%0 Journal Article %~ PubMed %A Chen, Sharon C-A %A Slavin, Monica A %A Heath, Christopher H %A Playford, E Geoffrey %A Byth, Karen %A Marriott, Deborah %A Kidd, Sarah E %A Bak, Narin %A Currie, Bart %A Hajkowicz, Krispin %A Korman, Tony M %A McBride, William Jh %A Meyer, Wieland %A Murray, Ronan %A Sorrell, Tania C %A , on behalf of the Australia and New Zealand Mycoses Interest Group (ANZMIG)-Cryptococcus Study %T Clinical manifestations of Cryptococcus gattii infection: determinants of neurological sequelae and death. %B Clinical Infectious Diseases %D 2012 %C United States %I Oxford University Press %V 55 %N 6 %P 789-798 %@ 1537-6591 %X Background.???Longer-term morbidity and outcomes of Cryptococcus gattii (CG) infection are not described. We analyzed clinical, microbiological and outcome data in Australian patients followed for 12 months to identify prognostic determinants.Methods.???Culture-confirmed CG cases from 2000-2007 were retrospectively evaluated. Clinical, microbiological, radiological and outcome data were recorded at diagnosis and at 6 weeks, 6 months and 12 months. Clinical and laboratory variables associated with mortality and with death and/or neurological sequelae were determined.Results.???Annual CG infection incidence was 0.61/10(6) population. Sixty-two of 86 (72%) patients had no immunocompromise; 6 of 24 immunocompromised hosts had idiopathic CD4 lymphopenia and 1, HIV/AIDS. Clinical and microbiological characteristics of infection were similar in immunocompromised and healthy hosts. Isolated lung, combined lung and central nervous system (CNS), and CNS only, disease was reported in 12%, 51% and 34%, respectively. Complications in CNS disease included raised intracranial pressure (42%), hydrocephalus (30%), neurological deficits (27%; 6% developed during therapy) and immune reconstitution-like syndrome (11%). Geometric mean serum cryptococcal antigen (CRAG) titres in CNS disease were 563.9 (vs. 149.3 in isolated lung infection). Patient immunocompromise was associated with increased mortality risk. Initial cerebrospinal fluid CRAG titre of ???256 predicted death and/or neurological sequelae (P = .05).Conclusions.???Neurological CG disease predominates in Australian endemic setting. Lumbar puncture and cerebral imaging, especially if serum CRAG titres are ???512, are essential. Long-term follow up is required to detect late neurological complications. Immune system evaluation is important since host immunocompromise is associated with reduced survival. %Z FOR Codes: 104 %0 Journal Article %~ PubMed %A McMullan, Brendan J %A Halliday, Catriona %A Sorrell, Tania C %A Judd, David %A Sleiman, Sue %A Marriott, Debbie %A Olma, Tom %A Chen, Sharon C-A %T Clinical utility of the cryptococcal antigen lateral flow assay in a diagnostic mycology laboratory. %B PLoS One %D 2012 %C United States %I Public Library of Science %V 7 %N 11 %P e49541 %@ 1932-6203 %X %Z FOR Codes: 1108 %0 Journal Article %~ PubMed %A Aminnejad, Mojgan %A Diaz, Mara %A Arabatzis, Michael %A Castaņeda, Elizabeth %A Lazera, Marcia %A Velegraki, Aristea %A Marriott, Deborah %A Sorrell, Tania C %A Meyer, Wieland %T Identification of Novel Hybrids Between Cryptococcus neoformans var. grubii VNI and Cryptococcus gattii VGII. %B Mycopathologia %D 2012 %C Netherlands %I Springer Netherlands %V 173 %N 5-6 %P 337-346 %@ 1573-0832 %X Cryptococcus neoformans and Cryptococcus gattii are pathogenic yeasts causing meningoencephalitis in immunocompromised and immunocompetent hosts. The fungus is typically haploid, and sexual reproduction occurs normally between individuals with opposite mating types, ?? and a. C. neoformans var. grubii (serotype A) is comprised of molecular types VNI, VNII, and VNB, and C. neoformans var. neoformans (serotype D) contains the molecular type VNIV. Additionally, diploid or aneuploid AD hybrids (VNIII) have been reported. C. gattii contains the molecular types VGI, VGII, VGIII, and VGIV, which encompass both serotypes B and C. To identify possible hybrid strains, URA5-RFLP analysis was performed on 350 globally obtained clinical, environmental, and veterinary isolates. Four clinical isolates from cerebrospinal fluid showed combination patterns of C. neoformans var. grubii and C. gattii: Brazil (n??=??2), Colombia (n??=??1), and India (n??=??1). These strains were monokaryotic and diploid or aneuploid. M13 PCR fingerprinting showed that they contained fragments of both proposed parental groups. Luminex IGS genotyping identified these isolates as hybrids with two different molecular type combinations: three VNI/VGII and one VNI/VGI. Blue color development on CGB agar was delayed in three isolates and absent in one. C. gattii-specific PCR confirmed the presence of C. gattii in the hybrids. CAP59 allele-specific PCR revealed that all the hybrids contained both serotype A and B alleles. Determination of mating-type allelic patterns by PCR revealed that the isolates were ??A aB. This is the first study discovering novel natural hybrids between C. neoformans molecular type VNI and C. gattii molecular type VGII. %Z FOR Codes: 110899 %0 Journal Article %~ PubMed %A Marais, Ben %A Crawford, John %A Iredell, Jon %A Ward, Michael %A Simpson, Steve %A Gilbert, Lyn %A Griffiths, Paul %A Kamradt-Scott, Adam %A Colagiuri, Ruth %A Jones, Cheryl %A Sorrell, Tania %T One world, one health: beyond the Millennium Development Goals. %B Lancet %D 2012 %C United Kingdom %I The Lancet Publishing Group %V 380 %N 9844 %P 805-806 %@ 0140-6736 %X %Z FOR Codes: 60502 60299 110899 %0 Journal Article %~ PubMed %A Wang, He %A Xiao, Meng %A Kong, Fanrong %A Chen, Sharon %A Dou, Hong-Tao %A Sorrell, Tania %A Li, Ruo-Yu %A Xu, Ying-Chun %T Accurate and practical identification of 20 Fusarium species by seven-locus sequence analysis and reverse line blot hybridization, and an in vitro antifungal susceptibility study. %B Journal of Clinical Microbiology %D 2011 %C United States %I American Society for Microbiology %V 49 %N 5 %P 1890-1898 %@ 0095-1137 %X Eleven reference and 25 clinical isolates of Fusarium were subject to multilocus DNA sequence analysis to determine the species and haplotypes of the fusarial isolates from Beijing and Shandong, China. Seven loci were analyzed: the translation elongation factor 1 alpha gene (EF-1?); the nuclear rRNA internal transcribed spacer (ITS), large subunit (LSU), and intergenic spacer (IGS) regions; the second largest subunit of the RNA polymerase gene (RPB2); the calmodulin gene (CAM); and the mitochondrial small subunit (mtSSU) rRNA gene. We also evaluated an IGS-targeted PCR/reverse line blot (RLB) assay for species/haplotype identification of Fusarium. Twenty Fusarium species and seven species complexes were identified. Of 25 clinical isolates (10 species), the Gibberella (Fusarium) fujikuroi species complex was the commonest (40%) and was followed by the Fusarium solani species complex (FSSC) (36%) and the F. incarnatum-F. equiseti species complex (12%). Six FSSC isolates were identified to the species level as FSSC-3+4, and three as FSSC-5. Twenty-nine IGS, 27 EF-1?, 26 RPB2, 24 CAM, 18 ITS, 19 LSU, and 18 mtSSU haplotypes were identified; 29 were unique, and haplotypes for 24 clinical strains were novel. By parsimony informative character analysis, the IGS locus was the most phylogenetically informative, and the rRNA gene regions were the least. Results by RLB were concordant with multilocus sequence analysis for all isolates. Amphotericin B was the most active drug against all species. Voriconazole MICs were high (>8 ?g/ml) for 15 (42%) isolates, including FSSC. Analysis of larger numbers of isolates is required to determine the clinical utility of the seven-locus sequence analysis and RLB assay in species classification of fusaria. %Z FOR Codes: 110899 %0 Journal Article %~ PubMed %A Ngamskulrungroj, Popchai %A Price, Jennifer %A Sorrell, Tania %A Perfect, John R %A Meyer, Wieland %T Cryptococcus gattii virulence composite: candidate genes revealed by microarray analysis of high and less virulent Vancouver island outbreak strains. %B PLoS One %D 2011 %C United States %I Public Library of Science %V 6 %N 1 %P e16076 %@ 1932-6203 %X Human and animal cryptococcosis due to an unusual molecular type of Cryptococcus gattii (VGII) emerged recently on Vancouver Island, Canada. Unlike C. neoformans, C. gattii causes disease mainly in immunocompetent hosts, despite producing a similar suite of virulence determinants. To investigate a potential relationship between the regulation of expression of a virulence gene composite and virulence, we took advantage of two subtypes of VGII (a and b), one highly virulent (R265) and one less virulent (R272), that were identified from the Vancouver outbreak. By expression microarray analysis, 202 genes showed at least a 2-fold difference in expression with 108 being up- and 94 being down-regulated in strain R265 compared with strain R272. Specifically, expression levels of genes encoding putative virulence factors (e.g. LAC1, LAC2, CAS3 and MPK1) and genes encoding proteins involved in cell wall assembly, carbohydrate and lipid metabolism were increased in strain R265, whereas genes involved in the regulation of mitosis and ergosterol biosynthesis were suppressed. In vitro phenotypic studies and transcription analysis confirmed the microarray results. Gene disruption of LAC1 and MPK1 revealed defects in melanin synthesis and cell wall integrity, respectively, where CAS3 was not essential for capsule production. Moreover, MPK1 also controls melanin and capsule production and causes a severe attenuation of the virulence in a murine inhalational model. Overall, this study provides the basis for further genetic studies to characterize the differences in the virulence composite of strains with minor evolutionary divergences in gene expression in the primary pathogen C. gattii, that have led to a major invasive fungal infection outbreak. %Z FOR Codes: 110899 %0 Journal Article %~ PubMed %A Morrissey, C Orla %A Chen, Sharon C-A %A Sorrell, Tania C %A Bradstock, Kenneth F %A Szer, Jeffrey %A Halliday, Catriona L %A Gilroy, Nicole M %A Schwarer, Anthony P %A Slavin, Monica A %T Design issues in a randomized controlled trial of a pre-emptive versus empiric antifungal strategy for invasive aspergillosis in patients with high-risk hematologic malignancies. %B Leukemia & Lymphoma %D 2011 %C Switzerland %I Informa Healthcare %V 52 %N 2 %P 179-193 %@ 1029-2403 %X Invasive aspergillosis (IA) is a major cause of mortality in patients with hematological malignancies, due largely to the inability of traditional culture and biopsy methods to make an early or accurate diagnosis. Diagnostic accuracy studies suggest that Aspergillus galactomannan (GM) enzyme immunoassay (ELISA) and Aspergillus PCR-based methods may overcome these limitations, but their impact on patient outcomes should be evaluated in a diagnostic randomized controlled trial (D-RCT). This article describes the methodology of a D-RCT which compares a new pre-emptive strategy (GM-ELISA- and Aspergillus PCR-driven antifungal therapy) with the standard fever-driven empiric antifungal treatment strategy. Issues including primary end-point and patient selection, duration of screening, choice of tests for the pre-emptive strategy, antifungal prophylaxis and bias control, which were considered in the design of the trial, are discussed. We suggest that the template presented herein is considered by researchers when evaluating the utility of new diagnostic tests (ClinicalTrials.gov number, NCT00163722). %Z FOR Codes: 110202 111201 %0 Journal Article %~ PubMed %A Konstantinos, Anastasios %A Simpson, Graham %A Sorrell, Tania %A Marais, Ben J %T Doing the right thing for tuberculosis control in the Torres Strait Islands. %B Medical Journal of Australia %D 2011 %C Australia %I Australasian Medical Publishing Company Pty. Ltd. %V 195 %N 9 %P 512 %@ 1326-5377 %X %Z FOR Codes: 111701 110203 110309 %0 Journal Article %~ PubMed %A Chen, Sharon C-A %A Slavin, Monica A %A Sorrell, Tania C %T Echinocandin antifungal drugs in fungal infections: a comparison. %B Drugs %D 2011 %C New Zealand %I Adis International Ltd. %V 71 %N 1 %P 11-41 %@ 0012-6667 %X This review compares the pharmacology, spectrum of antifungal activity, pharmacokinetic and pharmacodynamic properties, safety and clinical efficacy of the three licensed echinocandins: caspofungin, micafungin and anidulafungin. Echinocandins inhibit the synthesis of 1,3-?-D-glucan, an essential component of the fungal cell wall, and represent a valuable treatment option for fungal infections. The echinocandins exhibit potent in vitro and in vivo fungicidal activity against Candida species, including azole-resistant pathogens. For all agents, strains with drug minimum inhibitory concentrations (MICs) of ? 2??g/mL are considered susceptible; the MIC at which 90% of isolates tested were inhibited (MIC??) values are typically <2??g/mL but 100-fold higher MIC?? values are seen with Candida parapsilosis (1-2??g/mL) and Candida guilliermondii (1-4??g/mL). Activity is comparable between the three agents, although limited data indicate that anidulafungin may have low MICs against C. parapsilosis and Candida glabrata strains that demonstrate elevated MICs to caspofungin and micafungin. All three drugs have good fungistatic activity against Aspergillus spp., although minimal effective concentrations of micafungin and anidulfungin are 2- to 10-fold lower than those for caspofungin. Synergistic/additive in vitro effects of echinocandins when combined with a polyene or azole have been observed. Clinical resistance to the echinocandins is rare despite case reports of caspofungin resistance in several Candida spp. Resistance has been attributed to mutations in the FKS1 gene within two hot spot regions, leading to amino acid substitutions, mostly at position 645 (serine), yet not all FKS1 mutants have caspofungin MICs of >2??g/mL. Of the three echinocandins, the in vitro ''paradoxical effect'' (increased growth at supra-MIC drug concentrations) is observed least often with anidulafungin. All echinocandins have low oral bioavailability, and distribute well into tissues, but poorly into the CNS and eye. Anidulafungin is unique in that it undergoes elimination by chemical degradation in bile rather than via hepatic metabolism, has a lower maximum concentration and smaller steady state under the concentration-time curve but longer half-life than caspofungin or micafungin. In children, dosing should be based on body surface area. Daily doses of caspofungin (but not micafungin and anidulafungin) should be decreased (from 50 to 35?mg) in moderate liver insufficiency. All echinocandins display concentration-dependent fungicidal (for Candida) or fungistatic (for Aspergillus) activity. The postantifungal effect is 0.9-20 hours against Candida and <0.5 hours against Aspergillus. The echinocandins are well tolerated with few serious drug-drug interactions since they are not appreciable substrates, inhibitors or inducers of the cytochrome P450 or P-glycoprotein systems. In parallel with the greater clinical experience with caspofungin, this agent has a slightly higher potential for adverse effects/drug-drug interactions, with the least potential observed for anidulafungin. Caspofungin (but not micafungin or anidulafungin) dosing should be increased if coadministered with rifampicin and there are modest interactions of caspofungin with calcineurin inhibitors. All three agents are approved for the treatment of oesophageal candidiasis, candidaemia and other select forms of invasive candidiasis. Only micafungin is licensed for antifungal prophylaxis in stem cell transplantation, whereas caspofungin is approved for empirical therapy of febrile neutropenia. Caspofungin has been evaluated in the salvage and primary therapy of invasive aspergillosis. Combination regimens incorporating an echinocandin showing promise in the treatment of aspergillosis. However, echinocandins remain expensive to use. %Z FOR Codes: 60505 111501 %0 Journal Article %~ PubMed %A Ferguson, P E %A Gilroy, N M %A Sloots, T P %A Nissen, M D %A Dwyer, D E %A Sorrell, T C %T Evaluation of a clinical scoring system and directed laboratory testing for respiratory virus infection in hematopoietic stem cell transplant recipients. %B Transplant Infectious Disease %D 2011 %C United States %I Wiley-Blackwell Publishing, Inc. %V 13 %N 5 %P 448-455 %@ 1399-3062 %X A simple clinical screening (CS) tool for respiratory virus (RV) infection was introduced and evaluated in a single hematology ward, as part of a strategy to reduce nosocomial RV infection. Up to 6 clinical symptoms or signs were scored and a predefined threshold score of ??? 2 prompted paired nose/throat swab (NTS) collection for RV testing. The criterion standard for RV infection was positive immunofluorescence (IF) or polymerase chain reaction (PCR) for 7 and 15 viruses, respectively. The tool was shown to be most beneficial at excluding infection at a threshold score of 1 (negative predictive value [NPV] 89%, [95% confidence interval 78-96%], sensitivity 85% [70-94%], specificity 35% [27-43%]), compared with a score of 2 (NPV 85% [76-91%], sensitivity 63% [46-77%], specificity 57% [48-65%]) at a prevalence of 22%. The tool''s ability to diagnose infection was limited (positive predictive value 27% and 29% at thresholds 1 and 2). The sensitivity of IF compared with PCR was 45% for the 7 viruses common to both, and 23% for the extended virus panel detected by PCR. An algorithm incorporating CS, paired NTS collection at a threshold of 1 symptom or sign, and sensitive testing including PCR can guide infection control measures in hospitalized hematopoietic stem cell transplant recipients. %Z FOR Codes: 110804 110202 %0 Journal Article %~ PubMed %A Wang, Tao %A Kong, Fanrong %A Chen, Sharon %A Xiao, Meng %A Sorrell, Tania %A Wang, Xiaoyan %A Wang, Shuo %A Sintchenko, Vitali %T Improved identification of Gordonia, Rhodococcus and Tsukamurella species by 5'-end 16S rRNA gene sequencing. %B Pathology %D 2011 %C United Kingdom, Australia %I Informa Healthcare %V 43 %N 1 %P 58-63 %@ 0031-3025 %X The identification of fastidious aerobic Actinomycetes such as Gordonia, Rhodococcus, and Tsukamurella has remained a challenge leading to clinically significant misclassifications. This study is intended to examine the feasibility of partial 5''-end 16S rRNA gene sequencing for the identification of Gordonia, Rhodococcus, and Tsukamurella, and defined potential reference sequences for species from each of these genera. %Z FOR Codes: 1110 602 %0 Journal Article %~ PubMed %A Zuo, Xiaoming %A Djordjevic, Julianne T %A Bijosono Oei, Johanes %A Desmarini, Desmarini %A Schibeci, Stephen D %A Jolliffe, Katrina A %A Sorrell, Tania C %T Miltefosine Induces Apoptosis-like Cell Death in Yeast via Cox9p in Cytochrome c Oxidase. %B Molecular Pharmacology %D 2011 %C United States %I American Society for Pharmacology and Experimental Therapeutics %V 80 %N 3 %P 476-485 %@ 1521-0111 %X Miltefosine has antifungal properties and potential for development as a therapeutic for invasive fungal infections. However, its mode of action in fungi is poorly understood. We demonstrate that miltefosine is rapidly incorporated into yeast, where it penetrates the mitochondrial inner membrane, disrupting mitochondrial membrane potential and leading to an apoptosis-like cell death. COX9, which encodes subunit VIIa of the cytochrome c oxidase (COX) complex in the electron transport chain of the mitochondrial membrane, was identified as a potential target of miltefosine from a genomic library screen of the model yeast Saccharomyces cerevisiae. When overexpressed in S. cerevisiae, COX9, but not COX7 or COX8, led to a miltefosine-resistant phenotype. The effect of miltefosine on COX activity was assessed in cells expressing different levels of COX9. Miltefosine inhibited COX activity in a dose-dependent manner in Cox9p-positive cells. This inhibition most likely contributed to the miltefosine-induced apoptosis-like cell death. %Z FOR Codes: 601 605 %0 Journal Article %~ PubMed %A Outhred, Alexander C %A Watts, Matthew R %A Chen, Sharon C-A %A Sorrell, Tania C %T Nocardia infections of the face and neck. %B Current Infectious Disease Reports %D 2011 %C United States %I Current Medicine Group LLC %V 13 %N 2 %P 132-140 %@ 1534-3146 %X Involvement of the soft tissues of the face and neck by Nocardia spp. is uncommon. We review the epidemiology, clinical features, diagnosis, and management of such infections in the setting of primary cutaneous nocardiosis and disseminated disease. Although immune compromise is an important risk factor for these infections, they also occur in healthy individuals. Infection may arise through direct inoculation following injury or by hematogenous spread from a primary site, usually the lung. The rare variant of lymphocutaneous disease-cervicofacial nocardiosis-typically affects children, but can occur in adults. The diagnosis is made by conventional microscopy and culture, but radiological imaging is useful to delineate disease extent, and molecular methods are increasingly assisting the diagnosis by providing rapid detection and identification of the pathogen. Sulfonamides remain the preferred treatment for many cases and are an important component of the therapeutic armamentarium. Other therapeutic options include minocycline, the carbapenems, and linezolid. %Z FOR Codes: 110899 110309 %0 Journal Article %~ PubMed %A Chen, Sharon C-A %A Blyth, Christopher C %A Sorrell, Tania C %A Slavin, Monica A %T Pneumonia and Lung Infections due to Emerging and Unusual Fungal Pathogens. %B Seminars in Respiratory and Critical Care Medicine %D 2011 %C United States %I Thieme Medical Publishers %V 32 %N 6 %P 703-716 %@ 1098-9048 %X Invasive mold infections affecting the lungs are increasing in incidence and diversity. Severely immunocompromised patients are particularly vulnerable to infection from unusual, normally nonpathogenic fungi that are found naturally in the environment. Certain fungi such as SCEDOSPORIUM and the dematiaceous fungi also cause lung disease in hosts without overt immune compromise. The impacts of these emerging pathogens range from airway colonization to locally invasive lung, and disseminated, disease. Diagnosis requires isolation and identification of the etiologic agent by either or both phenotypic and molecular biology methods. Evidence of tissue invasion on histopathology is often required to distinguish infection from colonization. Diagnostic imaging techniques are nonspecific, and there are no reliable serological biomarkers of infection. Many rare molds and yeasts demonstrate reduced in vitro susceptibility to antifungal agents. Although amphotericin B formulations remain clinically useful for many of these infections, voriconazole and posaconazole are more effective for some of these difficult-to-treat pathogens. Surgical resection of diseased tissue and support of the host immune system are often required to optimize outcomes. %Z FOR Codes: 110899 %0 Journal Article %~ PubMed %A Chayakulkeeree, Methee %A Johnston, Simon Andrew %A Oei, Johanes Bijosono %A Lev, Sophie %A Williamson, Peter Richard %A Wilson, Christabel Frewen %A Zuo, Xiaoming %A Leal, Ana Lusia %A Vainstein, Marilene Henning %A Meyer, Wieland %A Sorrell, Tania Christine %A May, Robin Charles %A Djordjevic, Julianne Teresa %T SEC14 is a specific requirement for secretion of phospholipase B1 and pathogenicity of Cryptococcus neoformans. %B Molecular Microbiology %D 2011 %C United Kingdom %I Wiley-Blackwell Publishing Ltd. %V 80 %N 4 %P 1088-1101 %@ 0950-382X %X Secreted phospholipase B1 (CnPlb1) is essential for dissemination of Cryptococcus neoformans to the central nervous system (CNS) yet essential components of its secretion machinery remain to be elucidated. Using gene deletion analysis we demonstrate that CnPlb1 secretion is dependent on the CnSEC14 product, CnSec14-1p. CnSec14-1p is a homologue of the phosphatidylinositol transfer protein ScSec14p, which is essential for secretion and viability in Saccharomyces cerevisiae. In contrast to CnPlb1, neither laccase 1-induced melanization within the cell wall nor capsule induction were negatively impacted in CnSEC14-1 deletion mutants (Cn??sec14-1 and Cn??sec14-1Cn??sfh5). Similar to the CnPLB1 deletion mutant (Cn??plb1), Cn??sec14-1 was hypovirulent in mice and did not disseminate to the CNS by day 14 post infection. Furthermore, macrophage expulsion of live Cn??sec14-1 and Cn??plb1 (vomocytosis) was reduced. Individual deletion of CnSEC14-2, a closely related CnSEC14-1 homologue, and CnSFH5, a distantly related SEC fourteen like homologue, did not abrogate CnPlb1 secretion or virulence. However, reconstitution of Cn??sec14-1 with CnSEC14-1 or CnSEC14-2 restored both phenotypes, consistent with functional genetic redundancy. We conclude that CnPlb1 secretion is SEC14-dependent and that C. neoformans preferentially exports virulence determinants to the cell periphery via distinct pathways. We also demonstrate that CnPlb1 secretion is essential for vomocytosis. %Z FOR Codes: 110899 60108 %0 Book Section %A Fox, Deborah S %A Djordjevic, Julianne %A Sorrell, Tania %T Signaling cascades and enzymes as Cryptococcus virulence factors %B Cryptococcus: From Human Pathogen to Model Yeast %D 2011 %C United States %I American Society for Microbiology %V %N %P 217-234 %@ 9781555815011 %E Heitman, Joseph %E Kozel, Thomas R. %E Kwon-Chung, Kyung J. %E Perfect, John R. %E Casadevall, Arturo %X %Z FOR Codes: 110804 %0 Journal Article %~ PubMed %A Guo, Li-Na %A Xiao, Meng %A Kong, Fanrong %A Chen, Sharon C-A %A Wang, He %A Sorrell, Tania C %A Jiang, Wei %A Dou, Hong-Tao %A Li, Ruo-Yu %A Xu, Ying-Chun %T Three-locus identification, genotyping, and antifungal susceptibilities of medically important Trichosporon species from China. %B Journal of Clinical Microbiology %D 2011 %C United States %I American Society for Microbiology %V 49 %N 11 %P 3805-3811 %@ 0095-1137 %X Three reference and 45 clinical isolates of Trichosporon were analyzed by conventional phenotypic and molecular methods to determine the species and genotypes of Trichosporon isolates from China. Target loci for molecular methods included the internal transcribed spacer (ITS) region, the D1/D2 domain of the 26S rRNA gene, and the intergenic spacer 1 (IGS1) region. Identification of eight Trichosporon species was achieved, of which Trichosporon asahii was the most common. Of the sequence-based molecular methods, the one targeting the D1/D2 domain assigned 97.9% (47/48) of isolates (seven species) correctly, while tests targeting both the ITS and IGS1 regions correctly identified all 48 isolates. The commercial API 20C AUX and Vitek 2 Compact YST systems correctly identified 91.9% and 73% of isolates when their biochemical profiles were queried against those of species contained in the databases, respectively, and misidentified 63.6% and 36.4% of isolates of species that were unclaimed by the databases, respectively. The predominant genotype among T. asahii clinical isolates, genotype 4 (51.4%), is rarely found in other countries. Voriconazole and itraconazole were the most active drugs in vitro against all the Trichosporon species tested, while caspofungin and amphotericin B demonstrated poor activity. %Z FOR Codes: 110899 %0 Journal Article %~ PubMed %A Chen, Sharon C-A %A Playford, E Geoffrey %A Sorrell, Tania C %T Antifungal therapy in invasive fungal infections. %B Current opinion in pharmacology %D 2010 %C United Kingdom %I Elsevier Ltd. %V 10 %N 5 %P 522-30 %@ 1471-4973 %X Early treatment of invasive fungal infections (IFIs) is essential for optimal clinical outcomes. Standard antifungal drugs (polyenes, azoles and echinocandins) are not predictably effective against emerging yeasts and filamentous fungi and may cause undesirable side effects. Species identification can guide antifungal selection for invasive candidiasis, but not less common moulds such as Scedosporium and Fusarium spp. Management strategies targeted to those at highest risk (prophylaxis), those with clinical signs of infection not responsive to antibacterials (empiric therapy) and those with occult infection (asymptomatic but with positive fungal biomarkers) produce better outcomes than therapy predicated on identification of a fungal pathogen, but require comparative evaluation. Appropriate dosing and consideration of pharmacokinetic parameters (including therapeutic drug monitoring) are important with newer triazoles. New therapies such as addition of the iron chelator, deferasirox, in the treatment of zygomycosis in diabetic patients, appear promising but additional agents with new targets of action are urgently needed. %Z FOR Codes: 1103 1112 %0 Journal Article %~ PubMed %A Slavin, Monica A %A Sorrell, Tania C %A Marriott, Deborah %A Thursky, Karin A %A Nguyen, Quoc %A Ellis, David H %A Morrissey, C Orla %A Chen, Sharon C-A %A , Australian Candidemia Study, Australasian Society for Infectious Diseases %T Candidaemia in adult cancer patients: risks for fluconazole-resistant isolates and death. %B The Journal of Antimicrobial Chemotherapy %D 2010 %C United Kingdom %I Oxford University Press %V 65 %N 5 %P 1042-1051 %@ 1460-2091 %X BACKGROUND: Candidaemia in cancer patients is associated with increasing fluconazole resistance. Models for predicting such isolates and their clinical impact are required. METHODS: Clinical, treatment and outcome data from a population-based candidaemia survey (2001-2004) were collected at 5 and 30 days after diagnosis. Speciation and antifungal susceptibility testing was performed. RESULTS: There were 138 candidaemia episodes (33% Candida albicans) in adults with haematological malignancies and 150 (51% C. albicans) in adults with solid organ malignancies. Thirty-nine isolates had fluconazole MICs of >or=64 mg/L and 40 had MICs of 16-32 mg/L (predominantly Candida glabrata and Candida krusei). By multivariate analysis, triazole therapy, gastrointestinal tract (GIT) surgery in the 30 days before candidaemia and age >65 years were predictive of fluconazole-resistant candidaemia. Thirty day crude mortality was 40% in haematology patients and 45% in oncology patients. Fluconazole-resistant isolates were associated with increased risk of mortality by univariate (P = 0.04) and Kaplan-Meier survival analyses. By Cox proportional hazards modelling, the strongest predictors of mortality at onset of candidaemia were invasive ventilation, elevated creatinine, intensive care unit (ICU) admission and receipt of systemic triazoles or corticosteroids in the previous 30 days. Removal of a central venous access device (CVAD) at or within 5 days of onset was associated with decreased mortality. CONCLUSIONS: Risk factors for fluconazole-resistant candidaemia in adults with cancer include fluconazole/triazole exposure and GIT surgery. ICU admission, invasive ventilation, renal impairment, age >65 years and prior exposure to corticosteroids and triazoles are risk factors for death. CVAD removal reduced mortality. These findings should be integrated into surveillance and treatment algorithms. %Z FOR Codes: 111706 111299 %0 Journal Article %~ PubMed %A Blyth, Christopher C %A Middleton, Peter G %A Harun, Azian %A Sorrell, Tania C %A Meyer, Wieland %A Chen, Sharon C-A %T Clinical associations and prevalence of Scedosporium spp. in Australian cystic fibrosis patients: identification of novel risk factors? %B Medical Mycology %D 2010 %C United Kingdom %I Informa Healthcare %V 48 %N 1 %P S37-S44 %@ 1460-2709 %X Risk factors for the association of Scedosporium in cases of cystic fibrosis (CF) and its clinical implications are poorly understood. Clinical, lung function and laboratory data of adult CF patients in Sydney (April 2008-March 2009) were prospectively analysed for such risk factors. Expectorated sputa were cultured for bacteria and examined for fungi using standard mycological and Scedosporium-selective media, and by an internal transcribed spacer region-targeted multiplex PCR assay. Scedosporium spp. (n = 4 each of Scedosporium prolificans, Scedosporium aurantiacum and Pseudallescheria boydii/ Scedosporium apiospermum complex [non-S. aurantiacum]) were recovered from 12 of 69 (17.4%) patients. Samples of 11 of the patients yielded isolates on Scedosporium- selective media (vs. 6 [8.7%] by non-selective culture) and one additional patient was noted by PCR. Of these patients, 83.3% were co-colonized with other moulds, most frequently Aspergillus fumigatus. Colonization was not associated with best FEV(1)/predicted, corticosteroid or antifungal therapies. By univariate analysis, patients with Scedosporium colonization were significantly less likely to be colonized with mucoid Pseudomonas aeruginosa (P = 0.025), while prior therapy with antistaphylococcal penicillins was a risk factor for colonization (P = 0.045). Bacterial colonization and antimicrobial exposure likely influence Scedosporium colonization, which is optimally detected with selective media. Studies are required to confirm independent risk factors for Scedosporium colonization and to determine its impact on lung disease. %Z FOR Codes: 110309 110899 %0 Journal Article %~ PubMed %A Perfect, John R %A Dismukes, William E %A Dromer, Francoise %A Goldman, David L %A Graybill, John R %A Hamill, Richard J %A Harrison, Thomas S %A Larsen, Robert A %A Lortholary, Olivier %A Nguyen, Minh-Hong %A Pappas, Peter G %A Powderly, William G %A Singh, Nina %A Sobel, Jack D %A Sorrell, Tania C %T Clinical practice guidelines for the management of cryptococcal disease: 2010 update by the Infectious Diseases Society of America. %B Clinical Infectious Diseases %D 2010 %C United States %I Oxford University Press %V 50 %N 3 %P 291-322 %@ 1537-6591 %X Cryptococcosis is a global invasive mycosis associated with significant morbidity and mortality. These guidelines for its management have been built on the previous Infectious Diseases Society of America guidelines from 2000 and include new sections. There is a discussion of the management of cryptococcal meningoencephalitis in 3 risk groups: (1) human immunodeficiency virus (HIV)-infected individuals, (2) organ transplant recipients, and (3) non-HIV-infected and nontransplant hosts. There are specific recommendations for other unique risk populations, such as children, pregnant women, persons in resource-limited environments, and those with Cryptococcus gattii infection. Recommendations for management also include other sites of infection, including strategies for pulmonary cryptococcosis. Emphasis has been placed on potential complications in management of cryptococcal infection, including increased intracranial pressure, immune reconstitution inflammatory syndrome (IRIS), drug resistance, and cryptococcomas. Three key management principles have been articulated: (1) induction therapy for meningoencephalitis using fungicidal regimens, such as a polyene and flucytosine, followed by suppressive regimens using fluconazole; (2) importance of early recognition and treatment of increased intracranial pressure and/or IRIS; and (3) the use of lipid formulations of amphotericin B regimens in patients with renal impairment. Cryptococcosis remains a challenging management issue, with little new drug development or recent definitive studies. However, if the diagnosis is made early, if clinicians adhere to the basic principles of these guidelines, and if the underlying disease is controlled, then cryptococcosis can be managed successfully in the vast majority of patients. %Z FOR Codes: 60505 60502 304 %0 Journal Article %~ PubMed %A Lau, Anna %A Halliday, Catriona %A Chen, Sharon C-A %A Playford, E Geoffrey %A Stanley, Keith %A Sorrell, Tania C %T Comparison of Whole Blood, Serum and Plasma for Early Detection of Candidemia by Multiplex-Tandem PCR. %B Journal of clinical microbiology %D 2010 %C United States %I American Society for Microbiology %V 48 %N 3 %P 811-6 %@ 0095-1137 %X We applied multiplex-tandem PCR (MT-PCR) to 255 EDTA whole-blood specimens, 29 serum specimens, and 24 plasma specimens from 109 patients with Candida bloodstream infection (candidemia) to determine whether a diagnosis could be expedited in comparison with the time to diagnosis by the use of standard blood culture. Overall, the MT-PCR performed better than blood culture with DNA extracted from whole blood from 52/74 (70%) patients, accelerating the time to detection (blood culture flagging) and determination of the pathogenic species (by use of the API 32C system [bioM??rieux, Marcy l''Etoile, France]) by up to 4 days (mean, 2.2 days; range, 0.5 to 8 days). Candida DNA was detected more often in serum (71%) and plasma (75%) than in whole blood (54%), although relatively small numbers of serum and plasma specimens were tested. The sensitivity, specificity, positive predictive value, and negative predictive value of the assay with whole blood were 75%, 97%, 95%, and 85%, respectively. Fungal DNA was not detected by MT-PCR in 6/24 (25%) whole-blood samples drawn simultaneously with the positive blood culture sample. MT-PCR performed better with whole-blood specimens stored at -20 degrees C (75%) and when DNA was extracted within 1 week of sampling (66%). The molecular and culture identification results correlated for 61 of 66 patients (92%); one discrepant result was due to misidentification by culture. All but one sample from 53 patients who were at high risk of candidemia but did not have proven disease were negative by MT-PCR. The results demonstrate the good potential of MT-PCR to detect candidemia, to provide Candida species identification prior to blood culture positivity, and to provide improved sensitivity when applied to with serum and plasma specimens. %Z FOR Codes: 1110 %0 Journal Article %~ PubMed %A Blyth, C C %A Harun, A %A Middleton, P G %A Sleiman, S %A Lee, O %A Sorrell, T C %A Meyer, W %A Chen, S C A %T Detection of occult Scedosporium species in respiratory tract specimens from patients with cystic fibrosis (CF) by use of selective media. %B Journal of Clinical Microbiology %D 2010 %C United States %I American Society for Microbiology %V 48 %N 1 %P 314-316 %@ 0095-1137 %X Respiratory samples from cystic fibrosis outpatients were cultured on Sabouraud''s dextrose agar containing antibiotics, Mycosel and Scedosporium-selective media (SceSel+). Thirty-two of 218(14.7%) specimens from 11/69(15.9%) patients yielded a Scedosporium spp., most frequently Scedosporium aurantiacum (17/218). Scedosporium was recovered on SceSel+, Mycosel and SABD from 90.6%, 50.0% and 46.9% specimens, respectively. %Z FOR Codes: 60501 110309 110801 %0 Journal Article %~ PubMed %A Hale, Katherine A %A Shaw, Peter J %A Dalla-Pozza, Luciano %A Macintyre, Chandini Raina %A Isaacs, David %A Sorrell, Tania C %T Epidemiology of paediatric invasive fungal infections and a case-control study of risk factors in acute leukaemia or post stem cell transplant. %B British journal of haematology %D 2010 %C United Kingdom %I Wiley-Blackwell Publishing Ltd. %V 149 %N 2 %P 263-72 %@ 1365-2141 %X Patients aged 0-18 years with confirmed or possible invasive fungal infection were identified by medical record and database searches. Cases with an underlying diagnosis of acute leukaemia or following stem cell transplantation were included in a case control study. Controls included all other children with acute leukaemia or stem cell transplant in the corresponding time period. Variables collected included demographics, underlying disease risk and status, organ impairment, admission to intensive care unit, fungal infection details and certain transplant variables. Risk factors for development of invasive fungal infection were examined using logistic regression. There were 106 cases of invasive fungal infection during the study. The incidence of invasive fungal infection was 21% in acute lymphoblastic leukaemia, 15% in acute myeloid leukaemia and 25% following stem cell transplantation. Sixty per cent were neutropenic at diagnosis and 39% had concomitant bacteremia. High risk acute lymphoblastic leukaemia, relapsed disease, intensive care admission and graft-versus-host disease were significantly associated with development of invasive fungal infection on multivariate analysis. These associations provide new information on paediatric invasive fungal infections and warrant further study; caution should be encouraged when extrapolating from adult studies. %Z FOR Codes: 1114 %0 Journal Article %A Sorrell, Tania %A Djordjevic, Julianne %A Chen, Sharon %A Jolliffe, Katrina A %T Fungal phospholipid metabolism for antifungal drug discovery %B Microbiology Australia %D 2010 %C Australia %I The Australian Society for Microbiology Inc. %V 31 %N %P 93-94 %@ 1324-4272 %X %Z FOR Codes: 60505 %0 Journal Article %~ PubMed %A Xiao, Meng %A Kong, Fanrong %A Sorrell, Tania C %A Cao, Yongyan %A Lee, Ok Cha %A Liu, Ying %A Sintchenko, Vitali %A Chen, Sharon C A %T Identification of Pathogenic Nocardia Species by Reverse Line Blot Hybridization Targeting the 16S rDNA and 16S-23S rDNA Spacer Regions. %B Journal of clinical microbiology %D 2010 %C United States %I American Society for Microbiology %V 48 %N 2 %P 503-11 %@ 0095-1137 %X Although 16S rRNA gene sequence analysis is employed most often for the definitive identification of Nocardia species, alternate molecular methods and polymorphisms in other gene targets have also enabled species determinations. We evaluated a combined Nocardia PCR-based reverse line blot (RLB) hybridization assay based on 16S and 16S-23S rRNA gene spacer region polymorphisms to identify 12 American Type Culture Collection and 123 clinical Nocardia isolates representing 14 species; results were compared with results from 16S rRNA gene sequencing. Thirteen 16S rRNA gene-based (two group-specific and 11 species-specific) and five 16S-23S spacer-targeted (two taxon-specific and three species-specific) probes were utilized. 16S rRNA gene-based probes correctly identified 124 of 135 isolates (sensitivity, 92%) but were unable to identify Nocardia paucivorans strains (n = 10 strains) and a Nocardia asteroides isolate with a novel 16S rRNA gene sequence. Nocardia farcinica and Nocardia cyriacigeorgica strains were identified by the sequential use of an N. farcinica-"negative" probe and a combined N. farcinica/N. cyriacigeorgica probe. The assay specificity was high (99%) except for weak cross-reactivity between the Nocardia brasiliensis probe with the Nocardia thailandica DNA product; however, cross-hybridization with closely related nontarget species may occur. The incorporation of 16S-23S rRNA gene spacer-based probes enabled the identification of all N. paucivorans strains. The overall sensitivity using both probe sets was >99%. Both N. farcinica-specific 16S-23S rRNA gene spacer-directed probes were required to identify all N. farcinica stains by using this probe set. The study demonstrates the utility of a combined PCR/RLB assay for the identification of clinically relevant Nocardia species and its potential for studying subtypes of N. farcinica. Where species assignment is ambiguous or not possible, 16S rRNA gene sequencing is recommended. %Z FOR Codes: 110303 60501 60411 %0 Journal Article %~ PubMed %A Chen, Sharon C-A %A Biswas, Chayanika %A Bartley, Robyn %A Widmer, Fred %A Pantarat, Namfon %A Obando, Daniel %A Djordjevic, Julianne T %A Ellis, David H %A Jolliffe, Katrina A %A Sorrell, Tania C %T In vitro antifungal activities of bis(alkylpyridinium)alkane compounds against pathogenic yeasts and molds. %B Antimicrobial agents and chemotherapy %D 2010 %C United States %I American Society for Microbiology %V 54 %N 8 %P 3233-40 %@ 1098-6596 %X Ten bis(alkylpyridinium)alkane compounds were tested for antifungal activity against 19 species (26 isolates) of yeasts and molds. We then determined the MICs and minimum fungicidal concentrations (MFCs) of four of the most active compounds (compounds 1, 4, 5, and 8) against 80 Candida and 20 cryptococcal isolates, in comparison with the MICs of amphotericin B, fluconazole, itraconazole, voriconazole, posaconazole, and caspofungin, using Clinical Laboratory and Standards Institutes broth microdulition M27-A3 (yeasts) or M38-A2 (filamentous fungi) susceptibility protocols. The compounds were more potent against Candida and Cryptococcus spp. (MIC range, 0.74 to 27.9 microg/ml) than molds (0.74 to 59.7 microg/ml). MICs against Exophiala were 0.37 to 5.9 microg/ml and as low as 1.48 microg/ml for Scedosporium but >or=25 microg/ml for zygomycetes, Aspergillus, and Fusarium spp. Compounds 1, 4, 5, and 8 exhibited good fungicidal activity against Candida and Cryptococcus, except for Candida parapsilosis (MICs of >44 mug/ml). Geometric mean (GM) MICs were similar to those of amphotericin B and lower than or comparable to fluconazole GM MICs but 10- to 100-fold greater than those for the other azoles. GM MICs against Candida glabrata were <1 microg/ml, significantly lower than fluconazole GM MICs (P<0.001) and similar to those of itraconazole, posaconazole, and voriconazole (GM MIC range of 0.4 to 1.23 microg/ml). The GM MIC of compound 4 against Candida guilliermondii was lower than that of fluconazole (1.69 microg/ml versus 7.48 microg/ml; P=0.012). MICs against Cryptococcus neoformans and Cryptococcus gattii were similar to those of fluconazole. The GM MIC of compound 4 was significantly higher for C. neoformans (3.83 mug/ml versus 1.81 microg/ml for C. gattii; P=0.015). This study has identified clinically relevant in vitro antifungal activities of novel bisalkypyridinium alkane compounds. %Z FOR Codes: 30499 605 1108 %0 Journal Article %~ PubMed %A Playford, E G %A Nimmo, G R %A Tilse, M %A Sorrell, T C %T Increasing incidence of candidaemia: long-term epidemiological trends, Queensland, Australia, 1999-2008. %B The Journal of Hospital Infection %D 2010 %C United Kingdom %I WB Saunders Co. Ltd. %V 76 %N 1 %P 46-51 %@ 1532-2939 %X Given variability in the epidemiology of candidaemia and a relative paucity of contemporary longitudinal data, a passive laboratory-based surveillance study was performed to assess the epidemiology of candidaemia in all public healthcare facilities in Queensland, Australia over the period 1999-2008. Demographic and microbiological data on all candidaemia episodes, together with appropriate denominators (admissions and patient-days), were collected from laboratory and administrative information systems. From 1999 to 2008, 1137 episodes occurred (overall incidence-density: 0.45 per 10 000 patient-days) with a 3.5-fold increase in density (P<0.0001 for trend). Candidaemia episodes originating in traditional high-risk areas either decreased (haemato-oncology and paediatric wards) or remained stable (intensive care units). Episodes on adult medical/surgical wards increased significantly over time, accounting for 60% of the total by 2008. The relative proportion caused by Candida albicans decreased and Candida parapsilosis increased (both P<0.01). The proportion of fluconazole-resistant isolates did not change. The increasing occurrence of candidaemia outside traditional high-risk areas and the emergence of C. parapsilosis present new challenges for preventive and early intervention strategies. %Z FOR Codes: 111706 %0 Journal Article %~ PubMed %A Gilbert, Nicole M %A Donlin, Maureen J %A Gerik, Kimberly J %A Specht, Charles A %A Djordjevic, Julianne T %A Wilson, Christabel F %A Sorrell, Tania C %A Lodge, Jennifer K %T KRE genes are required for beta-1,6-glucan synthesis, maintenance of capsule architecture and cell wall protein anchoring in Cryptococcus neoformans. %B Molecular Microbiology %D 2010 %C United Kingdom %I Wiley-Blackwell Publishing Ltd. %V 76 %N 2 %P 517-534 %@ 0950-382X %X The polysaccharide beta-1,6-glucan is a major component of the cell wall of Cryptococcus neoformans, but its function has not been investigated in this fungal pathogen. We have identified and characterized seven genes, belonging to the KRE family, which are putatively involved in beta-1,6-glucan synthesis. The H99 deletion mutants kre5Delta and kre6Deltaskn1Delta contained less cell wall beta-1,6-glucan, grew slowly with an aberrant morphology, were highly sensitive to environmental and chemical stress and were avirulent in a mouse inhalation model of infection. These two mutants displayed alterations in cell wall chitosan and the exopolysaccharide capsule, a primary cryptococcal virulence determinant. The cell wall content of the GPI-anchored phospholipase B1 (Plb1) enzyme, which is required for cryptococcal cell wall integrity and virulence, was reduced in kre5Delta and kre6Deltaskn1Delta. Our results indicate that KRE5, KRE6 and SKN1 are involved in beta-1,6-glucan synthesis, maintenance of cell wall integrity and retention of mannoproteins and known cryptococcal virulence factors in the cell wall of C. neoformans. This study sets the stage for future investigations into the function of this abundant cell wall polymer. %Z FOR Codes: 60503 %0 Journal Article %~ PubMed %A Playford, E Geoffrey %A Lipman, Jeff %A Sorrell, Tania C %T Management of invasive candidiasis in the intensive care unit. %B Drugs %D 2010 %C New Zealand %I Adis International Ltd. %V 70 %N 7 %P 823-839 %@ 0012-6667 %X Invasive candidiasis (IC) is an important infection in the intensive care unit (ICU) setting given its association with poor clinical outcomes. The epidemiology of IC is complex and, although incompletely elucidated, is characterized by considerable regional and temporal variability. Overall, there appears to be an increase in the incidence of IC and a change in distribution of the causative Candida spp. Of particular concern is an increase in the proportion of episodes caused by Candida glabrata, which is associated with reduced susceptibility to azole antifungal agents. The management of IC has been aided by the availability of several new antifungal agents. In particular, given their broad spectrum of activity and low toxicity, the use of echinocandins as first-line therapy is increasing, especially in settings where fluconazole-resistant Candida spp. are prevalent. Fluconazole remains a reliable agent where an azole-susceptible pathogen is confirmed or in settings where resistance is uncommon. Lipid formulations of amphotericin B are now generally reserved as second-line or salvage therapy. Voriconazole and posaconazole currently enjoy limited use for IC in the ICU setting. Although the poor outcomes associated with IC are, in part, related to the severity of underlying host factors, it is clear that optimization of treatment-related factors is also important. In particular, the speed of initiation of antifungal therapy and the achievement of pharmacodynamic parameters both influence outcomes. The most difficult challenge is early initiation of an effective antifungal drug, given the slow turnaround time and lack of sensitivity of conventional culture-based diagnostic techniques. New approaches, such as non-culture-based assays and/or clinical risk-predictive models are required to better target prophylactic, pre-emptive and empirical antifungal strategies. %Z FOR Codes: 111706 110310 %0 Journal Article %~ PubMed %A Krockenberger, Mark B %A Malik, Richard %A Ngamskulrungroj, Popchai %A Trilles, Luciana %A Escandon, Patricia %A Dowd, Susan %A Allen, Chris %A Himmelreich, Uwe %A Canfield, Paul J %A Sorrell, Tania C %A Meyer, Wieland %T Pathogenesis of Pulmonary Cryptococcus gattii Infection: A Rat Model. %B Mycopathologia %D 2010 %C Netherlands %I Springer Netherlands %V 170 %N %P 315-30 %@ 1573-0832 %X A model of pulmonary cryptococcosis in immunocompetent rats was developed to better understand the virulence of Cryptococcus gattii. Six isolates were studied, representing four molecular genotypes (VGI-MAT??, VGIIa-MAT??, VGIIa-MAT a, VGIIb-MAT??), obtained from Australia, Vancouver (Canada) and Colombia. These originated from human patients, a cat and the environment and were administered intratracheally (i.t.) or transthoracically into Fischer 344 or Wistar-Furth rats in doses varying from 10(4) to 10(7) colony-forming units (CFU) in 0.1 ml of saline. With the exception of animals given the VGIIa-MAT a isolate, rats consistently became ill or died of progressive cryptococcal pneumonia following i.t. doses exceeding 10(7) CFU. Affected lungs increased in weight up to tenfold and contained numerous circumscribed, gelatinous lesions. These became larger and more extensive, progressing from limited hilar and/or tracheal lesions, to virtually confluent gelatinous masses. Disease was localized to the lungs for at least 3-4 weeks, with dissemination to the brain occurring in some animals after day 29. The dose-response relationship was steep for two VGI isolates studied (human WM179, environmental WM276); doses up to 10(6) CFU i.t. did not produce lesions, while 10(7) or more yeast cells produced progressive pneumonia. Intratracheal inoculation of rats with C. gattii provides an excellent model of human pulmonary cryptococcosis in healthy hosts, mimicking natural infections. Disease produced by C. gattii in rats is distinct from that caused by C. neoformans in that infections are progressive and ultimately fatal. %Z FOR Codes: 605 1108 %0 Journal Article %~ PubMed %A Playford, Elliott Geoffrey %A Lipman, Jeff %A Sorrell, Tania C %T Prophylaxis, empirical and preemptive treatment of invasive candidiasis. %B Current opinion in critical care %D 2010 %C United States %I Lippincott Williams & Wilkins %V 16 %N 5 %P 470-4 %@ 1531-7072 %X Invasive candidiasis remains an important infection for ICU patients, associated with poor clinical outcomes. It has been increasingly recognized that the traditional paradigm of culture-directed antifungal treatment is unsatisfactory, and that earlier antifungal intervention strategies, such as prophylaxis, preemptive therapy, and empiric therapy, are required to improve patient outcomes. The purpose of this review is to summarize the recent supportive evidence for such strategies and to highlight the current challenges in their implementation. %Z FOR Codes: 1103 1108 %0 Journal Article %~ PubMed %A Sorrell, Tania C %A Chen, Sharon C-A %T Recent advances in management of cryptococcal meningitis: commentary. %B F1000 Medicine Reports %D 2010 %C United Kingdom %I Biology Reports Ltd. %V 2 %N %P 82 %@ 1740-309X %X Cryptococcal meningitis remains a substantial health burden with high morbidity, particularly in developing countries. Antifungal treatment regimens are guided by host factors, severity of illness (including presence of complications), and causative cryptococcal species. Recent clinical studies indicate the need for rapidly fungicidal induction therapy regimens using amphotericin B in combination with flucytosine for optimal outcomes. Maintenance therapy with fluconazole is necessary until recovery of immune function. Cryptococcus gattii meningitis requires prolonged induction/eradication therapy. Prompt control of raised intracranial pressure or hydrocephalus is essential. Clinicians should be vigilant for immune restoration-like features. Adjuvant surgery, corticosteroids, and/or recombinant interferon-gamma may be required for large cryptococcomas, cerebral edema, or refractory infection. %Z FOR Codes: 60505 %0 Journal Article %~ PubMed %A Wang, Xiaoyan %A Xiao, Meng %A Kong, Fanrong %A Sintchenko, Vitali %A Wang, Huiping %A Wang, Bin %A Lian, Shi %A Sorrell, Tania %A Chen, Sharon %T Reverse line blot hybridization and DNA sequencing studies of the 16S-23S rRNA gene intergenic spacer regions of five emerging pathogenic Nocardia species. %B Journal of Medical Microbiology %D 2010 %C United Kingdom %I Society for General Microbiology %V 59 %N Pt 5 %P 548-555 %@ 1473-5644 %X The objective of this study was to examine DNA sequence polymorphisms in the 16S-23S rRNA gene intergenic spacer (ITS) regions of five emerging pathogenic Nocardia species: Nocardia beijingensis, Nocardia blacklockiae, Nocardia thailandica, Nocardia elegans and Nocardia vinacea. A set of six isolates belonging to the species of interest and 135 isolates belonging to other Nocardia species was studied. A PCR-based reverse line blot (RLB) hybridization assay incorporating species- or intraspecies ITS rRNA gene operon-specific probes was then developed for species identification. Substantial intraspecies sequence variation among different ITS operons was identified. Four sequence types of N. thailandica, eight sequence types of N. beijingensis (four types for each of two strains) and five sequence types of N. blacklockiae, N. elegans and N. vinacea were found. The results represent the first evidence of ITS sequence heterogeneity in emerging species of Nocardia. By incorporating species/operon-specific probes into a RLB assay, unique RLB patterns were identified for each of the species and every sequence type. The PCR/RLB assay demonstrated high specificity and showed promise in both the identification and genotyping of Nocardia species. More detailed studies of the polymorphism within the ITS locus may further advance our capacity to reliably identify and subtype medically important Nocardia species. %Z FOR Codes: 60503 %0 Journal Article %~ PubMed %A Kong, Fanrong %A Wang, Huiping %A Zhang, Erqing %A Sintchenko, Vitali %A Xiao, Meng %A Sorrell, Tania C %A Chen, Xiaoyou %A Chen, Sharon C-A %T SecA1 Gene Sequence Polymorphisms for Species Identification of Nocardia and Recognition of Intraspecies Genetic Diversity. %B Journal of clinical microbiology %D 2010 %C United States %I American Society for Microbiology %V 48 %N 11 %P 3928-34 %@ 0095-1137 %X Sequence analysis of the Nocardia essential secretory protein SecA1 gene (secA1) for species identification of 120 American Type Culture Collection (ATCC) and clinical isolates of Nocardia (16 species) was studied in comparison with 5''-end 606-bp 16S rRNA gene sequencing. Species determination by both methods was concordant for all 10 ATCC strains. secA1 gene sequencing provided the same species identification as 16S rRNA gene analysis for 94/110 (85.5%) clinical isolates. However, 40 (42.6%) isolates had sequences with <99.0% similarity to archived secA1 sequences for the species, including 29 Nocardia cyriacigeorgica (96.6 to 98.9% similarity) and 4 Nocardia veterana (91.5 to 98.9% similarity) strains. Discrepant species identification was obtained for 16 (14.5%) clinical isolates, including 13/23 Nocardia nova strains (identified as various Nocardia species by secA1 sequencing) and 1 isolate each of Nocardia abscessus (identified as Nocardia asiatica), Nocardia elegans (Nocardia africana), and Nocardia transvalensis (Nocardia blacklockiae); both secA1 gene sequence analysis and deduced amino acid sequence analysis determined the species to be different from those assigned by 16S rRNA gene sequencing. The secA1 locus showed high sequence diversity (66 sequence or genetic types versus 40 16S rRNA gene sequence types), which was highest for N. nova (14 secA1 sequence types), followed by Nocardia farcinica and N. veterana (n = 7 each); there was only a single sequence type among eight Nocardia paucivorans strains. The secA1 locus has potential for species identification as an adjunct to 16S rRNA gene sequencing but requires additional deduced amino acid sequence analysis. It may be a suitable marker for phylogenetic/subtyping studies. %Z FOR Codes: 110303 110801 110309 %0 Journal Article %~ PubMed %A Blyth, Christopher C %A Gomes, Lavier %A Sorrell, Tania C %A da Cruz, Melville %A Sud, Archana %A Chen, Sharon C-A %T Skull-base osteomyelitis: fungal versus bacterial infection. %B Clinical Microbiology and Infection %D 2010 %C United Kingdom, Swit %I Wiley-Blackwell Publishing Ltd. %V 17 %N 2 %P 306-311 %@ 1469-0691 %X Skull-base osteomyelitis (SBO) occurs secondary to invasive bacterial and fungal infection. Distinguishing between fungal and bacterial aetiologies of SBO has significant therapeutic implications. An 18-year (1990-2007) retrospective review of patients with SBO presenting to Westmead Hospital was performed. Epidemiological, clinical, laboratory and radiology data were collated. Twenty-one patients (median age 58 years) with SBO were identified: ten (48%) had bacterial and 11 (52%) had fungal SBO. Diabetes mellitus (57%) and chronic otitis externa (33%) were the most frequent co-morbidities; immunosuppression was present in five cases (24%). Cranial nerve deficits occurred in ten (48%) patients. The commonest pathogens were Pseudomonas aeruginosa (50% bacterial SBO) and a zygomycete (55% fungal SBO). Compared to bacterial SBO, fungal SBO was more frequently associated with underlying chronic sinusitis, sinonasal pain, facial/periorbital swelling and nasal stuffiness or discharge and the absence of purulent ear discharge (all p <0.05). Bacterial SBO was more frequently associated with deafness, ear pain or ear discharge (all p <0.05). Median time to presentation was longer in patients with bacterial SBO (26.3 weeks vs. 8.1 weeks, p 0.08). Overall 6-month survival was 88% (14/18 patients). All four deaths occurred in patients with fungal SBO. Immunosuppression was a risk factor for death (p <0.05). Early diagnostic sampling is recommended in patients at increased risk of fungal SBO to enable optimal antimicrobial and surgical management. %Z FOR Codes: 605 %0 Journal Article %~ PubMed %A Bialasiewicz, Seweryn %A Rockett, Rebecca %A Whiley, David W %A Abed, Yacine %A Allander, Tobias %A Binks, Michael %A Boivin, Guy %A Cheng, Allen C %A Chung, Ju-Young %A Ferguson, Patricia E %A Gilroy, Nicole M %A Leach, Amanda J %A Lindau, Cecilia %A Rossen, John W %A Sorrell, Tania C %A Nissen, Michael D %A Sloots, Theo P %T Whole Genome Characterisation and Genotyping of Global WU Polyomavirus Strains. %B Journal of virology %D 2010 %C United States %I American Society for Microbiology %V 84 %N 12 %P 6229-34 %@ 1098-5514 %X Exploration of the genetic diversity of WU polyomavirus (WUV) has been limited in terms of the specimen numbers and particularly the sizes of the genomic fragments analyzed. Using whole-genome sequencing of 48 WUV strains collected in four continents over a 5-year period and 16 publicly available whole-genome sequences, we identified three main WUV clades and five subtypes, provisionally termed Ia, Ib, Ic, II, IIIa, and IIIb. Overall nucleotide variation was low (0 to 1.2%). The discriminatory power of the previous VP2 fragment typing method was found to be limited, and a new, larger genotyping region within the VP2/1 interface was proposed. %Z FOR Codes: 1108 1109 %0 Journal Article %~ PubMed %A Playford, Elliott Geoffrey %A Lipman, Jeff %A Kabir, Masrura %A McBryde, Emma S %A Nimmo, Graeme R %A Lau, Anna %A Sorrell, Tania C %T Assessment of clinical risk predictive rules for invasive candidiasis in a prospective multicentre cohort of ICU patients. %B Intensive Care Medicine %D 2009 %C Germany %I Springer %V 35 %N 12 %P 2141-2145 %@ 1432-1238 %X PURPOSE: To assess the generalisability of published clinical risk predictive models for invasive candidiasis in ICU patients. METHODS: The performance characteristics of published clinical risk factor-only and Candida colonisation-only predictive models for invasive candidiasis were assessed in a multicentre cohort of Australian ICU patients. Clinical risk factors and Candida colonisation parameters were collected prospectively from patients. RESULTS: The two clinical risk factor-only predictive models applied to an Australian patient cohort (n = 615) performed less well than in published studies involving derivation populations. Model performance characteristics improved when Candida colonisation parameters were added post-hoc. CONCLUSIONS: Risk predictive models should factor in both clinical risk factors and Candida colonisation parameters. Integrating these models into therapeutic algorithms first requires external validation in different patient populations and settings. %Z FOR Codes: 110309 110310 %0 Journal Article %~ PubMed %A Kong, Fanrong %A Chen, Sharon C A %A Chen, Xiaoyou %A Sintchenko, Vitali %A Halliday, Catriona %A Cai, Lin %A Tong, Zhongsheng %A Lee, Ok Cha %A Sorrell, Tania C %T Assignment of Reference 5'-end 16S rDNA Sequences and Species-Specific Sequence Polymorphisms Improves Species Identification of Nocardia. %B The Open Microbiology Journal %D 2009 %C Netherlands %I Bentham Open %V 3 %N 0 %P 97-105 %@ 1874-2858 %X 16S rDNA sequence analysis is the most accurate method for definitive species identification of nocardiae. However, conflicting results can be found due to sequence errors in gene databases. This study tested the feasibility of species identification of Nocardia by partial (5''-end 606-bp) 16S rDNA sequencing, based on sequence comparison with "reference" sequences of well-annotated strains. This new approach was evaluated using 96 American Type Culture Collection (n=6), and clinical (n=90) Nocardia isolates. Nucleotide sequence-based polymorphisms within species were indicative of "sequence types" for that species. Sequences were compared with those in the GenBank, Bioinformatics Bacteria Identification and Ribosomal Database Project databases. Compared with the reference sequence set, all 96 isolates were correctly identified using the criterion of >/=99% sequence similarity. Seventy-eight (81.3%) were speciated by database comparison; alignment with reference sequences resolved the identity of 14 (15%) isolates whose sequences yielded 100% similarity to sequences in GenBank under >1 species designation. Of 90 clinical isolates, the commonest species was Nocardia nova (33.3%) followed by Nocardia cyriacigeorgica (26.7%). Recently-described or uncommon species included Nocardia veterana (4.4%), Nocarida bejingensis (2.2%) and, Nocardia abscessus and Nocardia arthriditis (each n=1). Nocardia asteroides sensu stricto was rare (n=1). There were nine sequence types of N. nova, three of Nocardia brasiliensis with two each of N. cyriacigeorgica and Nocardia farcinica. Thirteen novel sequences were identified. Alignment of sequences with reference sequences facilitated species identification of Nocardia and allowed delineation of sequence types within species, suggesting that such a barcoding approach can be clinically useful for identification of bacteria. %Z FOR Codes: 60502 110303 60503 %0 Journal Article %~ PubMed %A Brown, Graham V %A Sorrell, Tania C %T Building quality in health--the need for clinical researchers. %B The Medical journal of Australia %D 2009 %C Australia %I Australasian Medical Publishing Company %V 190 %N 11 %P 627-629 %@ 0025-729X %X Integration of research and education into health care delivery leads to improved outcomes and facilitates rapid translation of results into policy and practice. Australia is at great risk of losing the important contribution of clinical research conducted in our public hospital system. This risk is increasing as research and educational training are targeted for expenditure reduction in the current business models of health service delivery, which focus only on short-term outcomes. The Centres of Clinical Research Excellence Scheme--initiated by the National Health and Medical Research Council (NHMRC)--is an excellent step towards redressing this problem, but it cannot succeed in isolation. We must improve and optimise care through promotion of attractive sustainable career pathways to provide strong clinical and translational research capabilities in hospital settings that address current health priorities and new disciplines. Targeted investment in talented people is the greatest long-term contribution that governments can make to guarantee quality in national systems of health. %Z FOR Codes: 111799 %0 Journal Article %~ PubMed %A Chen, S C A %A Marriott, D %A Playford, E G %A Nguyen, Q %A Ellis, D %A Meyer, W %A Sorrell, T C %A Slavin, M %A , the Australian Candidaemia Study %T Candidaemia with uncommon Candida species: predisposing factors, outcome, antifungal susceptibility, and implications for management. %B Clinical microbiology and infection : the official publication of the European Society of Clinical Microbiology and Infectious Diseases %D 2009 %C United Kingdom, Swit %I Wiley-Blackwell Publishing Ltd. %V 15 %N 7 %P 662-9 %@ 1469-0691 %X The risk factors for and clinical features of bloodstream infection with uncommon Candida spp. (species other than C. albicans, C. glabrata, C. parapsilosis, C. tropicals and C. krusei) are incompletely defined. To identify clinical variables associated with these species that might guide management, 57 cases of candidaemia resulting from uncommon Candida spp. were analysed in comparison with 517 episodes of Candida albicans candidaemia (2001-2004). Infection with uncommon Candida spp. (5.3% of candidaemia cases), as compared with C. albicans candidaemia, was significantly more likely to be outpatient-acquired than inpatient-acquired (15 of 57 vs. 65 of 517 episodes, p 0.01). Prior exposure to fluconazole was uncommon (n=1). Candida dubliniensis was the commonest species (n=22, 39%), followed by Candida guilliermondii (n=11, 19%) and Candida lusitaniae (n=7, 12%).C. dubliniensis candidaemia was independently associated with recent intravenous drug use (p 0.01) and chronic liver disease (p 0.03), and infection with species other than C. dubliniensis was independently associated with age<65 years (p 0.02), male sex (p 0.03) and human immunodeficiency virus infection (p 0.05). Presence of sepsis at diagnosis and crude 30-day mortality rates were similar for C. dubliniensis-related, non-C. dubliniensis-related and C. albicans-related candidaemia. Haematological malignancy was the commonest predisposing factor in C. guilliermondii (n=3, 27%) and C. lusitaniae (n=3, 43%) candidaemia. The 30-day mortality rate of C. lusitaniae candidaemia was higher than the overall death rate for all uncommon Candida spp. (42.9% vs. 25%, p not significant). All isolates were susceptible to amphotericin B, voriconazole, posaconazole, and caspofungin; five strains (9%) had fluconazole MIC values of 16-32 mg/L. Candidaemia due to uncommon Candida spp. is emerging among hospital outpatients; certain clinical variables may assist in recognition of this entity. %Z FOR Codes: 1108 1108 1108 %0 Journal Article %~ PubMed %A van Hal, S J %A Marriott, D J E %A Chen, S C A %A Nguyen, Q %A Sorrell, T C %A Ellis, D H %A Slavin, M A %A , Australian Candidaemia Study %T Candidemia following solid organ transplantation in the era of antifungal prophylaxis: the Australian experience. %B Transplant Infectious Disease %D 2009 %C Denmark, United Stat %I Wiley-Blackwell Munksgaard %V 11 %N 2 %P 122-127 %@ 1399-3062 %X Solid organ transplant (SOT) recipients have high rates of invasive fungal infections, with Candida species the most commonly isolated fungi. The aim of this study was to identify differences between incidence rates, risk factors, clinical presentations, and outcomes of candidemia in SOT recipients and non-SOT patients. Data from the multicenter prospective Australian Candidaemia Study were examined. From August 2001 to July 2004, 24 episodes (2.2%; 24/1068) of candidemia were identified in SOT recipients. During this period, the numbers of transplanted organs included liver (n=455), kidney (n=1605), single lung (n=57), bilateral lung (n=183), heart and lung (n=18), heart (n=157), and pancreas (n=62). The overall annual estimated incidence of candidemia in SOT recipients was higher (3 per 1000 transplant admissions) than in non-SOT patients (incidence 0.21 per 1000 admissions; P<0.001). The incidence and timing of candidemia post transplant was influenced by the transplanted organ type, with the majority of episodes (n=14, 54%) occurring >6 months after renal transplantation. Risk factors for candidemia in the month preceding diagnosis were similar to non-SOT recipients except for corticosteroid therapy (P<0.001). Antifungal prophylaxis did not select for more resistant or non-albicans Candida species in the SOT group. The 30-day all-cause mortality was similar to non-SOT patients with candidemia and remains high at 21%. All deaths in SOT recipients occurred early (within 5 days of diagnosis), underlining a need for better diagnostic tests, targeted prevention, and early treatment strategies. %Z FOR Codes: 1115 %0 Journal Article %~ PubMed %A Gordon-Thomson, C %A Kumari, A %A Tomkins, L %A Holford, P %A Djordjevic, J T %A Wright, L C %A Sorrell, T C %A Moore, G P M %T Chitotriosidase and gene therapy for fungal infections. %B Cellular and molecular life sciences %D 2009 %C Switzerland %I Birkhaeuser Verlag AG %V 66 %N 6 %P 1116-1125 %@ 1420-9071 %X Chitotriosidase secreted by activated human macrophages has been implicated in the defence against chitin-bearing pathogens. The antifungal properties of human chitotriosidase were investigated here following retroviral vector-mediated gene transfer of the open reading frame of the chitotriosidase gene into Chinese hamster ovary cells. A chitinase assay confirmed that the engineered cells secreted recombinant chitotriosidase constitutively. Two dimensional gel electrophoresis and western blotting indicated that the recombinant protein is the major, chitin-binding, fifty kilodalton isoform. Culture medium conditioned by the transduced cells inhibited growth of isolates of Aspergillus niger, Candida albicans and Cryptococcus neoformans. Furthermore, longevity was significantly increased in a mouse model of cryptococcosis when cells transduced with the chitotriosidase gene and encapsulated in alginate microspheres were implanted subcutaneously in the animals. Engraftment of microcapsules containing cells transduced with the chitotriosidase gene has the potential to combat infections caused by chitinous pathogens through the prolonged delivery of recombinant chitotriosidase. %Z FOR Codes: 100401 %0 Journal Article %~ PubMed %A Lau, Anna %A Chen, Sharon %A Sleiman, Sue %A Sorrell, Tania %T Current status and future perspectives on molecular and serological methods in diagnostic mycology. %B Future Microbiology %D 2009 %C United Kingdom %I Future Medicine Ltd. %V 4 %N 9 %P 1185-1222 %@ 1746-0921 %X Invasive fungal infections are an important cause of infectious morbidity. Nonculture-based methods are increasingly used for rapid, accurate diagnosis to improve patient outcomes. New and existing DNA amplification platforms have high sensitivity and specificity for direct detection and identification of fungi in clinical specimens. Since laboratories are increasingly reliant on DNA sequencing for fungal identification, measures to improve sequence interpretation should support validation of reference isolates and quality control in public gene repositories. Novel technologies (e.g., isothermal and PNA FISH methods), platforms enabling high-throughput analyses (e.g., DNA microarrays and Luminex xMAP) and/or commercial PCR assays warrant further evaluation for routine diagnostic use. Notwithstanding the advantages of molecular tests, serological assays remain clinically useful for patient management. The serum Aspergillus galactomannan test has been incorporated into diagnostic algorithms of invasive aspergillosis. Both the galactomannan and the serum beta-D-glucan test have value for diagnosing infection and monitoring therapeutic response. %Z FOR Codes: 1108 604 1103 %0 Journal Article %~ PubMed %A Marriott, Deborah %A Playford, E Geoffrey %A Chen, Sharon %A Slavin, Monica %A Nguyen, Quoc %A Ellis, David %A Sorrell, Tania %A , the Australian Candidaemia Study %T Determinants of mortality in non-neutropenic ICU patients with candidaemia. %B Critical care (London, England) %D 2009 %C United Kingdom, Unit %I BioMed Central Ltd. %V 13 %N 4 %P R115 %@ 1466-609X %X Candidaemia in critically-ill intensive care unit (ICU) patients is associated with high crude mortality. Determinants of mortality--particularly those amenable to potential modification--are incompletely defined. %Z FOR Codes: 110399 %0 Book Section %A Sorrell, Tania %A Chen, S C A %T Fungal-Derived Immune Modulating Molecules %B Pathogen-Derived Immunomodulatory Molecules %D 2009 %C United States %I Springer %V %N %P 108-120 %@ 9781441916006 %E Fallon, Padriac G %X %Z FOR Codes: 110799 %0 Journal Article %~ PubMed %A Sorrell, Tania C %A Chen, Sharon C A %T Fungal-derived immune modulating molecules. %B Advances in Experimental Medicine and Biology: Pathogen-Derived Immunomodulatory Molecules %D 2009 %C United States %I Springer New York LLC %V 666 %N %P 108-120 %@ 0065-2598 %X Invasive fungal infections are an increasing clinical problem for which new therapeutic approaches are needed. Understanding the initial interaction between fungi and the host offers potential for development of new drugs or vaccines. It has recently been recognized that like other pathogens, fungi initially interact with the innate immune system via binding between fungus-specific chemical signatures (pattern-associated molecular patterns or PAMPs) and pattern recognition receptors (PRRs) on mononuclear phagocytes. Fungal PAMPs are restricted to complex carbohydrates in the cell wall, including mannoproteins, phospholipomannan, beta-glucans and possibly chitin. These PAMPs bind specifically to two classes of PRR in phagocyte membranes, toll-like receptors and C-lectin-like receptors, through which they initiate signaling responses that culminate in release of pro- and anti-inflammatory cytokines, link the innate immune response with the adaptive immune response and initiate phagocytosis and intracellular killing. Isolated PAMPs have been used to dissect phagocyte responses in vitro and have revealed mechanisms by which host cells can tailor innate immune responses to individual pathogens. The interactions are complex and are yet to be translated into a clear understanding of the roles of the respective PAMPs and PRRs in vivo. Recent advances in this area in relation to the pathogenesis of fungal infections are summarized in this chapter. %Z FOR Codes: 110799 110899 %0 Journal Article %~ PubMed %A Blyth, Christopher C %A Chen, Sharon C A %A Slavin, Monica A %A Serena, Carol %A Nguyen, Quoc %A Marriott, Deborah %A Ellis, David %A Meyer, Wieland %A Sorrell, Tania C %A , Australian Candidemia Study %T Not just little adults: candidemia epidemiology, molecular characterization, and antifungal susceptibility in neonatal and pediatric patients. %B Pediatrics %D 2009 %C United States %I American Academy of Pediatrics %V 123 %N 5 %P 1360-1368 %@ 1098-4275 %X OBJECTIVE: The purpose of this work was to identify differences in incidence, risk factors, microbiology, treatment, and clinical outcome of candidemia in neonates, children, and adults that might impact on management. PATIENTS AND METHODS: Cases of candidemia in Australia were identified prospectively by blood culture surveillance over 3 years. Episodes of candidemia in neonatal, pediatric, and adult age groups were analyzed and compared. RESULTS: Of 1005 incident cases, 33 occurred in neonates, 110 in children, and 862 in adults. The respective annual age-specific incidences were 4.4, 0.9, and 1.8 per 100,000 population. Prematurity and ICU admission were major risk factors in neonates. Hematologic malignancy and neutropenia were significantly more frequent in children than in neonates and adults. Diabetes, renal disease, hemodialysis, and recent surgery were more common in adults. Candidemia was attributed to a vascular access device in 58% of neonates, 70% of children, and 44% of adults. Candida albicans caused approximately 48% of cases in all of the age groups. Candida parapsilosis was significantly more common in neonates and children (42% and 38% vs 15%). Candida glabrata was infrequent in neonates and children (9% and 3% vs 17%). Significantly more isolates from children were susceptible to fluconazole compared with those from adults (95% vs 75%). Fluconazole-resistant candidal isolates were infrequent in all of the age groups. Neonates and children were more likely to receive amphotericin B compared with adults. Adults were more likely to receive fluconazole. Survival rates at 30 days were 78% in neonates, 90% in children, and 70% in adults. CONCLUSIONS: This study identifies significant differences in candidemia in neonates, children, and adults. Neonatologists and pediatricians must consider age-specific differences when interpreting adult studies and developing treatment and prevention guidelines. %Z FOR Codes: 1402 %0 Journal Article %~ PubMed %A Ferguson, Patricia E %A Sorrell, Tania C %A Bradstock, Kenneth F %A Carr, Peter %A Gilroy, Nicole M %T Parainfluenza Virus Type 3 Pneumonia in Bone Marrow Transplant Recipients: Multiple Small Nodules in High-Resolution Lung Computed Tomography Scans Provide a Radiological Clue to Diagnosis. %B Clinical Infectious Diseases %D 2009 %C United States %I University of Chicago Press %V 48 %N 1 %P 905–909 %@ 1058-4838 %X We report the findings of high-resolution chest computed tomography of 6 hematopoietic stem cell transplant recipients with parainfluenza virus type 3 pneumonia who were not infected with any other pathogens. All patients had multiple small nodules (diameter, <5 mm) without cavitation in a peribronchial distribution. Changes preceded microbiological diagnosis in 4 of 6 cases. %Z FOR Codes: 110309 %0 Journal Article %~ PubMed %A Heath, C H %A Slavin, M A %A Sorrell, T C %A Handke, R %A Harun, A %A Phillips, M %A Nguyen, Q %A Delhaes, L %A Ellis, D %A Meyer, W %A Chen, S C A %A , Australian Scedosporium Study Group %T Population-based surveillance for scedosporiosis in Australia: epidemiology, disease manifestations and emergence of Scedosporium aurantiacum infection. %B Clinical Microbiology and Infection %D 2009 %C United Kingdom, Swit %I Wiley-Blackwell Publishing Ltd. %V 15 %N 7 %P 689-693 %@ 1469-0691 %X Australia-wide population-based surveillance for scedosporiosis identified 180 cases, with 118 (65.6%) cases of colonization and 62 (34.4%) cases of infection. Predisposing factors for isolation of Scedosporium spp. included chronic lung disease in 37.8% and malignancy in 21.7% of cases. Predictors of invasive disease (n=62) included haematological stem cell transplantation (n=7), leukaemia (n=16) and diabetes mellitus (n=8). Of 183 phenotypically-speciated isolates, 75 (41%) were Scedosporium prolificans (risk factors: haematologic cancer (n=17), neutropaenia (n=14)) and 108 (59%) had Scedosporium apiospermum/Pseudallescheria boydii phenotype [risk factor: diabetes (n=15)]. Scedosporium prolificans (p 0.01) and leukaemia (p 0.03) independently predicted death. Epidemiological and antifungal susceptibility profiles of Scedosporium aurantiacum (prevalence>or=15.8%) and S. apiospermum were similar. No patient with S. aurantiacum infection (n=6) died. This is the first description of clinical features associated with S. aurantiacum. %Z FOR Codes: 111706 110801 %0 Journal Article %~ PubMed %A Wang, Huiping %A Kong, Fanrong %A Sorrell, Tania C %A Wang, Bin %A McNicholas, Paul %A Pantarat, Namfon %A Ellis, David %A Xiao, Meng %A Widmer, Fred %A Chen, Sharon Ca %T Rapid detection of ERG11 gene mutations in clinical Candida albicans isolates with reduced susceptibility to fluconazole by rolling circle amplification and DNA sequencing. %B BMC Microbiology %D 2009 %C United Kingdom %I BioMed Central Ltd. %V 9 %N %P 167 %@ 1471-2180 %X BACKGROUND: Amino acid substitutions in the target enzyme Erg11p of azole antifungals contribute to clinically-relevant azole resistance in Candida albicans. A simple molecular method for rapid detection of ERG11 gene mutations would be an advantage as a screening tool to identify potentially-resistant strains and to track their movement. To complement DNA sequencing, we developed a padlock probe and rolling circle amplification (RCA)-based method to detect a series of mutations in the C. albicans ERG11 gene using "reference" azole-resistant isolates with known mutations. The method was then used to estimate the frequency of ERG11 mutations and their type in 25 Australian clinical C. albicans isolates with reduced susceptibility to fluconazole and in 23 fluconazole-susceptible isolates. RCA results were compared DNA sequencing. RESULTS: The RCA assay correctly identified all ERG11 mutations in eight "reference" C. albicans isolates. When applied to 48 test strains, the RCA method showed 100% agreement with DNA sequencing where an ERG11 mutation-specific probe was used. Of 20 different missense mutations detected by sequencing in 24 of 25 (96%) isolates with reduced fluconazole susceptibility, 16 were detected by RCA. Five missense mutations were detected by both methods in 18 of 23 (78%) fluconazole-susceptible strains. DNA sequencing revealed that mutations in non-susceptible isolates were all due to homozygous nucleotide changes. With the exception of the mutations leading to amino acid substitution E266D, those in fluconazole-susceptible strains were heterozygous. Amino acid substitutions common to both sets of isolates were D116E, E266D, K128T, V437I and V488I. Substitutions unique to isolates with reduced fluconazole susceptibility were G464 S (n = 4 isolates), G448E (n = 3), G307S (n = 3), K143R (n = 3) and Y123H, S405F and R467K (each n = 1). DNA sequencing revealed a novel substitution, G450V, in one isolate. CONCLUSION: The sensitive RCA assay described here is a simple, robust and rapid (2 h) method for the detection of ERG11 polymorphisms. It showed excellent concordance with ERG11 sequencing and is a potentially valuable tool to track the emergence and spread of azole-resistant C. albicans and to study the epidemiology of ERG11 mutations. The RCA method is applicable to the study of azole resistance in other fungi. %Z FOR Codes: 110801 %0 Journal Article %~ PubMed %A Himmelreich, Uwe %A Malik, Richard %A Kühn, Till %A Daniel, Heide-Marie %A Somorjai, Ray L %A Dolenko, Brion %A Sorrell, Tania C %T Rapid etiological classification of meningitis by NMR spectroscopy based on metabolite profiles and host response. %B PLoS One %D 2009 %C United States %I Public Library of Science %V 4 %N 4 %P e5328 %@ 1932-6203 %X Bacterial meningitis is an acute disease with high mortality that is reduced by early treatment. Identification of the causative microorganism by culture is sensitive but slow. Large volumes of cerebrospinal fluid (CSF) are required to maximise sensitivity and establish a provisional diagnosis. We have utilised nuclear magnetic resonance (NMR) spectroscopy to rapidly characterise the biochemical profile of CSF from normal rats and animals with pneumococcal or cryptococcal meningitis. Use of a miniaturised capillary NMR system overcame limitations caused by small CSF volumes and low metabolite concentrations. The analysis of the complex NMR spectroscopic data by a supervised statistical classification strategy included major, minor and unidentified metabolites. Reproducible spectral profiles were generated within less than three minutes, and revealed differences in the relative amounts of glucose, lactate, citrate, amino acid residues, acetate and polyols in the three groups. Contributions from microbial metabolism and inflammatory cells were evident. The computerised statistical classification strategy is based on both major metabolites and minor, partially unidentified metabolites. This data analysis proved highly specific for diagnosis (100% specificity in the final validation set), provided those with visible blood contamination were excluded from analysis; 6-8% of samples were classified as indeterminate. This proof of principle study suggests that a rapid etiologic diagnosis of meningitis is possible without prior culture. The method can be fully automated and avoids delays due to processing and selective identification of specific pathogens that are inherent in DNA-based techniques. %Z FOR Codes: 110309 %0 Journal Article %~ PubMed %A Kesson, Alison M %A Bellemore, Michael C %A O'Mara, Timothy J %A Ellis, David H %A Sorrell, Tania C %T Scedosporium prolificans osteomyelitis in an immunocompetent child treated with a novel agent, hexadecylphospocholine (miltefosine), in combination with terbinafine and voriconazole: a case report. %B Clinical Infectious Diseases %D 2009 %C United States %I University of Chicago Press %V 48 %N 9 %P 1257-1261 %@ 1058-4838 %X We describe an 8-year-old girl who sustained multiple compound fractures in an accident involving agricultural equipment. She developed Scedosporium prolificans osteomyelitis of the pelvis, septic arthritis of the hip, and myositis of adjacent muscles. The infection progressed, despite extensive surgical debridement and joint washouts with 0.2% polyhexamethylene biguanide; antifungal therapy with caspofungin, terbinafine, and voriconazole; and adjunctive therapy with interferon-gamma. Gradual resolution was achieved after the addition of a novel agent, hexadecylphospocholine (miltefosine), and the continuation of terbinafine and voriconazole. This is the first report of the use of miltefosine as an antifungal agent in the management of severe infection with S. prolificans. %Z FOR Codes: 110802 110309 111403 %0 Journal Article %~ PubMed %A Obando, Daniel %A Pantarat, Namfon %A Handke, Rosemary %A Koda, Yasuko %A Widmer, Fred %A Djordjevic, Julianne T %A Ellis, David H %A Sorrell, Tania C %A Jolliffe, Katrina A %T Synthesis, antifungal, haemolytic and cytotoxic activities of a series of bis(alkylpyridinium)alkanes. %B Bioorganic & Medicinal Chemistry %D 2009 %C United Kingdom %I Pergamon %V 17 %N 17 %P 6329-6339 %@ 0968-0896 %X A series of bis(alkylpyridinium)alkanes with a twelve carbon spacer between the positive charges was synthesised and their antifungal activity has been investigated. Compounds with 2-pentyl, 4-pentyl, 4-hexyl, 4-octyl, 4-propylbenzene, 3,4-dipentyl, 4-(5''-nonyl) and 3-methyl,4-pentyl head groups were the most potent antifungal agents with MICs in the range of 1.4-2.7 microM against reference strains of both Cryptococcus neoformans and Candida albicans. %Z FOR Codes: 110199 %0 Journal Article %~ PubMed %A Ngamskulrungroj, Popchai %A Sorrell, Tania %A Chindamporn, Ariya %A Chaiprasert, Angkana %A Poonwan, Natthiwan %A Meyer, Wieland %T Association between fertility and molecular sub-type of global isolates of Cryptococcus gattii molecular type VGII. %B Medical mycology : official publication of the International Society for Human and Animal Mycology %D 2008 %C United Kingdom %I Informa Healthcare %V 46 %N 0 %P 665-73 %@ 1369-3786 %X The basidiomycetous yeast Cryptococcus gattii, is a primary pathogen which causes disease in apparently healthy humans and a wide range of animals. Recently, an outbreak of cryptococcosis caused by a previously uncommon genotype of C. gattii, VGII, emerged on Vancouver Island, British Columbia, Canada. Two pathogenic sub-types of VGII (designated VGIIa and VGIIb) were identified among these isolates. All of the isolates proved to be mating type alpha and had exceptionally high sporulation capacity. The common subtype, VGIIa, was more virulent than VGIIb in mice, suggesting a linkage between subtype and fertility/virulence. To test this hypothesis, we compared the fertility of 91 isolates from the Vancouver Island outbreak with that of 72 VGII isolates selected globally. Of all isolates, 69.94% were found to be fertile and exhibited clamp connections and basidiospores. The Vancouver isolates showed a high fertility rate of 84.2% as compared to only 29% of the 21 Australian isolates investigated. Mating type alpha strains were more fertile (72.79%) than mating type a (43.75%) (p<0.022). Amongst the two subtypes of VGII a much higher proportion of VGIIa (91.7%) than VGIIb (33.3%) was fertile (p<0.001). These results suggest that there is a clear correlation between the VGII subtypes of C. gattii and their mating/fertility. Further in vitro and in vivo investigations of more strains and congenic pairs are warranted. %Z FOR Codes: 1108 %0 Journal Article %~ PubMed %A Playford, E %A Marriott, Deborah %A Nguyen, Quoc %A Chen, Sharon %A Ellis, David %A Slavin, Monica %A Sorrell, Tania %T Candidemia in nonneutropenic critically Ill patients: Risk factors for non-albicans Candida spp. %B Critical care medicine %D 2008 %C United States %I Lippincott Williams and Wilkins %V 36 %N 7 %P 2034-9 %@ 0090-3493 %X The objective of this study was to determine the clinical features associated with candidemia caused by non-albicans Candida spp. and with potentially fluconazole-resistant Candida spp. (C. glabrata and C. krusei) among candidemic intensive care unit patients. %Z FOR Codes: 110899 %0 Journal Article %~ PubMed %A Lau, Anna %A Sorrell, Tania C %A Lee, Okcha %A Stanley, Keith %A Halliday, Catriona %T Colony Multiplex-Tandem PCR for Rapid, Accurate Identification of Fungal Cultures. %B Journal of clinical microbiology %D 2008 %C United States %I American Society for Microbiology %V 46 %N 12 %P 4058-60 %@ 0095-1137 %X We developed a multiplex tandem PCR (MT-PCR) assay for the rapid identification of 16 fungi directly from culture. MT-PCR results were concordant with phenotypic identification for all cultures studied (n = 183). The colony MT-PCR assay was rapid (<2 h), sensitive, and specific in identifying fungal pathogens directly from primary isolation plates. %Z FOR Codes: 110899 %0 Journal Article %~ PubMed %A Delhaes, Laurence %A Harun, Azian %A Chen, Sharon C A %A Nguyen, Quoc %A Slavin, Monica %A Heath, Christopher H %A Maszewska, Krystyna %A Halliday, Catriona %A Robert, Vincent %A Sorrell, Tania C %A Auscedo Study Group, The Australian Scedosporium %A Meyer, Wieland %T Molecular typing of Australian Scedosporium isolates showing genetic variability and numerous S. aurantiacum. %B Emerging Infectious Diseases %D 2008 %C United States %I Center Disease Control %V 14 %N 2 %P 282-290 %@ 1080-6040 %X One hundred clinical isolates from a prospective nationwide study of scedosporiosis in Australia (2003-2005) and 46 additional isolates were genotyped by internal transcribed spacer-restriction fragment length polymorphism (ITS-RFLP) analysis, ITS sequencing, and M13 PCR fingerprinting. ITS-RFLP and PCR fingerprinting identified 3 distinct genetic groups. The first group corresponded to Scedosporium prolificans (n = 83), and the other 2 comprised isolates previously identified as S. apiospermum: one of these corresponded to S. apiospermum (n = 33) and the other to the newly described species S. aurantiacum (n = 30). Intraspecies variation was highest for S. apiospermum (58%), followed by S. prolificans (45%) and S. aurantiacum (28%) as determined by PCR fingerprinting. ITS sequence variation of 2.2% was observed among S. apiospermum isolates. No correlation was found between genotype of strains and their geographic origin, body site from which they were cultured, or colonization versus invasive disease. Twelve S. prolificans isolates from 2 suspected case clusters were examined by amplified fragment length polymorphism analysis. No specific clusters were confirmed. %Z FOR Codes: 110899 %0 Journal Article %~ PubMed %A Lau, Anna %A Sorrell, Tania %A Chen, Sharon %A Stanley, Keith %A Iredell, Jonathan %A Halliday, Catriona %T Multiplex-Tandem PCR: A Novel Platform for the Rapid Detection and Identification of Fungal Pathogens from Blood Culture Specimens. %B Journal of clinical microbiology %D 2008 %C United States %I American Society of Microbiology %V 46 %N 9 %P 3021-7 %@ 1098-660X %X We describe the first development and evaluation of a rapid multiplex tandem PCR (MT-PCR) assay for the detection and identification of fungi directly from blood culture specimens that have been flagged as positive. The assay uses a short-cycle multiplex amplification, followed by 12 simultaneous PCRs which target the fungal internal transcribed spacer 1 (ITS1) and ITS2 region, elongation factor 1-alpha (EF1-alpha), and beta-tubulin genes to identify 11 fungal pathogens: Candida albicans, Candida dubliniensis, Candida glabrata, Candida guilliermondii, Candida krusei, Candida parapsilosis complex, Candida tropicalis, Cryptococcus neoformans complex, Fusarium solani, Fusarium species, and Scedosporium prolificans. The presence or absence of a fungal target was confirmed by melting curve analysis. Identification by MT-PCR correlated with culture-based identification for 44 (100%) patients. No cross-reactivity was detected in 200 blood culture specimens that contained bacteria or in 30 blood cultures without microorganisms. Fungi were correctly identified in five specimens with bacterial coinfection and in blood culture samples that were seeded with a mixture of yeast cells. The MT-PCR assay was able to provide rapid (<2 h), sensitive, and specific simultaneous detection and identification of fungal pathogens directly from blood culture specimens. %Z FOR Codes: 110899 %0 Journal Article %A Sorrell, Tania %A Himmelreich, U %T NMR in mycology %B Current Fungal Infection Reports %D 2008 %C United States %I Current Medicine Group LLC %V 2 %N %P 149-156 %@ 1936-3761 %X %Z FOR Codes: 110899 %0 Journal Article %~ PubMed %A Zhou, Xiaoyong %A Kong, Fanrong %A Sorrell, Tania C %A Wang, Hui %A Duan, Yiqun %A Chen, Sharon C A %T Practical method for detection and identification of Candida, Aspergillus and Scedosporium using rolling circle amplification (RCA). %B Journal of clinical microbiology %D 2008 %C United States %I American Society for Microbiology %V 46 %N 7 %P 2423-7 %@ 0095-1137 %X A sensitive rolling-circle amplification (RCA)-based method utilizing species-specific padlock probes targeted to the internal transcribed spacer 2 region of the fungal ribosomal DNA gene complex was developed. The assay was rapid (2 hours) and specific. Of 28 fungal isolates (16 of Candida, six of Aspergillus, and six of Scedosporium spp.), all were all identified correctly. %Z FOR Codes: 110899 %0 Journal Article %~ PubMed %A Kong, Fanrong %A Tong, Zhongsheng %A Chen, Xiaoyou %A Sorrell, Tania %A Wang, Bin %A Wu, Qixuan %A Ellis, David %A Chen, Sharon %T Rapid identification and differentiation of Trichophyton species, based on sequence polymorphisms of the ribosomal internal transcribed spacer regions, by rolling-circle amplification. %B Journal of Clinical Microbiology %D 2008 %C United States %I American Society for Microbiology %V 46 %N 4 %P 1192-1199 %@ 0095-1137 %X DNA sequencing analyses have demonstrated relatively limited polymorphisms within the fungal internal transcribed spacer (ITS) regions among Trichophyton spp. We sequenced the ITS region (ITS1, 5.8S, and ITS2) for 42 dermatophytes belonging to seven species (Trichophyton rubrum, T. mentagrophytes, T. soudanense, T. tonsurans, Epidermophyton floccosum, Microsporum canis, and M. gypseum) and developed a novel padlock probe and rolling-circle amplification (RCA)-based method for identification of single nucleotide polymorphisms (SNPs) that could be exploited to differentiate between Trichophyton spp. Sequencing results demonstrated intraspecies genetic variation for T. tonsurans, T. mentagrophytes, and T. soudanense but not T. rubrum. Signature sets of SNPs between T. rubrum and T. soudanense (4-bp difference) and T. violaceum and T. soudanense (3-bp difference) were identified. The RCA assay correctly identified five Trichophyton species. Although the use of two "group-specific" probes targeting both the ITS1 and the ITS2 regions were required to identify T. soudanense, the other species were identified by single ITS1- or ITS2-targeted species-specific probes. There was good agreement between ITS sequencing and the RCA assay. Despite limited genetic variation between Trichophyton spp., the sensitive, specific RCA-based SNP detection assay showed potential as a simple, reproducible method for the rapid (2-h) identification of Trichophyton spp. %Z FOR Codes: 110899 %0 Journal Article %~ PubMed %A Branley, J M %A Roy, B %A Dwyer, D E %A Sorrell, T C %T Real-time PCR detection and quantitation of Chlamydophila psittaci in human and avian specimens from a veterinary clinic cluster. %B European journal of clinical microbiology & infectious diseases %D 2008 %C Germany %I Springer %V 27 %N 4 %P 269-273 %@ 0934-9723 %X We report three cases of psittacosis in staff working in a veterinary surgery, which was related to exposure to a sick, wild psittacine bird. Chlamydial genus- and chlamydial species-specific DNA was detected in clinical specimens, including throat swabs, whole blood and urine. The organism load was quantified by real-time PCR (RT-PCR), which revealed 10(5)-fold more organisms in conjunctival swabs from the source bird than in the human samples. One clinic attendant was infected despite using personal protective equipment when handling the bird. This is the first report of PCR analyses of blood and urine samples being used to diagnose human psittacosis, and the first time that the organism load in humans has been compared to that of the infecting bird. %Z FOR Codes: 110309 %0 Journal Article %~ PubMed %A Thursky, K A %A Playford, E G %A Seymour, J F %A Sorrell, T C %A Ellis, D H %A Guy, S D %A Gilroy, N %A Chu, J %A Shaw, D R %T Recommendations for the treatment of established fungal infections. %B Internal medicine journal %D 2008 %C Australia %I Blackwell Publishing Asia %V 38 %N 6b %P 496-520 %@ 1444-0903 %X Evidence-based guidelines for the treatment of established fungal infections in the adult haematology/oncology setting were developed by a national consensus working group representing clinicians, pharmacists and microbiologists. These updated guidelines replace the previous guidelines published in the Internal Medicine Journal by Slavin et al. in 2004. The guidelines are pathogen-specific and cover the treatment of the most common fungal infections including candidiasis, aspergillosis, cryptococcosis, zygomycosis, fusariosis, scedosporiosis, and dermatophytosis. Recommendations are provided for management of refractory disease or salvage therapies, and special sites of infections such as the cerebral nervous system and the eye. Because of the widespread use newer broad-spectrum triazoles in prophylaxis and empiric therapy, these guidelines should be implemented in concert with the updated prophylaxis and empiric therapy guidelines published by this group. %Z FOR Codes: 110309 %0 Journal Article %~ PubMed %A De Pauw, Ben %A Walsh, Thomas J %A Donnelly, J Peter %A Stevens, David A %A Edwards, John E %A Calandra, Thierry %A Pappas, Peter G %A Maertens, Johan %A Lortholary, Olivier %A Kauffman, Carol A %A Denning, David W %A Patterson, Thomas F %A Maschmeyer, Georg %A Bille, Jacques %A Dismukes, William E %A Herbrecht, Raoul %A Hope, William W %A Kibbler, Christopher C %A Kullberg, Bart Jan %A Marr, Kieren A %A Muņoz, Patricia %A Odds, Frank C %A Perfect, John R %A Restrepo, Angela %A Ruhnke, Markus %A Segal, Brahm H %A Sobel, Jack D %A Sorrell, Tania C %A Viscoli, Claudio %A Wingard, John R %A Zaoutis, Theoklis %A Bennett, John E %A , European Organization for Research and Treatment of Cancer/Invasive Fungal Infections Cooperative Group %A , National Institute of Allergy and Infectious Diseases Mycoses Study Group (EORTC/MSG) Consensus Group %T Revised definitions of invasive fungal disease from the European Organization for Research and Treatment of Cancer/Invasive Fungal Infections Cooperative Group and the National Institute of Allergy and Infectious Diseases Mycoses Study Group (EORTC/MSG) Consensus Group. %B Clinical infectious diseases %D 2008 %C United States %I University of Chicago Press %V 46 %N 12 %P 1813-1821 %@ 1058-4838 %X BACKGROUND: Invasive fungal diseases are important causes of morbidity and mortality. Clarity and uniformity in defining these infections are important factors in improving the quality of clinical studies. A standard set of definitions strengthens the consistency and reproducibility of such studies. METHODS: After the introduction of the original European Organization for Research and Treatment of Cancer/Invasive Fungal Infections Cooperative Group and the National Institute of Allergy and Infectious Diseases Mycoses Study Group (EORTC/MSG) Consensus Group definitions, advances in diagnostic technology and the recognition of areas in need of improvement led to a revision of this document. The revision process started with a meeting of participants in 2003, to decide on the process and to draft the proposal. This was followed by several rounds of consultation until a final draft was approved in 2005. This was made available for 6 months to allow public comment, and then the manuscript was prepared and approved. RESULTS: The revised definitions retain the original classifications of "proven," "probable," and "possible" invasive fungal disease, but the definition of "probable" has been expanded, whereas the scope of the category "possible" has been diminished. The category of proven invasive fungal disease can apply to any patient, regardless of whether the patient is immunocompromised, whereas the probable and possible categories are proposed for immunocompromised patients only. CONCLUSIONS: These revised definitions of invasive fungal disease are intended to advance clinical and epidemiological research and may serve as a useful model for defining other infections in high-risk patients. %Z FOR Codes: 110309 %0 Journal Article %~ PubMed %A Chayakulkeeree, Methee %A Sorrell, Tania C %A Siafakas, A Rosemary %A Wilson, Christabel F %A Pantarat, Namfon %A Gerik, Kimberly J %A Boadle, Ross %A Djordjevic, Julianne T %T Role and Mechanism of Phosphatidylinositol-Specific Phospholipase C in Survival and Virulence of Cryptococcus neoformans. %B Molecular microbiology %D 2008 %C United Kingdom %I Wiley-Blackwell Publishing Ltd. %V 69 %N 4 %P 809-26 %@ 0950-382X %X Phospholipase B1 (Plb1) is secreted after release from its glycosylphosphatidylinositol anchor and is implicated in initiation and dissemination of infection of the pathogenic fungus, Cryptococcus neoformans. To investigate the role of phosphatidylinositol-specific phospholipase C (PI-PLC) in Plb1 secretion, we identified two putative PI-PLC-encoding genes in C. neoformans var. grubii (PLC1 and PLC2), and created Deltaplc1 and Deltaplc2 deletion mutants. In Deltaplc1, which expressed less PI-PLC activity than wild type (WT), three major cryptococcal virulence traits, Plb1 secretion, melanin production and growth at host temperature (37 degrees C) were abolished and absence of Plb1 secretion coincided with Plb1 accumulation in plasma membranes. In addition, Deltaplc1 cell walls were defective, as indicated by cell clumping and irregular morphology, slower growth and an inability to activate mitogen-activated protein kinase (MAPK) in the presence of cell wall-perturbing agents. In contrast to Deltaplc2, which was as virulent as WT, Deltaplc1 was avirulent in mice and exhibited attenuated killing of Caenorhabditis elegans at 25 degrees C, demonstrating that mechanism(s) independent of the 37 degrees C growth defect contribute to the virulence composite. We conclude that Plc1 is a central regulator of cryptococcal virulence, acting through the protein kinase C/MAPK pathway, that it regulates release of Plb1 from the plasma membrane and is a candidate antifungal drug target. %Z FOR Codes: 110899 %0 Journal Article %~ PubMed %A Chen, Sharon C A %A Sorrell, Tania C %T Antifungal agents %B The Medical journal of Australia %D 2007 %C Australia %I Australasian Medical Publishing Company Pty. Ltd. %V 187 %N 7 %P 404-409 %@ 1326-5377 %X The four main classes of antifungal drugs are the polyenes, azoles, allylamines and echinocandins. Clinically useful "older" agents include topical azole formulations (for superficial yeast and dermatophyte infections), first-generation triazoles (fluconazole and itraconazole, for a range of superficial and invasive fungal infections), amphotericin B formulations (for a broad range of invasive fungal infections) and terbinafine (for dermatophyte infections). Clinically important "newer" agents include members of the echinocandin class (eg, caspofungin) and second-generation triazoles (eg, voriconazole and posaconazole). Voriconazole and posaconazole have broad-spectrum activity against yeasts and moulds, including Aspergillus species. Posaconazole is the only azole drug with activity against zygomycete fungi. Caspofungin and the other echinocandins are effective in treating Candida and Aspergillus infections. The azoles are relatively safe, but clinicians should be aware of drug-drug interactions and adverse effects, including visual disturbances (with voriconazole), elevations in liver transaminase levels, and skin rashes. Caspofungin has minimal adverse effects. Combination antifungal therapy may be appropriate in selected patients with invasive fungal infections, but is empiric and driven by individual physician practice. %Z FOR Codes: 110309 %0 Journal Article %~ PubMed %A Siafakas, A Rosemary %A Sorrell, Tania C %A Wright, Lesley C %A Wilson, Christabel %A Larsen, Michelle %A Boadle, Ross %A Williamson, Peter R %A Djordjevic, Julianne T %T Cell wall-linked cryptococcal phospholipase B1 is a source of secreted enzyme and a determinant of cell wall integrity. %B The Journal of biological chemistry %D 2007 %C Bethesda MD, USA %I The American Society for Biochemistry and Molecula %V 282 %N 52 %P 37508-14 %@ 1083-351X %X Phospholipase B (Plb1) is secreted by pathogenic fungi and is a proven virulence determinant in Cryptococcus neoformans. Cell-associated Plb1 is presumptively involved in fungal membrane biogenesis and remodelling. We have also identified it in cryptococcal cell walls. Motif scanning programs predict that Plb1 is attached to cryptococcal membranes via a glycosylphosphatidylinositol (GPI) anchor, which could regulate Plb1 export and secretion. A functional GPI anchor was identified in cell-associated Plb1 by (G)PI-specific phospholipase C (PLC)-induced release of Plb1 from strain H99 membrane rafts and inhibition of GPI anchor synthesis by YW3548, which prevented Plb1 secretion and transport to membranes and cell walls. Plb1 containing beta-1,6-linked glucan was released from H99 (wild-type strain) cell walls by beta-1,3 glucanase, consistent with covalent attachment of Plb1 via beta-1,6-linked glucans to beta-1,3-linked glucan in the central scaffold of the wall. Naturally secreted Plb1 also contained beta-1,6-linked glucan, confirming that it originated from the cell wall. Plb1 maintains cell wall integrity because a H99 deletion mutant, DeltaPLB1, exhibited a morphological defect and was more susceptible than H99 to cell wall disruption by SDS and Congo red. Growth of DeltaPLB1 was unaffected by caffeine, excluding an effect of Plb1 on cell wall biogenesis-related signaling pathways. Environmental (heat) stress caused Plb1 accumulation in cell walls, with loss from membranes and reduced secretion, further supporting the importance of Plb1 in cell wall integrity. This is the first demonstration that Plb1 contributes to fungal survival by maintaining cell wall integrity and that the cell wall is a source of secreted enzyme. %Z FOR Codes: 110899 %0 Journal Article %~ PubMed %A Chen, S %A Tong, Z %A Lee, O %A Halliday, C %A Playford, E %A Widmer, F %A Kong, F %A Wu, C %A Sorrell, T %T Clinician response to Candida organisms in the urine of patients attending hospital. %B European journal of clinical microbiology & infectious diseases : official publication of the European Society of Clinical Microbiology %D 2007 %C Germany %I Springer %V 27 %N 0 %P 201-8 %@ 0934-9723 %X The epidemiology of 54 episodes of candiduria with respect to clinical risk factors, species of Candida and physician response to the isolation of Candida in urine were studied in an observational survey over 3 months. Candida spp. were isolated from 4.7% of positive urine cultures. Common predisposing conditions included antibiotic use (74.1%), urinary drainage devices (57.4%), surgery (51.9%), intensive care unit (ICU) or high-dependency care unit (HDU) admission (42.6%) and urinary tract (UT) disease (18.5%). Upper UT infection was uncommon (n = 3). Of 65 Candida isolates, C. albicans predominated (85.2%), followed by C. glabrata (27.8%) and other Candida spp. (6.2%). All isolates were susceptible to fluconazole, itraconazole, voriconazole, amphotericin and caspofungin. Indwelling urinary catheters were removed in 76.2% of episodes. Antifungal therapy was initiated in 33.3% of cases independently of patient symptoms, underlying disease or Candida colony count. Patients in ICU/HDUs were significantly more likely to receive antifungal agents than those outside these units (p < 0.001). Fluconazole was the most common drug prescribed (77.8%). Clearance of candiduria occurred independently of antifungal therapy (p = 0.60). Physicians often did not follow up a positive urine result for Candida. Efforts to increase clinician awareness of current recommendations for managing candiduria and further study to elucidate specific risk factors in defined patient populations are warranted. %Z FOR Codes: 110309 110303 %0 Journal Article %~ PubMed %A Wu, Qi Xuan %A Chen, Sharon C A %A Santangelo, Rosemary T %A Martin, Patricia %A Malik, Richard %A Sorrell, Tania C %T Cryptococcal phospholipase B antigen is not detected in serum of patients infected with Cryptococcus neoformans using a sandwich enzyme-linked immunosorbent assay. %B FEMS yeast research %D 2007 %C United Kingdom %I Blackwell Publishing Ltd. %V 7 %N 3 %P 465-470 %@ 1567-1356 %X Extracellular phospholipase B (PLB) is a virulence determinant of Cryptococcus neoformans and Cryptococcus gattii. In this study, we developed a sensitive enzyme-linked immunosorbent assay (ELISA) for PLB antigen with a detection limit of 3.9 ng mL(-1). PLB was detected in culture supernatants of C. neoformans and C. gattii. PLB, however, was not detected in sera of seven human patients and 10 feline patients with active cryptococcosis. Furthermore, none of five rats with extensive pulmonary C. gattii infection had a positive ELISA test result. In conclusion, cryptococcal PLB could not be detected in serum using a PLB antigen-based ELISA. Despite its sensitivity, this ELISA is of limited diagnostic value. Exploration of further extracellular molecules suitable for serodiagnosis of active cryptococcal infection is warranted. %Z FOR Codes: 110899 %0 Book Section %A Sorrell, Tania %T Detection of Fungal Metabolites %B Diagnosis of Fungal Infections %D 2007 %C United States %I Informa Healthcare %V %N %P 121-132 %@ 9780824729332 %E Maertens, Johan A %E Marr, Kieren A. %X %Z FOR Codes: 110107 %0 Journal Article %~ PubMed %A Plummer, R %A Bodkin, J %A Power, D %A Pantarat, N %A Bubb, W A %A Kuchel, P W %A Sorrell, T C %T Effect of Caspofungin on Metabolite Profiles of Aspergillus Species Determined by Nuclear Magnetic Resonance Spectroscopy. %B Antimicrobial agents and chemotherapy %D 2007 %C USA %I American Society for Microbiology %V 51 %N 11 %P 4077-84 %@ 0066-4804 %X Invasive aspergillosis remains a potentially life-threatening infection, the incidence of which is increasing. Current methods used to determine the susceptibilities of Aspergillus strains to antifungal drugs are often unreliable. Nuclear magnetic resonance (NMR) spectroscopy can identify the metabolic complement of microorganisms while monitoring nutrient utilization from the incubation medium. We used 600-MHz (1)H NMR spectroscopy to monitor the metabolic responses of five Aspergillus species cultured in RPMI 1640-2% glucose-morpholinepropanesulfonate buffer to various concentrations of the antifungal drugs amphotericin B (AMB) and caspofungin. The metabolic endpoint (MEP) was determined from nutrient and metabolite resonances, measured as a function of the drug concentration, and was defined as a > or =50% reduction in nutrient consumption or metabolite production. MICs were evaluated by a modification of Clinical and Laboratory Standards Institute broth microdilution method M27-A, and minimal effective concentrations (MECs) were determined by microscopic examination of fungal hyphae. For AMB, the MEPs coincided with the MICs. For caspofungin, the MEPs agreed with the MECs for several Aspergillus strains, but the effect of drug pressure was more complex for others. Expansion of the MEP definition to include any significant changes in metabolite production resulted in agreement with the MEC in most cases. Paradoxical metabolic responses were observed for several Aspergillus strains at either high or low caspofungin concentrations and for one Aspergillus terreus strain with AMB. NMR spectroscopy proved to be a powerful tool for detecting the subtle effects of drug pressure on fungal metabolism and has the potential to provide an alternative method for determining the susceptibilities of Aspergillus species to antifungal drugs. %Z FOR Codes: %0 Journal Article %~ PubMed %A Tong, Zhongsheng %A Widmer, Fred %A Sorrell, Tania C %A Guse, Zofia %A Jolliffe, Katrina A %A Halliday, Catriona %A Lee, Ok Cha %A Kong, Fanrong %A Wright, Lesley C %A Chen, Sharon C A %T In vitro activities of miltefosine and two novel antifungal biscationic salts against a panel of 77 dermatophytes. %B Antimicrobial agents and chemotherapy %D 2007 %C USA %I American Society for Microbiology %V 51 %N 6 %P 2219-2222 %@ 0066-4804 %X The susceptibilities of 77 dermatophytes to miltefosine (MI), 1,12-bis(4-pentylpyridinium)dodecane (PYR), 1,12-bis(tributylammonium)dodecane (AM), itraconazole (ITC), terbinafine (TRB), and butenafine (BTF) were compared. Geometric mean MICs of TRB, BTF, ITC, MI, PYR, and AM were 0.039, 0.059, 1.718, 0.671, 6.006, and 4.771 microg/ml, respectively. MI was more active than ITC (P < 0.001). %Z FOR Codes: 110899 %0 Journal Article %~ PubMed %A Plummer, R %A Bodkin, J %A Yau, T W %A Power, D %A Pantarat, N %A Larkin, T J %A Szekely, D %A Bubb, W A %A Sorrell, T C %A Kuchel, P W %T Modelling Staphylococcus aureus-induced septicemia using NMR. %B Magnetic resonance in medicine : official journal of the Society of Magnetic Resonance in Medicine / Society of Magnetic Resonance in Medicine %D 2007 %C United States %I John Wiley & Sons, Inc. %V 58 %N 4 %P 656-665 %@ 0740-3194 %X We present a novel NMR-based study of the molecular aspects of the "attack" on human red blood cells (RBCs) by growing bacteria. Staphylococcus aureus expresses virulence factors, including alpha-hemolysin, which contribute to the clinical condition known as septic shock. alpha-Hemolysin is a pore-forming toxin and its secretion increases the permeability of a range of mammalian cell types infected with S. aureus. (31)P NMR spectra of the probe molecules dimethyl methylphosphonate (DMMP) and hypophosphite (HPA) in RBC suspensions show separate intra- and extracellular resonances. These resonances coalesced over time in RBC suspensions inoculated with S. aureus or pure alpha-hemolysin, due to increasing permeability of the RBC membrane. Increased RBC permeability resulted in leakage of intracellular proteins, plus an increase in the exchange rate of the solutes between the intra- and extracellular compartments, both effects contributing to the coalescence of the split peaks. The addition of antibiotics prevented peak coalescence and enabled the minimal inhibitory concentration (MIC) for eight strains of S. aureus to be determined for oxacillin and erythromycin. The MIC values obtained by using (31)P NMR spectroscopy were within one dilution of the MICs obtained using the standard National Committee for Clinical Laboratory Standards (NCCLS) method. The results are encouraging for the use of NMR spectroscopy in clinical microbiology. Magn Reson Med 58:656-665, 2007. (c) 2007 Wiley-Liss, Inc. %Z FOR Codes: %0 Journal Article %~ PubMed %A Zeng, Xianyu %A Kong, Fanrong %A Halliday, Catriona %A Chen, Sharon %A Lau, Anna %A Playford, Geoffrey %A Sorrell, Tania C %T Reverse line blot (RLB) hybridization assay for the identification of medically important fungi from culture and clinical specimens. %B Journal of clinical microbiology %D 2007 %C United States %I American Society of Microbiology %V 45 %N 9 %P 2872-80 %@ 1098-660X %X We evaluated a combined panfungal PCR-reverse line blot (RLB) hybridization assay based on internal transcribed spacer 1 (ITS1) and ITS2 region polymorphisms to identify 159 Candida, Cryptococcus neoformans, and Aspergillus isolates (22 species). Its utility to identify fungal pathogens directly from 27 clinical specimens was also determined. ITS sequence analysis was performed to resolve discrepant identifications or where no RLB result was obtained. Species-specific ITS2- and ITS1-based probes correctly identified 155 of 159 isolates (98%) and 149 (93.7%) isolates, respectively. All strains were unambiguously differentiated with the exception of cross-reactivity between the Candida norvegensis probe and Candida haemulonii DNA product. Species identification of the pathogen was made for all 21 specimens (sensitivity of 100%) where species-specific probes were included in the RLB; however, there was no ITS2 probe-based hybridization signal for two specimens. Results were concordant with the culture results for 18 (85.7%) specimens. The assay was able to provide species identification in the absence of a culture result (two specimens) and to detect mixed infection (one specimen). The results indicate that the RLB assay is capable of reliably detecting yeasts and Aspergillus spp. in clinical specimens and that the incorporation of both ITS1- and ITS2-targeted probes is required for optimal sensitivity. The test has potential utility in the early diagnosis of invasive fungal infection, since "fungal" DNA was detected in all 27 specimens. Prior to incorporation of probes to detect other fungal species, ITS sequencing may be performed to achieve species identification. %Z FOR Codes: 110899 110309 110899 110309 %0 Journal Article %~ PubMed %A Jones, Peter %A Turner, Kylie %A Djordjevic, Julianne %A Sorrell, Tania %A Wright, Lesley %A George, Anthony %T Role of Conserved Active Site Residues in Catalysis by Phospholipase B1 from Cryptococcus neoformans. %B Biochemistry %D 2007 %C United States %I American Chemical Society %V 46 %N 35 %P 10024-32 %@ 0006-2960 %X Phospholipase B1 (PLB1), secreted by the pathogenic yeast Cryptococcus neoformans, has an established role in virulence. Although the mechanism of its phospholipase B, lysophospholipase, and lysophospholipase transacylase activities is unknown, it possesses lipase, subtilisin protease aspartate, and phospholipase motifs containing putative catalytic residues S146, D392, and R108, respectively, conserved in fungal PLBs and essential for human cytosolic phospholipase A2 (cPLA2) catalysis. To determine the role of these residues in PLB1 catalysis, each was substituted with alanine, and the mutant cDNAs were expressed in Saccharomyces cerevisiae. The mutant PLB1s were deficient in all three enzymatic activities. As the active site structure of PLB1 is unknown, a homology model was developed, based on the X-ray structure of the cPLA2 catalytic domain. This shows that the two proteins share a closely related fold, with the three catalytic residues located in identical positions as part of a single active site, with S146 and D392 forming a catalytic dyad. The model suggests that PLB1 lacks the "lid" region which occludes the cPLA2 active site and provides a mechanism of interfacial activation. In silico substrate docking studies with cPLA2 reveal the binding mode of the lipid headgroup, confirming the catalytic dyad mechanism for the cleavage of the sn-2 ester bond within one of two separate binding tracts for the lipid acyl chains. Residues specific for binding arachidonic and palmitic acids, preferred substrates for cPLA2 and PLB1, respectively, are identified. These results provide an explanation for differences in substrate specificity between lipases sharing the cPLA2 catalytic domain fold and for the differential effect of inhibitors on PLB1 enzymatic activities. %Z FOR Codes: %0 Journal Article %~ PubMed %A Obando, Daniel %A Widmer, Fred %A Wright, Lesley C %A Sorrell, Tania C %A Jolliffe, Katrina A %T Synthesis, antifungal and antimicrobial activity of alkylphospholipids. %B Bioorganic & medicinal chemistry %D 2007 %C United Kingdom %I Pergamon %V 15 %N 15 %P 5158-5165 %@ 0968-0896 %X The antifungal, antibacterial and haemolytic activity of a series of alkylphosphocholines (e.g., miltefosine) and alkylglycerophosphocholines (e.g., edelfosine) has been investigated. These compound classes exhibit significant antifungal and moderate antibacterial activities. Several new alkylphosphocholine derivatives with amide or ester bonds in the alkyl chain have been synthesised. These compounds show much lower haemolytic activity than miltefosine. Alkylphosphocholines and alkylglycerophosphocholines show significant promise as novel orally available antifungal and antibacterial therapeutics. %Z FOR Codes: 111503 110899 %0 Journal Article %~ PubMed %A Ng, Clarissa K L %A Singhal, Vatsala %A Widmer, Fred %A Wright, Lesley C %A Sorrell, Tania C %A Jolliffe, Katrina A %T Synthesis, antifungal and haemolytic activity of a series of bis(pyridinium)alkanes. %B Bioorganic & medicinal chemistry %D 2007 %C United Kingdom. %I Pergamon %V 15 %N 10 %P 3422-3429 %@ 0968-0896 %X A series of bis(pyridinium)alkanes have been prepared and their antifungal activity, haemolytic activity and ability to inhibit fungal phospholipase B1 have been investigated, together with those of the commercially available antiseptics octenidine and dequalinium. Removal of the amino substituents from the pyridinium rings resulted in a significant decrease in antifungal activity. However, shortening or removing the alkyl chains attached to the amino groups had little effect on antifungal activity and significantly reduced haemolytic activity. Only octenidine was a strong inhibitor of fungal phospholipase B1. %Z FOR Codes: 111503 %0 Journal Article %~ PubMed %A Chen, Sharon %A Slavin, Monica %A Nguyen, Quoc %A Marriott, Deborah %A Playford, E Geoffrey %A Ellis, David %A Sorrell, Tania %A , Australian Candidemia Study %T Active surveillance for candidemia, Australia. %B Emerging infectious diseases %D 2006 %C United States %I US Department of Health and Human Services %V 12 %N 10 %P 1508-16 %@ 1080-6040 %X Population-based surveillance for candidemia in Australia from 2001 to 2004 identified 1,095 cases. Annual overall and hospital-specific incidences were 1.81/100,000 and 0.21/1,000 separations (completed admissions), respectively. Predisposing factors included malignancy (32.1%), indwelling vascular catheters (72.6%), use of antimicrobial agents (77%), and surgery (37.1%). Of 919 episodes, 81.5% were inpatient healthcare associated (IHCA), 11.6% were outpatient healthcare associated (OHCA), and 6.9% were community acquired (CA). Concomitant illnesses and risk factors were similar in IHCA and OHCA candidemia. IHCA candidemia was associated with sepsis at diagnosis (p<0.001), death <30 days after infection (p<0.001), and prolonged hospital admission (p<0.001). Non-Candida albicans species (52.7%) caused 60.5% of cases acquired outside hospitals and 49.9% of IHCA candidemia (p = 0.02). The 30-day death rate was 27.7% in those > or =65 years of age. Adult critical care stay, sepsis syndrome, and corticosteroid therapy were associated with the greatest risk for death. Systematic epidemiologic studies that use standardized definitions for IHCA, OHCA, and CA candidemia are indicated. %Z FOR Codes: %0 Journal Article %~ PubMed %A Playford, E Geoffrey %A Webster, Angela C %A Sorrell, Tania C %A Craig, Jonathan C %T Antifungal agents for preventing fungal infections in non-neutropenic critically ill and surgical patients: systematic review and meta-analysis of randomized clinical trials. %B The Journal of antimicrobial chemotherapy %D 2006 %C United Kingdom %I Oxford University Press %V 57 %N 4 %P 628-38 %@ 0305-7453 %X OBJECTIVES: This study aims to systematically identify and summarize the effects of antifungal prophylaxis in non-neutropenic critically ill adult patients on all-cause mortality and the incidence of invasive fungal infections. METHODS: Systematic review and meta-analysis of randomized controlled trials in all languages comparing the prophylactic use of any antifungal agent or regimen with placebo, no antifungal or another antifungal agent or regimen in non-neutropenic critically ill adult patients. We searched the Cochrane Central Register of Controlled Trials (CENTRAL, The Cochrane Library, Issue 3, 2005), MEDLINE (1966 to 2 September 2005) and EMBASE (1980 to week 36, 2005). We also hand-searched reference lists, abstracts of conference proceedings and scientific meetings (1998-2004) and contacted authors of included studies and pharmaceutical manufacturers. The primary outcomes assessed were all-cause mortality and proven invasive fungal infections. Two reviewers independently applied selection criteria, performed quality assessment and extracted data using an intention-to-treat approach. Data were synthesized using the random effects model and expressed as relative risk with 95% confidence intervals. RESULTS: Twelve unique trials (eight comparing fluconazole and four ketoconazole with no antifungal or a non-absorbable agent) involving 1606 randomized patients were included. For both outcomes of total mortality and invasive fungal infections, almost all trials of fluconazole and ketoconazole separately showed a non-significant risk reduction with prophylaxis. When combined, fluconazole/ketoconazole reduced total mortality by one-quarter (relative risk 0.76, 95% confidence interval 0.59-0.97) and invasive fungal infections by about one-half (relative risk 0.46, 95% confidence interval 0.31-0.68). No significant increase in the incidence of infection or colonization with the azole-resistant fungal pathogens Candida glabrata or Candida krusei was demonstrated, although the confidence intervals of the summary effect measures were wide. Adverse effects requiring treatment discontinuation were not more common amongst patients receiving prophylaxis. Results across all trials were homogeneous despite considerable heterogeneity in clinical and methodological characteristics. CONCLUSIONS: Prophylaxis with fluconazole or ketoconazole in critically ill patients reduces invasive fungal infections by one-half and total mortality by one-quarter. Although no significant increase in azole-resistant Candida species associated with prophylaxis was demonstrated, trials were not powered to exclude such an effect. In patients at increased risk of invasive fungal infections, antifungal prophylaxis with fluconazole should be considered. %Z FOR Codes: 110309 110310 110323 %0 Journal Article %~ PubMed %A Playford, E G %A Webster, A C %A Sorrell, T C %A Craig, J C %T Antifungal agents for preventing fungal infections in non-neutropenic critically ill patients. %B Cochrane database of systematic reviews (Online) %D 2006 %C United Kingdom %I Update Software %V 1 %N 1 %P CD004920 %@ 1469-493X %X BACKGROUND: Invasive fungal infections, important causes of morbidity and mortality in critically ill patients, may be preventable with the prophylactic administration of antifungal agents. OBJECTIVES: This study aims to systematically identify and summarize the effects of antifungal prophylaxis in non-neutropenic critically ill adult patients on all-cause mortality and the incidence of invasive fungal infections. SEARCH STRATEGY: We searched the Cochrane Central Register of Controlled Trials (CENTRAL), (The Cochrane Library, Issue 3, 2005), MEDLINE (1966 to 2 September 2005), and EMBASE (1980 to week 36, 2005). We also handsearched reference lists, abstracts of conference proceedings and scientific meetings (1998 to 2004), and contacted authors of included studies and pharmaceutical manufacturers. SELECTION CRITERIA: We included randomized controlled trials in all languages comparing the prophylactic use of any antifungal agent or regimen with placebo, no antifungal, or another antifungal agent or regimen in non-neutropenic critically ill adult patients. DATA COLLECTION AND ANALYSIS: Two authors independently applied selection criteria, performed quality assessment, and extracted data using an intention-to-treat approach. We resolved differences by discussion. We synthesized data using the random effects model and expressed results as relative risk with 95% confidence intervals. MAIN RESULTS: We included 12 unique trials (eight comparing fluconazole and four ketoconazole with no antifungal or a nonabsorbable agent) involving 1606 randomized patients. For both outcomes of total mortality and invasive fungal infections, almost all trials of fluconazole and ketoconazole separately showed a non-significant risk reduction with prophylaxis. When combined, fluconazole/ketoconazole reduced total mortality by about 25% (relative risk 0.76, 95% confidence interval 0.59 to 0.97) and invasive fungal infections by about 50% (relative risk 0.46, 95% confidence interval 0.31 to 0.68). We identified no significant increase in the incidence of infection or colonization with the azole-resistant fungal pathogens Candida glabrata or C. krusei, although the confidence intervals of the summary effect measures were wide. Adverse effects were not more common amongst patients receiving prophylaxis. Results across all trials were homogeneous despite considerable heterogeneity in clinical and methodological characteristics. AUTHORS'' CONCLUSIONS: Prophylaxis with fluconazole or ketoconazole in critically ill patients reduces invasive fungal infections by one half and total mortality by one quarter. Although no significant increase in azole-resistant Candida species associated with prophylaxis was demonstrated, trials were not powered to exclude such an effect. In patients at increased risk of invasive fungal infections, antifungal prophylaxis with fluconazole should be considered. %Z FOR Codes: 110399 %0 Journal Article %~ PubMed %A Coen, Muireann %A Bodkin, Jennifer %A Power, Damla %A Bubb, William A %A Himmelreich, Uwe %A Kuchel, Philip W %A Sorrell, Tania C %T Antifungal effects on metabolite profiles of medically important yeast species measured by nuclear magnetic resonance spectroscopy. %B Antimicrobial agents and chemotherapy %D 2006 %C United States %I American Society for Microbiology %V 50 %N 12 %P 4018-26 %@ 0066-4804 %X Drug-induced inhibition of fungal growth is used in the diagnostic laboratory to predict therapeutic efficacy but is relatively slow, and determination of endpoints can be problematic. Nuclear magnetic resonance (NMR) spectroscopy identifies the metabolic complement of microorganisms while monitoring utilization of constituents of the incubation medium. This technique may provide a rapid and objective indicator of antifungal effects. We evaluated the effects of caspofungin, amphotericin B (AMB), and voriconazole on metabolic profiles of yeast species cultured in RPMI-2% glucose-morpholinepropanesulfonic acid buffer in microtiter plates in a proof-of-principle study. 1H NMR spectra were obtained using Bruker NMR spectrometers at 1H frequencies of 600 and 360 MHz. Metabolites were identified by two-dimensional correlation NMR spectra, and relative peak integrals were calculated from one-dimensional 1H NMR spectra. MICs were determined by a modification of the Clinical and Laboratory Standards Institute broth microdilution method M27-A. Utilization of glucose and branched-chain and aromatic amino acid substrates was accompanied by fungal production of acetate, acetaldehyde, ethanol, formate, fumarate, glycerol, lactate, pyruvate, and succinate. Clear-cut metabolic endpoints indicating a greater than 50% reduction in substrate utilization and fungal metabolite production which correlated with MICs were noted at 16 and 24 h for all three drugs. At 8 h, reductions of greater than 50% for selected metabolites were noted for caspofungin and AMB. Direct NMR-based observation of metabolic alterations in yeast cultures reveals changes in key metabolic pathways and should be evaluated formally as a rapid technique for determining susceptibility to antifungal drugs. %Z FOR Codes: 110101 110801 %0 Journal Article %~ PubMed %A Sorrell, Tania C %A Wright, Lesley C %A Malik, Richard %A Himmelreich, Uwe %T Application of proton nuclear magnetic resonance spectroscopy to the study of Cryptococcus and cryptococcosis. %B FEMS yeast research %D 2006 %C United Kingdom %I Blackwell Publishing Ltd. %V 6 %N 4 %P 558-66 %@ 1567-1356 %X Proton nuclear magnetic resonance spectroscopy is a nondestructive technique that identifies chemicals in solution and in living cells. It has been used in cryptococcal research to identify the primary structure of capsular glucuronoxylomannans, link cellular apoptosis susceptibility (CAS) genes to positioning of residues on the mannose backbone of glucuronoxylomannan, and verify that the cryptococcal virulence determinant, phospholipase B, is elaborated in vivo. Promising clinical applications include speciation (Cryptococcus neoformans and Cryptococcus gattii), with preliminary evidence that varieties neoformans and grubii can also be distinguished, non-invasive diagnosis of cerebral cryptococcomas, and, in cases of meningitis, monitoring therapeutic response by analysis of cerebrospinal fluid. %Z FOR Codes: 110899 110309 %0 Journal Article %~ Isi %A Marriott, D. %A Playford, E. G. %A Nguyen, Q. %A Chen, S. %A Ellis, D. %A Slavin, M. %A Sorrell, T. %T Candidaemia in the Australian intensive care unit: Epidemiology, clinical features and outcome from a 3 year nationwide study. %B International Journal of Infectious Diseases %D 2006 %C United Kingdom %I Elsevier Ltd. %V 10 %N %P S77-S78 %@ 1201-9712 %X %Z FOR Codes: %0 Journal Article %~ Isi %A Anaissie, E. J. %A Segal, B. H. %A Graybill, J. R. %A Arndt, C. %A Perfect, J. R. %A Kleinberg, M. %A Pappas, P. %A Benjamin, D. %A Rubin, R. %A Aberg, J. A. %A Adderson, E. E. %A Adler-Shohet, F. C. %A Akan, H. %A Akova, M. %A Almyroudis, N. G. %A Alexander, B. D. %A Andes, D. %A Arrieta, A. %A Baddley, J. W. %A Barron, M. A. %A Belzberg, H. %A Boucher, H. W. %A Boyce, T. G. %A Casadevall, A. %A Chandrasekar, P. H. %A Cleary, J. D. %A Cordonnier, C. %A Cornely, O. A. %A Cuenca-Estrella, M. %A Daly, J. S. %A Daoura, N. %A Denning, D. W. %A dePauw, B. %A de Repentigny, L. %A Dignani, M. C. %A Dismukes, W. E. %A Donnelly, J. P. %A Donowitz, G. R. %A Dupont, B. %A Drusano, G. %A Ellis, M. %A Espinel-Ingroff, A. %A Fishman, J. A. %A Fleming, R. %A Forrest, G. %A Ghannoum, M. %A Goldman, M. %A Grazziutti, M. %A Greene, J. N. %A Greenberg, R. N. %A Gubbins, P. O. %A Hadley, S. %A Herbrecht, R. %A Hiemenz, J. W. %A Hope, W. %A Hospenthal, D. R. %A Husain, S. %A Ito, J. I. %A Jacobson, R. M. %A Johnson, M. %A Keating, M. R. %A Kett, D. H. %A Knapp, K. %A Kontoyiannis, D. P. %A Krcmery, V. C. %A Larsen, R. %A Laverdiere, M. %A Ljungman, P. %A Lortholary, O. %A Maertens, J. %A Marriott, D. %A Mattiuzzi, G. %A McGinnis, M. R. %A Morris, M. %A Nucci, M. %A Odds, F. C. %A Pankey, G. A. %A Patterson, T. %A Pfaller, M. %A Razonable, R. R. %A Reboli, A. C. %A Rinaldi, M. G. %A Roberts, G. D. %A Tudela, J. L. %A Rotstein, C. %A Ruhnke, M. %A Schuster, M. %A Shoham, S. %A Sia, I. G. %A Siebel, N. %A Silviera, F. %A Singh, N. %A Sobel, J. %A Solomkin, J. S. %A Sorrell, T. C. %A Steinbach, W. J. %A Temesgen, Z. %A Tortorano, A. %A Vartivarian, S. %A VerWeij, P. %A Viscoli, C. %A Viviani, M. A. %A Walker, R. C. %A Wheat, J. L. %A Wiley, J. %A Williamson, P. %A Wingard, J. R. %A Yu, V. L. %A Zaoutis, T. %T Clinical research in the lay press: Irresponsible journalism raises a huge dose of doubt. %B Clinical Infectious Diseases %D 2006 %C United States %I University of Chicago Press %V 43 %N 8 %P 1031-1039 %@ 1058-4838 %X %Z FOR Codes: %0 Journal Article %~ PubMed %A Ng, Clarissa K L %A Obando, Daniel %A Widmer, Fred %A Wright, Lesley C %A Sorrell, Tania C %A Jolliffe, Katrina A %T Correlation of antifungal activity with fungal phospholipase inhibition using a series of bisquaternary ammonium salts. %B Journal of medicinal chemistry %D 2006 %C United States %I Amer Chemical Soc %V 49 %N 2 %P 811-6 %@ 0022-2623 %X A series of bisquaternary ammonium salts with a 12-carbon spacer between the positive charges were synthesized, and their antifungal activity has been investigated. Compounds with butyl, pentyl, and isopentyl headgroups were the most potent antifungal agents with MICs in the range of 2.2-5.5 microM against both Cryptococcus neoformans and Candida albicans. The antifungal activity of these compounds correlated with their inhibition of cryptococcal phospholipase B1 (PLB1), a newly identified virulence factor. This indicates that the mode of action of these compounds may be inhibition of the fungal PLB1 enzyme, further validating this enzyme as a target for the development of novel antifungal therapies. %Z FOR Codes: %0 Journal Article %~ PubMed %A Wright, Lesley C %A Santangelo, Rosemary M %A Ganendren, Ranjini %A Payne, Jackie %A Djordjevic, Julianne T %A Sorrell, Tania C %T Cryptococcal lipid metabolism: phospholipase B1 is implicated in transcellular metabolism of macrophage-derived lipids. %B Eukaryotic cell %D 2006 %C USA %I American Society for Microbiology %V 6 %N 1 %P 37-47 %@ 1535-9778 %X Cryptococci survive and replicate within macrophages and can use exogenous arachidonic acid for the production of eicosanoids. Phospholipase B1 (PLB1) has a putative, but uninvestigated, role in these processes. We have shown that uptake and esterification of radiolabeled arachidonic, palmitic, and oleic acids by the Cryptococcus neoformans var. grubii H99 wild-type strain and its PLB1 deletion mutant strain (the Deltaplb1 strain) are independent of PLB1, except under hyperosmolar stress. Similarly, PLB1 was required for metabolism of 1-palmitoyl lysophosphatidylcholine (LysoPC), which is toxic to eukaryotic cell membranes, under hyperosmolar conditions. During both logarithmic and stationary phases of growth, the physiologically relevant phospholipids, dipalmitoyl phosphatidylcholine (DPPC) and dioleoyl phosphatidylcholine, were taken up and metabolized via PLB1. Exogenous DPPC did not enhance growth in the presence of glucose as a carbon source but could support it for at least 24 h in glucose-free medium. Detoxification of LysoPC by reacylation occurred in both the H99 wild-type and the Deltaplb1 strains in the presence of glucose, but PLB1 was required when LysoPC was the sole carbon source. This indicates that both energy-independent (via PLB1) and energy-dependent transacylation pathways are active in cryptococci. Phospholipase A(1) activity was identified by PLB1-independent degradation of 1-palmitoyl-2-arachidonoyl phosphatidylcholine, but the arachidonoyl LysoPC formed was not detoxified by reacylation. Using the human macrophage-like cell line THP-1, we demonstrated the PLB1-dependent incorporation of macrophage-derived arachidonic acid into cryptococcal lipids during cryptococcus-phagocyte interaction. This pool of arachidonate can be sequestered for eicosanoid production by the fungus and/or suppression of host phagocytic activity, thus diminishing the immune response. %Z FOR Codes: 110899 110104 %0 Journal Article %~ PubMed %A Lau, Anna %A Chen, Sharon %A Sorrell, Tania %A Carter, Dee %A Malik, Richard %A Martin, Patricia %A Halliday, Catriona %T Development and clinical application of a panfungal PCR assay to detect and identify fungal DNA in tissue specimens. %B Journal of clinical microbiology %D 2006 %C United States %I American Society for Microbiology %V 45 %N %P 380-5 %@ 0095-1137 %X Given the rise in the incidence of invasive fungal infections (IFIs) and the expanding spectrum of fungal pathogens, early and accurate identification of the causative pathogen is essential. We developed a panfungal PCR assay that targets the internal transcribed spacer 1 (ITS1) region of the ribosomal DNA gene cluster to detect fungal DNA in fresh and formalin-fixed, paraffin-embedded (PE) tissue specimens from patients with culture-proven (n=38) or solely histologically proven (n=24) IFIs. PCR products were sequenced and compared with sequences in the GenBank database to identify the causal pathogen. The molecular identification was correlated with results from histological examination and culture. The assay successfully detected and identified the fungal pathogen in 93.6% and 64.3% of culture-proven and solely histologically proven cases of IFI, respectively. A diverse range of fungal genera were identified, including species of Candida, Cryptococcus, Trichosporon, Aspergillus, Fusarium, Scedosporium, Exophiala, Exserohilum, Apophysomyces, Actinomucor, and Rhizopus. For five specimens, molecular analysis identified a pathogen closely related to that identified by culture. All PCR-negative specimens (n=10) were PE tissues in which fungal hyphae were visualized. The results support the use of the panfungal PCR assay in combination with conventional laboratory tests for accurate identification of fungi in tissue specimens. %Z FOR Codes: 110899 %0 Journal Article %~ PubMed %A Widmer, Fred %A Wright, Lesley C %A Obando, Daniel %A Handke, Rosemary %A Ganendren, Ranjini %A Ellis, David H %A Sorrell, Tania C %T Hexadecylphosphocholine (miltefosine) has broad-spectrum fungicidal activity and is efficacious in a mouse model of cryptococcosis. %B Antimicrobial agents and chemotherapy %D 2006 %C United States %I American Society for Microbiology %V 50 %N 2 %P 414-21 %@ 0066-4804 %X The alkyl phosphocholine drug miltefosine is structurally similar to natural substrates of the fungal virulence determinant phospholipase B1 (PLB1), which is a potential drug target. We determined the MICs of miltefosine against key fungal pathogens, correlated antifungal activity with inhibition of the PLB1 activities (PLB, lysophospholipase [LPL], and lysophospholipase-transacylase [LPTA]), and investigated its efficacy in a mouse model of disseminated cryptococcosis. Miltefosine inhibited secreted cryptococcal LPTA activity by 35% at the subhemolytic concentration of 25 microM (10.2 microg/ml) and was inactive against mammalian pancreatic phospholipase A2 (PLA2). At 250 microM, cytosolic PLB, LPL, and LPTA activities were inhibited by 25%, 51%, and 77%, respectively. The MICs at which 90% of isolates were inhibited (MIC90s) against Candida albicans, Candida glabrata, Candida krusei, Cryptococcus neoformans, Cryptococcus gattii, Aspergillus fumigatus, Fusarium solani, Scedosporium prolificans, and Scedosporium apiospermum were 2 to 4 microg/ml. The MICs of miltefosine against Candida tropicalis (n = 8) were 2 to 4 microg/ml, those against Aspergillus terreus and Candida parapsilosis were 8 microg/ml (MIC90), and those against Aspergillus flavus (n = 8) were 2 to 16 microg/ml. Miltefosine was fungicidal for C. neoformans, with rates of killing of 2 log units within 4 h at 7.0 microM (2.8 microg/ml). Miltefosine given orally to mice on days 1 to 5 after intravenous infection with C. neoformans delayed the development of illness and mortality and significantly reduced the brain cryptococcal burden. We conclude that miltefosine has broad-spectrum antifungal activity and is active in vivo in a mouse model of disseminated cryptococcosis. The relatively small inhibitory effect on PLB1 enzyme activities at concentrations exceeding the MIC by 2 to 20 times suggests that PLB1 inhibition is not the only mechanism of the antifungal effect. %Z FOR Codes: 110309 110309 110899 %0 Journal Article %~ PubMed %A Siafakas, A Rosemary %A Wright, Lesley C %A Sorrell, Tania C %A Djordjevic, Julianne T %T Lipid rafts in Cryptococcus neoformans concentrate the virulence determinants phospholipase B1 and Cu/Zn superoxide dismutase. %B Eukaryotic cell %D 2006 %C United States %I American Society for Microbiology %V 5 %N 3 %P 488-98 %@ 1535-9778 %X Lipid rafts have been identified in the membranes of mammalian cells, the yeast Saccharomyces cerevisiae, and the pathogenic fungus Candida albicans. Formed by a lateral association of sphingolipids and sterols, rafts concentrate proteins carrying a glycosylphosphatidylinositol (GPI) anchor. We report the isolation of membranes with the characteristics of rafts from the fungal pathogen Cryptococcus neoformans. These characteristics include insolubility in Triton X-100 (TX100) at 4 degrees C, more-buoyant density within a sucrose gradient than the remaining membranes, and threefold enrichment with sterols. The virulence determinant phospholipase B1 (PLB1), a GPI-anchored protein, was highly concentrated in raft membranes and could be displaced from them by treatment with the sterol-sequestering agent methyl-beta-cyclodextrin (MbetaCD). Phospholipase B enzyme activity was inhibited in the raft environment and increased 15-fold following disruption of rafts with TX100 at 37 degrees C. Treatment of viable cryptococcal cells in suspension with MbetaCD also released PLB1 protein and enzyme activity, consistent with localization of PLB1 in plasma membrane rafts prior to secretion. The antioxidant virulence factor Cu/Zn superoxide dismutase (SOD1) was concentrated six- to ninefold in raft membrane fractions compared with nonraft membranes, whereas the cell wall-associated virulence factor laccase was not detected in membranes. We hypothesize that raft membranes function to cluster certain virulence factors at the cell surface to allow efficient access to enzyme substrate and/or to provide rapid release to the external environment. %Z FOR Codes: 110899 110104 %0 Journal Article %~ PubMed %A Turner, Kylie M %A Wright, Lesley C %A Sorrell, Tania C %A Djordjevic, Julianne T %T N-linked glycosylation sites affect secretion of cryptococcal phospholipase B1, irrespective of glycosylphosphatidylinositol anchoring. %B Biochimica et biophysica acta %D 2006 %C Netherlands %I Elsevier BV %V 1760 %N %P 1569-79 %@ 0006-3002 %X Secreted phospholipase B enzymes (PLB1) with high levels of N-linked glycosylation are proven fungal virulence determinants. We demonstrated that removal of N-linked glycans from secreted cryptococcal PLB1 leads to loss of enzyme activity. To determine if individual N-glycan attachment sites affect secretion of active enzyme, we altered three along the entire length of the protein, by site-directed mutagenesis, namely Asn56, Asn430 and Asn550 to Ala, in wild-type PLB1 (full length) and a glycosylphosphatidylinositol (GPI) anchorless version (PLB1(GPI-)) that is hypersecreted due to lack of membrane association. Alteration of Asn56 and Asn550 in both PLB1 and PLB1(GPI-) abolished enzyme secretion while alteration of Asn430 reduced secretion by 60%, following expression in Saccharomyces cerevisiae. Reduced secretion coincided with reduced enzyme in membranes and cell walls confirming a reduction in the rate of PLB1 transport to the cell surface. Deglycosylation of cryptococcal PLB1 increased its susceptibility to proteolysis suggesting that the absence of full glycosylation status leads to degradation of unstable PLB1, resulting in reduced traffic through the secretory pathway. We conclude that individual N-linked glycans are required for optimal transport of PLB1 to the cell surface and optimal secretion of both PLB1 and PLB1(GPI-). %Z FOR Codes: 110899 %0 Journal Article %~ PubMed %A Ganendren, Ranjini %A Carter, Elizabeth %A Sorrell, Tania %A Widmer, Fred %A Wright, Lesley %T Phospholipase B activity enhances adhesion of Cryptococcus neoformans to a human lung epithelial cell line. %B Microbes and infection / Institut Pasteur %D 2006 %C France %I Elsevier France %V 8 %N 4 %P 1006-15 %@ 1286-4579 %X Secreted phospholipase B (PLB1), which contains three enzyme activities in the one protein, is necessary for the initiation of pulmonary infection by Cryptococcus neoformans and for dissemination from the lung via the lymphatics and blood. Adhesion to lung epithelium is the first step in this process, therefore we investigated the role of PLB1 in adhesion to a human lung epithelial cell line, A549, using C. neoformans var. grubii wild-type strain H99, a PLB1 deletion mutant (deltaplb1), and a reconstituted strain (deltaplb1rec). Adhesion of H99 and deltaplb1rec was approximately 69% greater than deltaplb1 at 4 h. Adhesion of deltaplb1 significantly increased after killing by chemicals or heat, and Fourier-transformed analysis by FTIR spectroscopy indicated this was due to changes in capsular and/or cell wall polysaccharides and proteins. Inhibition by specific PLB1 antibodies, or inhibitors of phospholipase B (PLB), but not lysophospholipase (LPL) or lysophospholipase transacylase (LPTA) activities decreased the adhesion of H99 and deltaplb1rec by 33-58%. Growth under conditions of osmotic stress and high glucose concentration increased both PLB secretion and subsequent cryptococcal adhesion. Dose-dependent increases (to 67%) in adhesion of live deltaplb1 were observed in the presence of 0.1-2 mM palmitic acid. We conclude that PLB1 plays a role in the binding of C. neoformans to host lung epithelial cells, possibly due to production of fatty acids from plasma membranes and/or surfactant by PLB activity. %Z FOR Codes: 110104 110899 %0 Journal Article %~ PubMed %A Halliday, Catriona %A Hoile, Rebecca %A Sorrell, Tania %A James, Greg %A Yadav, Satya %A Shaw, Peter %A Bleakley, Marie %A Bradstock, Ken %A Chen, Sharon %T Role of prospective screening of blood for invasive aspergillosis by polymerase chain reaction in febrile neutropenic recipients of haematopoietic stem cell transplants and patients with acute leukaemia. %B British journal of haematology %D 2006 %C United Kingdom %I Blackwell Publishing Ltd. %V 132 %N 4 %P 478-86 %@ 0007-1048 %X Guidelines for the use of polymerase chain reaction (PCR)-based assays to aid the diagnosis of invasive aspergillosis (IA) in high-risk haematology patients have not been formulated. We prospectively evaluated a nested PCR assay to detect Aspergillus in blood during 95 febrile neutropenic episodes, in patients with haematological malignancy and haematopoietic stem cell transplant (HSCT) recipients. PCR results were correlated with the diagnostic classification of the 2002 European Organisation for Research and Treatment of Cancer/Mycosis Study Group. When two-positive results were used to define an episode as ''PCR positive'', the sensitivity, specificity, positive-predictive value and negative predictive value for ''proven''/''probable'' IA (n = 13) were 100%, 75.4%, 46.4% and 100%, respectively. Consecutive positive results occurred in 61.5% of these 13 episodes. Overall, PCR positivity preceded standard diagnosis by a mean of 14 d and the median time between positive results was shorter than that in other categories of IA. All 13 episodes occurred in the setting of allogeneic HSCT recipients and acute leukaemia. If ''eligibility'' for antifungal therapy were based on two-positive-PCR tests, use of empiric treatment could have been reduced by up to 37%. The nested PCR assay is a practical screening test for excluding IA. Patients with consecutive positive results or intermittent-positive results (within 14 d) warrant immediate investigations for IA and the initiation of antifungal therapy. %Z FOR Codes: 110899 110202 110309 %0 Journal Article %~ PubMed %A Playford, E Geoffrey %A Kong, Fanrong %A Sun, Ying %A Wang, Hui %A Halliday, Catriona %A Sorrell, Tania C %T Simultaneous detection and identification of Candida, Aspergillus, and Cryptococcus species by reverse line blot hybridization. %B Journal of clinical microbiology %D 2006 %C United States %I American Society for Microbiology %V 44 %N 3 %P 876-80 %@ 0095-1137 %X We report on a reverse line blot (RLB) assay, utilizing fungal species-specific oligonucleotide probes to hybridize with internal transcribed spacer 2 region sequences amplified using a nested panfungal PCR. Reference and clinical strains of 16 Candida species (116 strains), Cryptococcus neoformans (five strains of Cryptococcus neoformans var. neoformans, five strains of Cryptococcus neoformans var. grubii, and six strains of Cryptococcus gatti), and five Aspergillus species (68 strains) were all correctly identified by the RLB assay. Additional fungal species (16 species and 26 strains) not represented on the assay did not exhibit cross-hybridization with the oligonucleotide probes. In simulated clinical specimens, the sensitivity of the assay for Candida spp. and Aspergillus spp. was 10(0.5) cells/ml and 10(2) conidia/ml, respectively. This assay allows sensitive and specific simultaneous detection and identification of a broad range of fungal pathogens. %Z FOR Codes: 110899 %0 Journal Article %~ Isi %A Playford, E. G. %A Webster, A. C. %A Sorrell, T. C. %A Craig, J. C. %T Systematic review and meta-analysis of antifungal agents for preventing fungal infections in liver transplant recipients. %B European Journal of Clinical Microbiology & Infectious Diseases %D 2006 %C Germany %I Springer-Verlag %V 25 %N 9 %P 549-561 %@ 0934-9723 %X %Z FOR Codes: %0 Journal Article %~ PubMed %A Sorrell, Tania %A Lonsdale, Carey %T Urgent strategic research into influenza to inform health policy and protect the public. %B The Medical journal of Australia %D 2006 %C Australia %I Australasian Medical Publishing Company Pty. Ltd. %V 185 %N 10 %P S77-S79 %@ 0025-729X %X The Australian management plan for pandemic influenza (2005) highlighted a number of areas where more information may yield better plans for protecting Australia. In 2005, the National Health and Medical Research Council (NHMRC) developed a special "urgent research" funding program to meet those information needs as quickly as possible. The funding program resulted in grants totalling $6.5 million being awarded for 33 research projects, in five broad areas: Detection and identification of the virus; Vaccine development and evaluation; Antiviral medication use and effectiveness; Public health interventions; and Understanding behavioural responses to achieve effective communication and staged implementation of public health strategies. Outcomes of the program will be evaluated formally in 2007. %Z FOR Codes: 110804 111702