasive candidiasis amphotericin B or an echinocandin are recommended as first-line treatment, with extended-spectrum triazoles acceptable alternatives. Primary fluconazole resistance is uncommon but may be induced on exposure. Physicians in regions where C. tropicalis is common need to be mindful of this lesser-described pathogen.
Related JoVE Video
Azole antifungal resistance in Aspergillus fumigatus: 2008 and 2009.
J. Antimicrob. Chemother.
PUBLISHED: 08-20-2010
Show Abstract
Hide Abstract
Resistance to azole antifungal drugs in Aspergillus fumigatus is now a major clinical problem in some locations. Here we update our previous experience with data from 2008-09.
Related JoVE Video
Aspergillus fumigatus allergen expression is coordinately regulated in response to hydrogen peroxide and cyclic AMP.
Clin Mol Allergy
PUBLISHED: 06-30-2010
Show Abstract
Hide Abstract
A. fumigatus has been associated with a wide spectrum of allergic disorders such as ABPA or SAFS. It is poorly understood what allergens in particular are being expressed during fungal invasion and which are responsible for stimulation of immune responses. Study of the dynamics of allergen production by fungi may lead to insights into how allergens are presented to the immune system.
Related JoVE Video
A public resource for metabolic pathway mapping of Aspergillus fumigatus Af293.
Med. Mycol.
PUBLISHED: 05-28-2010
Show Abstract
Hide Abstract
Our understanding of the human pathogenic fungus Aspergillus fumigatus has recently benefitted from much work at the genomics level, including genome sequencing and comparative genome analyses. The next stage in this process is to develop a higher-level appreciation of the organisms biology and to this end the Pathway Tools software has been used to map the metabolic pathways of A. fumigatus Af293. The resulting web-based resource shows 242 pathways which can be viewed at a variety of resolutions. Some pathways have been manually curated (e.g., ergosterol biosynthesis, 4-hydroxymandelate degradation, fatty acid ?-oxidation, fatty acid ?-oxidation, the glyoxylate cycle, palmitate biosynthesis, pyridoxal 5-phosphate salvage, sphingolipid metabolism, ubiquinone biosynthesis and very long chain fatty acid biosynthesis) while others can be used as a starting point for more detailed research. The pathways can be viewed via the Scientific Information:Genomes section of the Aspergillus website ( www.aspergillus.org.uk ).
Related JoVE Video
Molecular detection and identification of zygomycetes species from paraffin-embedded tissues in a murine model of disseminated zygomycosis: a collaborative European Society of Clinical Microbiology and Infectious Diseases (ESCMID) Fungal Infection Study G
J. Clin. Microbiol.
PUBLISHED: 04-07-2010
Show Abstract
Hide Abstract
The present study was performed to assess the interlaboratory reproducibility of the molecular detection and identification of species of Zygomycetes from formalin-fixed paraffin-embedded kidney and brain tissues obtained from experimentally infected mice. Animals were infected with one of five species (Rhizopus oryzae, Rhizopus microsporus, Lichtheimia corymbifera, Rhizomucor pusillus, and Mucor circinelloides). Samples with 1, 10, or 30 slide cuts of the tissues were prepared from each paraffin block, the sample identities were blinded for analysis, and the samples were mailed to each of seven laboratories for the assessment of sensitivity. A protocol describing the extraction method and the PCR amplification procedure was provided. The internal transcribed spacer 1 (ITS1) region was amplified by PCR with the fungal universal primers ITS1 and ITS2 and sequenced. As negative results were obtained for 93% of the tissue specimens infected by M. circinelloides, the data for this species were excluded from the analysis. Positive PCR results were obtained for 93% (52/56), 89% (50/56), and 27% (15/56) of the samples with 30, 10, and 1 slide cuts, respectively. There were minor differences, depending on the organ tissue, fungal species, and laboratory. Correct species identification was possible for 100% (30 cuts), 98% (10 cuts), and 93% (1 cut) of the cases. With the protocol used in the present study, the interlaboratory reproducibility of ITS sequencing for the identification of major Zygomycetes species from formalin-fixed paraffin-embedded tissues can reach 100%, when enough material is available.
Related JoVE Video
Tremor: a newly described adverse event with long-term itraconazole therapy.
J. Neurol. Neurosurg. Psychiatr.
PUBLISHED: 02-27-2010
Show Abstract
Hide Abstract
Itraconazole is a widely prescribed triazole antifungal drug, often given for long periods. The authors report five cases of tremor related to itraconazole therapy, which occurred within 1-12 months of initiating treatment and resolved gradually following itraconazole withdrawal.
Related JoVE Video
Therapy for fungal diseases: opportunities and priorities.
Trends Microbiol.
PUBLISHED: 02-01-2010
Show Abstract
Hide Abstract
This article provides a perspective on the current status of drug therapy for invasive fungal diseases, together with priorities for the future development of novel compounds. Key opportunities for new drugs include production of orally bioavailable agents for the treatment of invasive aspergillosis, invasive candidiasis, cryptococcal meningitis and mucosal and urinary Candida infections. Orally bioavailable agents for the treatment of chronic pulmonary and allergic aspergillosis are also required, as well as new potent drugs against a range of medically important moulds. Antifungal resistance is a problem in certain contexts, but is generally less of a problem than bacterial infections. Earlier and more complete mycological diagnosis and improvements in underlying risk estimation will improve outcomes. The limitations of the current antifungal agents and opportunities for new developments are discussed.
Related JoVE Video
Aspergillus DNA contamination in blood collection tubes.
Diagn. Microbiol. Infect. Dis.
PUBLISHED: 01-07-2010
Show Abstract
Hide Abstract
Fungal polymerase chain reaction (PCR)-based diagnostic methods are at risk for contamination. Sample collection containers were investigated for fungal DNA contamination using real-time PCR assays. Up to 18% of blood collection tubes were contaminated with fungal DNA, probably Aspergillus fumigatus. Lower proportions of contamination in other vessels were observed.
Related JoVE Video
The effects of antifungal therapy on severe asthma with fungal sensitization and allergic bronchopulmonary aspergillosis.
Respirology
PUBLISHED: 11-14-2009
Show Abstract
Hide Abstract
Very little is known about the response rates to or appropriateness of treatment for patients with allergic fungal diseases of the lung. This study assessed the effect of antifungal therapy in patients with severe asthma with fungal sensitization (SAFS) and allergic bronchopulmonary aspergillosis (ABPA).
Related JoVE Video
Incidental finding of colorectal cancer in screening colonoscopy and its cost effectiveness.
Am Surg
PUBLISHED: 09-04-2009
Show Abstract
Hide Abstract
The objective of this study is to measure the risk of colorectal cancer and adenoma with screening colonoscopy and its cost effectiveness. We reviewed the procedure and pathology results of approximately 11,000 asymptomatic patients age 50 to 90 that underwent screening colonoscopy. Among those 11,808 screening colonoscopies performed, advance neoplasm (adenocarcinoma) was detected in 272 (2.3%) patients; age 50 to 90, with mean age of 64-years-old. Fourteen per cent had hyperplastic polyps, 15 per cent had tubular adenoma, and 8.6 per cent villous adenoma. Adenoma with high grade dysplasia was found in 6.6 per cent, and 5.5 per cent had nonadenomatous lesions. Sixty-five of 272 (24%) neoplasms were found proximally. Forty-five of 207 distal neoplasms were found through sigmoidoscopy, nine of 45 (20%) had proximal involvement. Rate of complication during colonoscopy was 0.06 per cent and no patients died. All patients underwent complete colonoscopy, 99.8 per cent were men. Rate of adenocarcinoma from 2000 to 2006 was (24/470, 29/520, 33/891, 37/961, 46/2889, 49/2977, and 54/3100). Screening colonoscopy can detect advanced colorectal neoplasm in asymptomatic adults. The more screening colonoscopy was preformed the earlier the neoplasm was discovered and with better prognosis. Twenty per cent of the patients with distal neoplasms found on sigmoidoscopy had proximal lesions when complete colonoscopies were performed. These findings warrant refinement of the screening recommendations for colorectal cancer.
Related JoVE Video
Toxicodynamics of itraconazole: implications for therapeutic drug monitoring.
Clin. Infect. Dis.
PUBLISHED: 08-18-2009
Show Abstract
Hide Abstract
We explored concentration-toxicity relationships for itraconazole among 216 patients. Logistic regression revealed a progressive increase in the probability of toxicity with increasing concentrations of itraconazole. Classification and regression tree analysis suggested that 17.1 mg/L of itraconazole (measured using a bioassay) was the concentration level at which the population of patients was separated into 2 groups, each with a high and a low probability of toxicity.
Related JoVE Video
A study on Aspergillus species in houses of asthmatic patients from Sari City, Iran and a brief review of the health effects of exposure to indoor Aspergillus.
Environ Monit Assess
PUBLISHED: 07-27-2009
Show Abstract
Hide Abstract
To study the distribution of Aspergillus spp. in outdoor and indoor air of asthmatic patients houses, as well as a review on the health effects of exposure to indoor Aspergillus. Open plates containing malt extract agar media were used to isolate fungi from the indoor (n = 360) and outdoor (n = 180) air of 90 asthmatic patients houses living in Sari City, Iran. Plates were incubated at room temperature for 7-14 days. Cultured Aspergillus spp. were identified by standard mycological techniques. All culture plates grew fungi, a testament to the ubiquitous nature of fungal exposure. Cladosporium spp. (29.2%), Aspergillus spp. (19.0%), and Penicillium spp. (18.3%) were most common inside the houses while Cladosporium spp. (44.5%), Aspergillus spp. (12.4%), and Alternaria spp. (11.1%) were most common outside the houses. Aspergillus flavus (30.1%) and A. fumigatus (23.1%) are the most commonly isolated species in indoor air. Aspergillus flavus (44.5%) and A. fumigatus (42.6%) were the most prevalent Aspergillus spp. outside. The most colony numbers of Aspergillus were isolated from kitchens (30.4%) and the least from bedrooms (21.1%). Aspergillus flavus was the most prevalent species in all sampled rooms except in the kitchen where A. fumigatus was the most common. Aspergillus flavus is the most prevalent species among the Aspergillus spp. in the indoor and outdoor of a warm climate area. In these areas, A. flavus can be a major source of allergen in the air. Therefore, minimizing indoor fungal exposure could play an important role in reducing allergic symptoms in susceptible persons.
Related JoVE Video
Frequency and evolution of Azole resistance in Aspergillus fumigatus associated with treatment failure.
Emerging Infect. Dis.
PUBLISHED: 07-24-2009
Show Abstract
Hide Abstract
Azoles are the mainstay of oral therapy for aspergillosis. Azole resistance in Aspergillus has been reported infrequently. The first resistant isolate was detected in 1999 in Manchester, UK. In a clinical collection of 519 A. fumigatus isolates, the frequency of itraconazole resistance was 5%, a significant increase since 2004 (p<0.001). Of the 34 itraconazole-resistant isolates we studied, 65% (22) were cross-resistant to voriconazole and 74% (25) were cross-resistant to posaconazole. Thirteen of 14 evaluable patients in our study had prior azole exposure; 8 infections failed therapy (progressed), and 5 failed to improve (remained stable). Eighteen amino acid alterations were found in the target enzyme, Cyp51A, 4 of which were novel. A population genetic analysis of microsatellites showed the existence of resistant mutants that evolved from originally susceptible strains, different cyp51A mutations in the same strain, and microalterations in microsatellite repeat number. Azole resistance in A. fumigatus is an emerging problem and may develop during azole therapy.
Related JoVE Video
Fungal rhinosinusitis: a categorization and definitional schema addressing current controversies.
Laryngoscope
PUBLISHED: 06-23-2009
Show Abstract
Hide Abstract
Fungal (rhino-) sinusitis encompasses a wide spectrum of immune and pathological responses, including invasive, chronic, granulomatous, and allergic disease. However, consensus on terminology, pathogenesis, and optimal management is lacking. The International Society for Human and Animal Mycology convened a working group to attempt consensus on terminology and disease classification.
Related JoVE Video
Activity of aminocandin (IP960; HMR3270) compared with amphotericin B, itraconazole, caspofungin and micafungin in neutropenic murine models of disseminated infection caused by itraconazole-susceptible and -resistant strains of Aspergillus fumigatus.
Int. J. Antimicrob. Agents
PUBLISHED: 06-17-2009
Show Abstract
Hide Abstract
Aminocandin (IP960; HMR3270; NXL201) is a new echinocandin with broad-spectrum in vitro activity against Aspergillus and Candida spp. We compared the activity of aminocandin with that of amphotericin B (AmB), itraconazole (ITC) and caspofungin (CAS) in murine models of disseminated aspergillosis against three strains of A. fumigatus, two of which were fully susceptible (AF293 and A1163) and one was resistant to ITC (AF91). Mice were rendered temporarily neutropenic or persistently neutropenic with cyclophosphamide and were infected intravenously 3 days later. Temporarily neutropenic mice were treated with either intraperitoneal (i.p.) AmB (5mg/kg/dose), oral (p.o.) ITC (25mg/kg/dose), intravenous (i.v.) aminocandin (0.25-10mg/kg/dose), i.p. aminocandin (1mg/kg/dose) or solvent control for 9 days. Mice were euthanised 11 days post infection and the kidneys and liver were removed for quantitative culture. Following infection with AF293, only aminocandin 5mg/kg i.v. yielded 100% survival. Aminocandin 1mg/kg i.v., AmB 5mg/kg i.p. or ITC 25mg/kg p.o. were equivalent (P>0.05). Aminocandin 5mg/kg was superior to aminocandin 0.25mg/kg (P<0.0001) as well as all controls (P<0.0001) in reducing mortality. Following infection with AF91, only aminocandin at 5mg/kg and 1mg/kg i.v. yielded 100% survival, which was superior to ITC, aminocandin 0.25mg/kg and controls (all P<0.0001). In the persistently neutropenic model with A1163, aminocandin, CAS and micafungin (2-10mg/kg) were all effective at prolonging survival, with some impact on reducing culture burdens, even with alternate-day dosing (4mg/kg). The only fungicidal regimen was aminocandin 5mg/kg, which sterilised 40% and 50% of mice following infection with AF293 and AF91, respectively. Aminocandin at doses of > or =1mg/kg is highly effective in reducing mortality and organ burden in disseminated infection caused by ITC-susceptible and -resistant A. fumigatus.
Related JoVE Video
Pharmacokinetics and pharmacodynamics of a novel triazole, isavuconazole: mathematical modeling, importance of tissue concentrations, and impact of immune status on antifungal effect.
Antimicrob. Agents Chemother.
PUBLISHED: 05-18-2009
Show Abstract
Hide Abstract
Isavuconazole is a triazole with broad-spectrum activity against medically important fungal pathogens. We investigated the pharmacokinetics and pharmacodynamics of isavuconazole in a murine model of disseminated candidiasis. We determined the pharmacokinetics in both plasma and kidney. The relationship between tissue concentrations and the resultant antifungal effect was described using a mathematical model. The pharmacodynamic parameter that optimally links drug exposure with the antifungal effect was determined using dose fractionation studies. The impact of the immune status of mice receiving isavuconazole was determined in persistently and temporarily neutropenic animals. The pharmacokinetics of 1.6 to 28 mg isavuconazole/kg of body weight were linear. Exposure-response relationships demonstrated near-maximal effect following the administration of >15 mg/kg. The mathematical model showed that exposures in the kidney were 5.77 times higher than those in plasma, and there was persistence of the drug at this site despite concentrations in plasma falling to undetectable levels. The in vitro and in vivo postantifungal effects were 2 to 5 and 8.41 h, respectively. The area under the concentration-time curve (AUC)/MIC ratio was the parameter that optimally linked drug exposure with the observed antifungal effect. The total drug AUC/MIC ratios associated with a 90% probability of survival in temporarily and persistently neutropenic mice were 270 and 670, respectively. Once corrected for protein binding, these values are similar to the magnitude of drug exposure associated with a high probability of a successful therapeutic outcome for other triazoles. This study provides the experimental foundation for the use of isavuconazole in patients with disseminated candidiasis.
Related JoVE Video
Azole-resistance in Aspergillus: proposed nomenclature and breakpoints.
Drug Resist. Updat.
PUBLISHED: 04-18-2009
Show Abstract
Hide Abstract
Reports of itraconazole resistance in Aspergillus fumigatus have been more frequent since the millennium. Identifying azole resistance is critically method dependent; nevertheless reproducible methods, reflective of in vivo outcome, are now in routine use. Some isolates also have elevated MICs to posaconazole and voriconazole. Multiple mechanisms of resistance are now known to be responsible, with differing degrees of azole cross-resistance, including mutations in the Cyp51A gene at G54, L98+TR, G138, M220, G448. Establishing breakpoints for Aspergillus is probably impossible with clinical data alone for multiple reasons yet there is an urgent need to do so. We propose the following breakpoints for A. fumigatus complex using the proposed EUCAST susceptibility testing methodology: for itraconazole and voriconazole, <2 mg/L (susceptible), 2 mg/L (intermediate) and >2 mg/L (resistant); for posaconazole, <0.25, 0.5 and >0.5 mg/L respectively. We recognize that additional work will be needed to confirm these proposed breakpoints, including in vivo and clinical correlative responses. We also propose nomenclature for genotypic resistance, in the event an isolate is not cultured, typified by ITZgR, VCZgI, POSgR (G54W) indicating that the isolate has a G54W substitution with a corresponding phenotype of resistance to itraconazole and posaconazole and intermediate susceptibility to voriconazole.
Related JoVE Video
Using aCGH to study intraspecific genetic variability in two pathogenic molds, Aspergillus fumigatus and Aspergillus flavus.
Med. Mycol.
PUBLISHED: 03-17-2009
Show Abstract
Hide Abstract
We have examined the feasibility of using array comparative genomic hybridization (aCGH) to explore intraspecific genetic variability at the genomic level in two pathogenic molds, Aspergillus fumigatus and Aspergillus flavus. Our analysis showed that strain-specific genes may comprise up to 2% of their genomes in comparison to isolates from different vegetative (heterokaryon) compatibility groups (VCGs). In contrast, isolates with the same VCG affiliations have almost identical gene content. Most isolate-specific genes are annotated as hypothetical and located in a few large subtelomeric indels. The list includes highly polymorphic loci that contain putative het (heterokaryon compatibility) loci, which determine the individuals VCG during parasexual crossing. Incidentally, VCGs in both species seem to be significantly associated with either alpha or HMG mating type (Chi-square test, P=0.05). In conclusion CGH can be used to effectively to identify isolate-specific genes in Aspergillus species. Preliminary evidence suggests that gene flow in both species is largely constrained by VCG boundaries, although further VCG sampling is required to confirm this observation.
Related JoVE Video
Genetic variability of innate immunity impacts human susceptibility to fungal diseases.
Int. J. Infect. Dis.
PUBLISHED: 03-16-2009
Show Abstract
Hide Abstract
Fungi are a major threat in immunocompromised patients. Despite presenting similar degrees of immunosuppression, not all individuals at-risk ultimately develop fungal diseases. The traditional view of immune suppression as a key risk factor for susceptibility to fungal infections needs to be accommodated within new conceptual advances on host immunity and its relationship to fungal disease. The critical role of the immune system emphasizes the contribution of host genetic polymorphisms to fungal disease susceptibility. This review highlights the present knowledge on innate immunity genetics that associates with susceptibility to fungal diseases.
Related JoVE Video
Aspergillus genomes and the Aspergillus cloud.
Nucleic Acids Res.
PUBLISHED: 03-04-2009
Show Abstract
Hide Abstract
Aspergillus Genomes is a public resource for viewing annotated genes predicted by various Aspergillus sequencing projects. It has arisen from the union of two significant resources: the Aspergillus/Aspergillosis website and the Central Aspergillus Data REpository (CADRE). The former has primarily served the medical community, providing information about Aspergillus and associated diseases to medics, patients and scientists; the latter has focused on the fungal genomic community, providing a central repository for sequences and annotation extracted from Aspergillus Genomes. By merging these databases, genomes benefit from extensive cross-linking with medical information to create a unique resource, spanning genomics and clinical aspects of the genus. Aspergillus Genomes is accessible from http://www.aspergillus-genomes.org.uk.
Related JoVE Video
Randomized controlled trial of oral antifungal treatment for severe asthma with fungal sensitization: The Fungal Asthma Sensitization Trial (FAST) study.
Am. J. Respir. Crit. Care Med.
PUBLISHED: 02-04-2009
Show Abstract
Hide Abstract
Some patients with severe asthma are immunologically sensitized to one or more fungi, a clinical entity categorized as severe asthma with fungal sensitization (SAFS). It is not known whether SAFS responds to antifungal therapy.
Related JoVE Video
Aflatoxin and ochratoxin production by Aspergillus species under ex vivo conditions.
Mycopathologia
PUBLISHED: 01-26-2009
Show Abstract
Hide Abstract
Aspergillus species are increasingly important human pathogens. It is not known whether toxic metabolites of many of these pathogenic species can act as virulence factors in aspergillosis. We examined isolates of aflatoxin and ochratoxin-producing species for toxin production in ex vivo conditions. Seven of the 21 aflatoxin-producing isolates screened produced aflatoxin at 35 and 37 degrees C on the general medium yeast extract sucrose agar (YES). However, none of them produced toxin at these temperatures on brain heart infusion agar (BHA), a medium that mimics human tissue, or on BHA with modified pH or sugar levels. Six of the 12 ochratoxin-producing isolates examined produced toxin at 35 degrees C on YES. All three isolates of A. alliaceus produced ochratoxin on BHA or modified BHA at 37 degrees C. One strain of A. pseudoelegans produced a minute amount of ochratoxin on modified BHA at 37 degrees C. These data indicate that aflatoxin is an unlikely virulence, factor but that ochratoxin may be a potential virulence factor in aspergillosis.
Related JoVE Video
Invasive pulmonary aspergillosis 10 years post bone marrow transplantation: a case report.
J Med Case Rep
PUBLISHED: 01-26-2009
Show Abstract
Hide Abstract
Invasive pulmonary aspergillosis is a leading cause of mortality and morbidity in bone marrow transplant recipients. Establishing the diagnosis remains a challenge for clinicians working in acute care setting. However, prompt diagnosis and treatment can lead to favourable outcomes
Related JoVE Video
Hidden killers: human fungal infections.
Sci Transl Med
Show Abstract
Hide Abstract
Although fungal infections contribute substantially to human morbidity and mortality, the impact of these diseases on human health is not widely appreciated. Moreover, despite the urgent need for efficient diagnostic tests and safe and effective new drugs and vaccines, research into the pathophysiology of human fungal infections lags behind that of diseases caused by other pathogens. In this Review, we highlight the importance of fungi as human pathogens and discuss the challenges we face in combating the devastating invasive infections caused by these microorganisms, in particular in immunocompromised individuals.
Related JoVE Video
Sequencing of mitochondrial genomes of nine Aspergillus and Penicillium species identifies mobile introns and accessory genes as main sources of genome size variability.
BMC Genomics
Show Abstract
Hide Abstract
The genera Aspergillus and Penicillium include some of the most beneficial as well as the most harmful fungal species such as the penicillin-producer Penicillium chrysogenum and the human pathogen Aspergillus fumigatus, respectively. Their mitochondrial genomic sequences may hold vital clues into the mechanisms of their evolution, population genetics, and biology, yet only a handful of these genomes have been fully sequenced and annotated.
Related JoVE Video
Aspergillus bronchitis without significant immunocompromise.
Ann. N. Y. Acad. Sci.
Show Abstract
Hide Abstract
Aspergillus bronchitis is poorly understood and described. We extracted clinical data from more than 400 referred patients with persistent chest symptoms who did not fulfill criteria for allergic, chronic, or invasive aspergillosis. Symptomatic patients with a positive culture or real-time PCR for Aspergillus spp. were reviewed. Seventeen patients fulfilled the selected criteria. Fourteen were women, with a mean age of 57 years (range 39-76). Sixteen of the patients had productive cough, eight had voluminous tenacious sputum, and seven had recurrent chest infections. Eight patients had Medical Research Council dyspnea scores of 4-5; 12 had bronchiectasis; and 13 patients grew A. fumigatus, 3 A. niger, and 1 A. terreus. Twelve of the 17 patients (71%) had elevated Aspergillus IgG (47-137 mg/L, mean 89.2) and 5 (29%) had elevated Aspergillus precipitins. Six of 12 (50%) had a major response to antifungal therapy and five of 12 (42%) patients relapsed, requiring long-term therapy. Aspergillus bronchitis is a discrete clinical entity in patients with structural lung disease but who are not significantly immunocompromised. It is distinct from asymptomatic fungal colonization and other forms of aspergillosis, and may respond to antifungal therapy.
Related JoVE Video
Multifocal pulmonary aspergillomas: case series and review.
Ann. N. Y. Acad. Sci.
Show Abstract
Hide Abstract
Multifocal lung parenchymal cavities containing multiple aspergillomas are not well-recognized features of chronic pulmonary aspergillosis (CPA). We identified five patients with multiple cavities containing fungal balls from our accumulated cohort of ?300 patients with CPA. Corticosteroid exposure and radiological and serological characteristics were assessed. The patients, aged 19-55 years, developed 3-11 cavities (or nodules in one case) with thin walls, usually within the lung parenchyma, with very limited pleural involvement. Four had asthma (severe in three) and one had cystic fibrosis; three had allergic bronchopulmonary aspergillosis. All patients had received corticosteroids orally or by inhalation. Four patients had elevated Aspergillus IgG antibodies; one had elevated Aspergillus-specific IgE. Three patients developed azole resistance on antifungal therapy, after benefit, one of whom underwent a successful bilateral lung transplant, later complicated by a fatal mycotic cerebral aneurysm. Multiple aspergillomas is a new distinct manifestation of CPA. The lack of inflammatory response and the distribution of the cavities in the lungs are remarkable. Aspergillus nodules could evolve into cavities with aspergillomas. The management and development of azole resistance in these patients is problematic.
Related JoVE Video
Global burden of allergic bronchopulmonary aspergillosis with asthma and its complication chronic pulmonary aspergillosis in adults.
Med. Mycol.
Show Abstract
Hide Abstract
Allergic bronchopulmonary aspergillosis (ABPA) complicates asthma and may lead to chronic pulmonary aspergillosis (CPA) yet global burdens of each have never been estimated. Antifungal therapy has a place in the management of ABPA and is the cornerstone of treatment in CPA, reducing morbidity and probably mortality. We used the country-specific prevalence of asthma from the Global Initiative for Asthma (GINA) report applied to population estimates to calculate adult asthma cases. From five referral cohorts (China, Ireland, New Zealand, Saudi Arabia and South Africa), we estimated the prevalence of ABPA in adults with asthma at 2.5% (range 0.72-3.5%) (scoping review). From ABPA case series, pulmonary cavitation occurred in 10% (range 7-20%), allowing an estimate of CPA prevalence worldwide using a deterministic scenario-based model. Of 193 million adults with active asthma worldwide, we estimate that 4,837,000 patients (range 1,354,000-6,772,000) develop ABPA. By WHO region, the ABPA burden estimates are: Europe, 1,062,000; Americas, 1,461,000; Eastern Mediterranean, 351,000; Africa, 389,900; Western Pacific, 823,200; South East Asia, 720,400. We calculate a global case burden of CPA complicating ABPA of 411,100 (range 206,300-589,400) at a 10% rate with a 15% annual attrition. The global burden of ABPA potentially exceeds 4.8 million people and of CPA complicating ABPA ˜ 400,000, which is more common than previously appreciated. Both conditions respond to antifungal therapy justifying improved case detection. Prospective population and clinical cohort studies are warranted to more precisely ascertain the frequency of ABPA and CPA in different locations and ethnic groups and validate the model inputs.
Related JoVE Video
Global burden of chronic pulmonary aspergillosis complicating sarcoidosis.
Eur. Respir. J.
Show Abstract
Hide Abstract
Chronic pulmonary aspergillosis (CPA) may complicate pulmonary sarcoidosis. We re-estimated the global burden of sarcoidosis and the burden of CPA complicating sarcoidosis. We searched the literature and reference lists of retrieved papers to identify all published sarcoidosis incidence and prevalence data. We estimated the frequency of CPA from 11 papers relating to >3,000 patients with sarcoidosis to derive CPA patient numbers. We applied an annual attrition rate of 15% (range 10-25%) to estimate the global burden of CPA. We estimate that the annual incidence of sarcoidosis is 344,000 patients worldwide and the prevalence is ?1,238,000 cases, distributed as follows: 165,979 in Europe, 224,000 in the Americas, 492,892 in Africa, 80,023 in the Eastern Mediterranean, 41,660 in the West Pacific and 234,010 in Southeast Asia. CPA complicates sarcoidosis in 3-12% of cases. Using a 6% frequency, we estimate a global burden of 71,907 (range 35,954-143,815 (3-12%)) CPA cases complicating sarcoidosis, with 24% and 37% of cases estimated to be present in the Americas and Africa, because of the higher incidence of sarcoidosis in black people. As CPA responds to long-term antifungal therapy, which may prevent life-threatening haemoptysis, screening periodically for CPA in those with pulmonary sarcoidosis may be important, especially in patients requiring corticosteroid therapy.
Related JoVE Video
Identification of novel genes conferring altered azole susceptibility in Aspergillus fumigatus.
FEMS Microbiol. Lett.
Show Abstract
Hide Abstract
Azoles are currently the mainstay of antifungal treatment both in agricultural and in clinical settings. Although the target site of azole action is well studied, the basis of azole resistance and the ultimate mode of action of the drug in fungi are poorly understood. To gain a deeper insight into these aspects of azole action, restriction-mediated plasmid integration (REMI) was used to create azole sensitive and resistant strains of the clinically important fungus Aspergillus fumigatus. Four azole sensitive insertions and four azole-resistant insertions were characterized. Three phenotypes could be re-created in wild-type AF210 by reintegration of rescued plasmid and a further four could be confirmed by complementation of the mutant phenotype with a copy of the wild-type gene predicted to be disrupted by the original insertional event. Six insertions were in genes not previously associated with azole sensitivity or resistance. Two insertions occur in transporter genes that may affect drug efflux, whereas others may affect transcriptional regulation of sterol biosynthesis genes and NADH metabolism in the mitochondrion. Two insertions are in genes of unknown function.
Related JoVE Video
The performance of real-time PCR, galactomannan, and fungal culture in the diagnosis of invasive aspergillosis in ventilated patients with chronic obstructive pulmonary disease (COPD).
Mycopathologia
Show Abstract
Hide Abstract
Emerging reports have associated chronic pulmonary obstructive disease (COPD) with invasive aspergillosis (IA), particularly in patients treated with mechanical ventilation and/or corticosteroids. This is a multicentre study in which COPD patients demonstrating a new lung infiltrate while being mechanically ventilated were prospectively evaluated for the presence of IA. From the 47 patients studied, Aspergillus fumigatus was recovered in culture in two patients (4.2%). While serum galactomannan (GM) was negative for 94% of patients, GM levels in respiratory samples were >0.5, >1.0 and >1.5 for 74.5, 40.5, and 21.3% of patients, respectively. PCR was positive for 10 patients in the study but did not differentiate Aspergillus colonization from infection. The combination of PCR and GM in respiratory samples may be an interesting alternative to diagnose IA in COPD patients.
Related JoVE Video

What is Visualize?

JoVE Visualize is a tool created to match the last 5 years of PubMed publications to methods in JoVE's video library.

How does it work?

We use abstracts found on PubMed and match them to JoVE videos to create a list of 10 to 30 related methods videos.

Video X seems to be unrelated to Abstract Y...

In developing our video relationships, we compare around 5 million PubMed articles to our library of over 4,500 methods videos. In some cases the language used in the PubMed abstracts makes matching that content to a JoVE video difficult. In other cases, there happens not to be any content in our video library that is relevant to the topic of a given abstract. In these cases, our algorithms are trying their best to display videos with relevant content, which can sometimes result in matched videos with only a slight relation.