The synergistic effect of minocycline and azole antifungal drugs against Scedosporium and Lomentospora species.

BACKGROUND: This study was aimed to determine the potency of Minocycline (MIN) and azoles, including itraconazole (ITR), voriconazole (VOR) and posaconazole (POS) against Scedosporium and Lomentospora species. RESULTS: This study revealed that MIN exhibited no significant antifungal activity against any of the tested strains, whereas in vitro combination of MIN with ITR, VOR or POS showed satisfactory synergistic effects against 8 (80%), 1 (10%), and 9 (90%) strains, respectively. Moreover, combined use of MIN with azoles decreased the minimum inhibitory concentration (MIC) range from 5.33-16 µg/ml to 1-16 µg/ml for ITR, from 0.42-16 µg/ml to 0.21-16 µg/ml for VOR, and from 1.33-16 µg/ml to 0.33-16 µg/ml for POS. Meanwhile, no antagonistic interactions were observed between the above combinations. The G. mellonella infection model demonstrated the in vivo synergistic antifungal effect of MIN and azoles. CONCLUSIONS: The present study demonstrated that combinations between MIN and azoles lead to synergistic antimicrobial effects on Scedosporium and Lomentospora species, while showing a potential for overcoming and preventing azole resistance.

A label-free electrochemical biosensor based on magnetic biocomposites with DNAzyme and hybridization chain reaction dual signal amplification for the determination of Pb2.

A highly sensitive and selective electrochemical biosensor for Pb2+ with a dual-amplification strategy is proposed. The first amplification step was realized by the cycle of Pb2+ and 8-17 DNAzyme (S2), and the hybridization chain reaction (HCR) triggered by S1 further amplified the electrochemical signal. Fe3O4@Au NPs, as a multifunctional magnetic carrier, is not only manifested in the construction of a magnetically controlled electrochemical response interface, but also has significant contribution in the purifying system, reducing interference, increasing the specific surface area, and the DNA loading. The magnetic nanocomposites were characterized by TEM as spheres with particle size of around 39 nm. When there was no Pb2+, long double-strand DNA (dsDNA) is formed on the surface of Fe3O4@Au NPs by the S1-triggered HCR; in the presence of Pb2+, S2 is activated and S1 on the surface of magnetic biocomposites (Fe3O4@Au NPs-S1) is continuously cleaved with the cycle of Pb2+ and S2, leading to a significant decrease of methylene blue (MB) absorbed on dsDNA. Such reverse dual-signal amplification strategy effectively increased the current difference and improved the sensitivity of the proposed sensor. The electrochemical signal of MB was obtained by differential pulse voltammetry (DPV) with preconcentration, showing a linear response toward Pb2+ ranging from 50 pM to 1 µM with a detection limit of 15 pM. The proposed method has feasible applications in detecting other heavy metal ions based on other metal-dependent DNAzyme. Graphical Abstract.

A label-free electrochemical magnetic aptasensor based on exonuclease III-assisted signal amplification for determination of carcinoembryonic antigen.

A novel label-free and exonuclease III (Exo III)-assisted signal amplification electrochemical aptasensor was constructed for the determination of carcinoembryonic antigen (CEA) via magnetic field-induced self-assembly of magnetic biocomposites (Fe3O4@Au NPs-S1-S2-S3). The magnetic biocomposites were acquired by modifying double-stranded DNA (S1-S2-S3) on the surface of Fe3O4@Au nanoparticles (Fe3O4@Au NPs). Among them, Fe3O4@Au NPs were used as carriers for magnetic separation, thiolated single-stranded DNA (S1) provided signal sequence, CEA aptamer (S2) worked as a recognition element, and complementary strand (S3) was used to form double strands. In the presence of CEA, S2 bonded with CEA competitively; the exposed S1 could not be cleaved since Exo III was inactive against ssDNA. The G-quadruplex/hemin complexes finally formed with the existence of K+, and the high electrochemical signal of G-quadruplex/hemin complexes was recorded by differential pulse voltammetry (DPV) at - 0.6 V. Conversely, in the absence of CEA, dsDNA was cleaved from the 3' blunt end by Exo III; the disappearance of G-rich sequence blocked the generation of the signal. This method exhibited good selectivity and sensitivity for the determination of CEA; the linear range was from 0.1 to 200 ng mL-1 and the limit of detection was 0.4 pg mL-1. Graphical abstract.

A signal transduction approach for multiplexed detection of transcription factors by integrating DNA nanotechnology, multi-channeled isothermal amplification, and chromatography.

The variation patterns of transcription factors (TFs) provide direct information for the states of cell populations, which is of significance for biomedical research and clinical diagnostics. Herein, we show that through multi-channeled isothermal amplification, it is feasible to connect DNA-based signal transduction with chromatography for multiplexed detection of TFs. The described system is referred to as "PAC" which includes three major steps: (i) Protection, which uses DNA-modified magnetic beads to capture TFs and converts the capturing event into triggering signal; (ii) Amplification, which receives the triggering signal and generate DNA reporters through multi-channeled extension and nicking of oligonucleotides; and (iii) Chromatography, which separates and detects the DNA reporters in liquid chromatography. The quantitative detection of five essential TFs includes p50, p53, AP-1, MITF, and c-Myc is realized in a multiplexed manner, with the lowest detection limit of 0.5 pM. PAC can also provide effective means to measure the above five TFs in real samples, including cultured cells, xenograft tumors, and blood-based liquid biopsy. This study not only established a solution for multiplexed measurement of TFs for molecular diagnostics, but also paved avenue for bridging the gap between DNA nanotechnology and chromatography.

Dual-Emission Reverse Change Ratio Photoluminescence Sensor Based on a Probe of Nitrogen-Doped Ti3C2 Quantum Dots@DAP to Detect H2O2 and Xanthine.

Titanium carbide quantum dots (Ti3C2 QDs) derived from two-dimensional (2D) Ti3C2Tx (MXene) are the rising-star material recently. Herein, nitrogen-doped Ti3C2 QDs (N-Ti3C2 QDs) were synthesized via a solvothermal method. The obtained N-Ti3C2 QDs exhibited excitation-dependent photoluminescence, antiphotobleaching, and dispersion stability. Furthermore, by combining the N-Ti3C2 QDs and DAP (2,3-diaminophenazine, the oxidative product of o-phenylenediamine) as a composite nanoprobe (N-Ti3C2 QDs@DAP), we developed a dual-emission reverse change ratiometric sensor to quantitatively monitor H2O2 based on photoinduced electron-transfer effects, where N-Ti3C2 QDs acted as the donor and DAP as the acceptor. On the basis of the xanthine converting into H2O2 through the catalysis of xanthine oxidase, the N-Ti3C2 QDs@DAP nanoprobe was also exploited for xanthine sensing. As a result, the proposed assay was demonstrated to be highly sensitive for H2O2 and xanthine with detection limits of 0.57 and 0.34 µM, respectively. In a word, we have investigated the application of N-Ti3C2 QDs in H2O2 and xanthine sensing and opened a new and exciting avenue for the N-Ti3C2 QDs in biosensing.

A novel tool to provide predictable alignment data irrespective of source and image quality acquired on mobile phones: what engineers can offer clinicians.

PURPOSE: Existing automated spine alignment is based on original X-rays that are not applicable for teleradiology for spinal deformities patients. We aim to provide a novel automated vertebral segmentation method enabling accurate sagittal alignment detection, with no restrictions imposed by image quality or pathology type. METHODS: A total of 428 optical images of original sagittal X-rays taken by smartphones or screenshots for consecutive patients attending our spine clinic were prospectively collected. Of these, 300 were randomly selected and their vertebrae were labelled with Labelme. The ground truth was specialists measured sagittal alignment parameters. Pre-trained Mask R-CNN was fine-tuned and trained to predict the vertebra level(s) on the remaining 128 testing cases. The sagittal alignment parameters including the thoracic kyphosis (TK), lumbar lordosis (LL) and sacral slope (SS) were auto-detected, based on the segmented vertebra. Dice similarity coefficient (DSC) and mean intersection over union (mIoU) were calculated to evaluate the accuracy of the predicted vertebra. The detected sagittal alignments were then quantitatively compared with the ground truth. RESULTS: The DSC was 84.6 ± 3.8% and mIoU was 72.1 ± 4.8% indicating accurate vertebra prediction. The sagittal alignments detected were all strongly correlated with the ground truth (p < 0.001). Standard errors of the estimated parameters had a small difference from the specialists' results (3.5° for TK and SS; 3.4° for LL). CONCLUSION: This is the first study using fine-tuned Mask R-CNN to predict vertebral locations on optical images of X-rays accurately and automatically. We provide a novel alignment detection method that has a significant application on teleradiology aiding out-of-hospital consultations. These slides can be retrieved under Electronic Supplementary Material.

Label-Free Colorimetric Detection of Acid Phosphatase and Screening of Its Inhibitors Based on Biomimetic Oxidase Activity of MnO2 Nanosheets.

In this research, we attempted to develop a sensitive colorimetric sensing strategy for the detection of acid phosphatase (ACP) based on MnO2 nanosheets and explored its applications in screening and evaluating inhibitors of ACP. The MnO2 nanosheets exhibit intrinsic biomimetic oxidase activity, which can catalyze the oxidation of the colorless 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonate) diammonium salt (ABTS) into green oxidized ABTS (oxABTS). Upon the introduction of ACP, l-ascorbic acid-2-phosphate can be dephosphorylated to ascorbic acid, which arouses the disintegration of MnO2 nanosheets into Mn2+ ions. This disintegration weakens the enzyme mimicking activity of the MnO2 nanosheets, leading to the impediment of the oxidation of ABTS. Conversely, in the absence of ACP, the ABTS is rapidly oxidized by MnO2, leading to a significant colorimetric signal change. The absorbance difference at 420 nm displayed a linear relationship with the concentration of ACP ranging from 0.075 to 0.45 mU·mL-1, generating a detection limit of 0.046 mU·mL-1. In the inhibition assays, this sensing platform provided simple detection for parathion-methyl (PM), a representative inhibitor of ACP. The facile evaluation of the inhibitory effect of PM, including its IC50 toward ACP, was also realized.

A versatile fluorometric aptasensing scheme based on the use of a hybrid material composed of polypyrrole nanoparticles and DNA-silver nanoclusters: application to the determination of adenosine, thrombin, or interferon-gamma.

The authors describe a versatile aptasensing scheme based on the use of polypyrrole nanoparticles (PPyNPs) and DNA-silver nanoclusters (DNA-AgNCs) for multiple target detection. The DNA-AgNCs consist of two functional domains, viz. (a) a nucleation domain for attaching the metal core of the nanoclusters, and (b) a recognition domain which consists of a single-stranded aptamer. In the absence of analytes, the single-strand recognition domain will be absorbed onto the surface of the PPyNPs through π stacking and hydrophobic interactions. As a result, the red fluorescence of the DNA-AgNCs (with excitation/emission peaks at 535/625 nm) is quenched by the PPyNPs. On introducing the analytes, the DNA-AgNCs will bind them. This leads to the desorption of DNA-AgNCs and the recovery of the red fluorescence. Based on the above strategy, a versatile, sensitive and selective aptasensor was established for detection of adenosine, thrombin and interferon-gamma. The method was applied to the detection of the above targets in (spiked) serum samples and gave satisfactory results, with detection limit of 0.58 nM for IFN-γ, 0.39 nM for adenosine, and 2.2 nM for thrombin. The use of PPyNPs results in uniquely low non-specific absorption and in improved analytical results in case of real-sample analysis when compared to previously reported methods. Graphical abstract Schematic illustration of DNA-silver nanoclusters and polypyrrole nanoparticles in an aptasensor for detection of multiple targets.

Detecting transcription factors with allosteric DNA-Silver nanocluster switches.

Sensitive and efficient detection of protein markers, such as transcription factors (TFs), is an important issue in postgenomic era. In this paper, we report a DNA nanodevice, allosteric DNA-silver nanocluster switches (AgSwitches), for TFs detection. The mechanism of this nanodevice is based on the binding-induced allostery whereby the binding between AgSwitches and TFs alters the conformation of AgSwitches. This alteration brings DNA-silver nanocluster (DNA-AgNCs) and guanine-rich enhancer sequences (GRS) into close proximity, generating fluorescent enhancement for quantifications. Our results revealed that the sequence design of AgSwitches can be rationally optimized according to stimulated free energy, and we demonstrated that this method can not only be used for detecting TFs in nuclear extracts of cells, but also be developed as a tool for screening inhibitors of TFs. Overall, this work expanded the category allosteric DNA nanodevices by first introducing DNA-AgNCs into this area, and the obtained method was efficient for TFs-related investigations.

Effects of Photodynamic Therapy on the Growth and Antifungal Susceptibility of Scedosporium and Lomentospora spp.

Scedosporium and Lomentospora species are the second most frequent colonizing, allergenic, or invasive fungal pathogens in patients with cystic fibrosis, and are responsible for infections varying from cutaneous and subcutaneous tissue infections caused by traumatic inoculation to severe systemic diseases in immunocompromised patients. The clinical relevance of fungal airway colonization for individual patients harboring Scedosporium and Lomentospora species is still an underestimated issue. The high resistance of Scedosporium and Lomentospora species to antifungal drugs has highlighted the need for alternative treatment modalities, and antimicrobial photodynamic therapy may be one such alternative. In this study, methylene blue was applied as a photosensitizing agent to 6 type strains of Scedosporium and Lomentospora species, and we irradiated the strains using a light-emitting diode (635 ± 10 nm, 12 J/cm2). We evaluated the effects of photodynamic therapy on strain growth and on the in vitro susceptibility of the strains to itraconazole, voriconazole, posaconazole, and amphotericin B. A colony-forming unit reduction of up to 5.2 log10 was achieved. Minimal inhibitory concentration ranges also decreased significantly with photoinactivation. Photodynamic therapy improved both the inactivation rates and the antifungal susceptibility profile of all fungal isolates tested.

INK128 Exhibits Synergy with Azoles against Exophiala spp. and Fusarium spp.

Infections of Exophiala spp. and Fusarium spp. are often chronic and recalcitrant. Systemic disseminations, which mostly occur in immunocompromised patients, are often refractory to available antifungal therapies. The conserved target of rapamycin (TOR) orchestrates cell growth and proliferation in response to nutrients and growth factors, which are important for pathogenicity and virulence. INK128 is a second-generation ATP-competitive TOR inhibitor, which binds the TOR catalytic domain and selectively inhibits TOR. In the present study, we investigated the in vitro activities of INK128 alone and the interactions of INK128 with conventional antifungal drugs including itraconazole, voriconazole, posaconazole, and amphotericin B against 18 strains of Exophiala spp. and 10 strains of Fusarium spp. via broth microdilution checkerboard technique system adapted from Clinical and Laboratory Standards Institute broth microdilution method M38-A2. INK128 alone was inactive against all isolates tested. However, favorable synergistic effects between INK128 and voriconazole were observed in 61% Exophiala strains and 60% Fusarium strains, despite Fusarium strains exhibited high MIC values (4-8 µg/ml) against voriconazole. In addition, synergistic effects of INK128/itraconazole were shown in 33% Exophiala strains and 30% Fusarium strains, while synergy of INK128/posaconazole were observed in 28% Exophiala strains and 30% Fusarium strains. The effective working ranges of INK128 were 0.125-2 µg/ml and 1-4 µg/ml against Exophiala isolates and Fusarium isolates, respectively. No synergistic effect was observed when INK128 was combined with amphotericin B. No antagonism was observed in all combinations. In conclusion, INK128 could enhance the in vitro antifungal activity of voriconazole, itraconazole and posaconazole against Exophiala spp. and Fusarium spp., suggesting that azoles, especially voriconazole, combined with TOR kinase inhibitor might provide a potential strategy to the treatment of Exophiala and Fusarium infections. However, further investigations are warranted to elucidate the underlying mechanism and to determine possible reliable and safe application in clinical practice.

Molecular identification and susceptibility of clinically relevant Scedosporium spp. in China.

As various new sibling species within the Scedosporium spp. have been described recently, this study was conducted to investigate distribution and antifungal susceptibility profiles of the different species of Scedosporium spp. in China. Twenty-one clinical strains of Scedosporium from China and two strains from Japan were reidentified by MLSA. The analysis included BT2, CAL, RPB, SOD, and ACT and the combination of the five loci. Pseudallescheria boydii complex (17 strains) and S. apiospermum (6 strains) were identified. P. boydii complex included four closely related subgroups: P. boydii (9 strains), P. ellipsoidea (6 strains), P. fusoidea (1 strain), and P. angusta (1 strain). There were no significant differences in MICs for neither VOR, POS, nor AMB over all the five species in study. For itraconazole, intraspecific diversity was evident.

Cognitive behavioral therapy in combination with systemic family therapy improves mild to moderate postpartum depression.

OBJECTIVE: To explore the effect of cognitive behavioral therapy (CBT) in combination with systemic family therapy (SFT) on mild to moderate postpartum depression and sleep quality. METHODS: 249 primiparous women with mild to moderate postpartum depression were recruited and randomly assigned to a control group (n=128), which received conventional postpartum care, or to a psychological intervention group (n=121), which received conventional postpartum care combined with psychological intervention. The Edinburgh Postnatal Depression Scale (EPDS) and Pittsburgh Sleep Quality Index (PSQI) were employed to evaluate depression and sleep quality, respectively. RESULTS: 104 patients in the intervention group and 109 in the control group completed the study. After intervention, the EPDS score, PSQI score, sleep quality score, sleep latency score, sleep duration score, habitual sleep efficiency score, sleep disturbance score, and daytime dysfunction score were significantly lower in the intervention group than in the control group. The EPDS and PSQI scores of each group at different time points after intervention were markedly decreased compared with those before intervention, and the reduction in the intervention group was more evident than that in the control group. CONCLUSION: CBT in combination with SFT can improve depression and sleep quality in patients with mild to moderate postpartum depression.

Cognitive behavioral therapy in combination with systemic family therapy improves mild to moderate postpartum depression

Objective: To explore the effect of cognitive behavioral therapy (CBT) in combination with systemic family therapy (SFT) on mild to moderate postpartum depression and sleep quality. Methods: 249 primiparous women with mild to moderate postpartum depression were recruited and randomly assigned to a control group (n=128), which received conventional postpartum care, or to a psychological intervention group (n=121), which received conventional postpartum care combined with psychological intervention. The Edinburgh Postnatal Depression Scale (EPDS) and Pittsburgh Sleep Quality Index (PSQI) were employed to evaluate depression and sleep quality, respectively. Results: 104 patients in the intervention group and 109 in the control group completed the study. After intervention, the EPDS score, PSQI score, sleep quality score, sleep latency score, sleep duration score, habitual sleep efficiency score, sleep disturbance score, and daytime dysfunction score were significantly lower in the intervention group than in the control group. The EPDS and PSQI scores of each group at different time points after intervention were markedly decreased compared with those before intervention, and the reduction in the intervention group was more evident than that in the control group. Conclusion: CBT in combination with SFT can improve depression and sleep quality in patients with mild to moderate postpartum depression. .

[Quantitative real-time polymerase chain reaction for the diagnosis of invasive fungal disease].

OBJECTIVE: To establish the diagnostic assays of quantitative real-time polymerase chain reaction (PCR) for the diagnosis of invasive fungal disease and evaluate their accuracy in clinical samples. METHODS: Three assays have been developed for the diagnosis of clinically prevalent fungi, Aspergillus spp. and Candida spp.. Their analytical sensitivity and specificity were evaluated. Twenty-one serum samples from invasive fungal disease patients and non-invasive fungal disease patients were analyzed by the diagnostic assays and the accuracy was evaluated. RESULTS: Three assays were managed to run at the same condition. The universal fungi assay was able to detect, but not differentiate between most of the pathogenic fungi, including A. fumigatus, A. flavus, A. niger, C. albicans, C. parapsilosis, C. tropicalis, C.kruseii, C.glabrata, Cryptococcus neoformans, Rhizomucor variabilis, Rhizopus arrhizus and Penicillosis marneffei. The assays of Aspergillus spp. and Candida spp. were able to detect, but not differentiate between A. fumigatus, A. flavus, A. niger and C. albicans, C. parapsilosis, C. tropicalis. The detection limits of assays were 4 pg of A. fumigatus gDNA, 2 copies of A. fumigatus and 2 copies of C. albicans respectively. The results of quantitative real-time PCR matched well with each other and were identical to the classical procedures in all patients. Among the 11 patients with invasive fungal disease, the pathogens were identified down to a genus level in 2 invasive aspergillosis patients and 2 invasive candidiasis patients. And the other 7 patients could be diagnosed with invasive fungal disease by the universal fungi assay. All the 10 serum samples from non-invasive fungal disease patients were revealed as negative. CONCLUSION: The universal fungi assay is helpful in screening while the assays of Aspergillus spp. and the Candida spp. may identify the pathogens down to a genus level.

Rapid identification of Pseudallescheria and Scedosporium strains by using rolling circle amplification.

The Pseudallescheria boydii complex, comprising environmental pathogens with Scedosporium anamorphs, has recently been subdivided into five main species: Scedosporium dehoogii, S. aurantiacum, Pseudallescheria minutispora, P. apiosperma, and P. boydii, while the validity of some other taxa is being debated. Several Pseudallescheria and Scedosporium species are indicator organisms of pollution in soil and water. Scedosporium dehoogii in particular is enriched in soils contaminated by aliphatic hydrocarbons. In addition, the fungi may cause life-threatening infections involving the central nervous system in severely impaired patients. For screening purposes, rapid and economic tools for species recognition are needed. Our aim is to establish rolling circle amplification (RCA) as a screening tool for species-specific identification of Pseudallescheria and Scedosporium. With this aim, a set of padlock probes was designed on the basis of the internal transcribed spacer (ITS) region, differing by up to 13 fixed mutations. Padlock probes were unique as judged from sequence comparison by BLAST search in GenBank and in dedicated research databases at CBS (Centraalbureau voor Schimmelcultures Fungal Biodiversity Centre). RCA was applied as an in vitro tool, tested with pure DNA amplified from cultures. The species-specific padlock probes designed in this study yielded 100% specificity. The method presented here was found to be an attractive alternative to identification by restriction fragment length polymorphism (RFLP) or sequencing. The rapidity (<1 day), specificity, and low costs make RCA a promising screening tool for environmentally and clinically relevant fungi.

Identification of Pseudallescheria and Scedosporium species by three molecular methods.

The major clinically relevant species in Scedosporium (teleomorph Pseudallescheria) are Pseudallescheria boydii, Scedosporium aurantiacum, Scedosporium apiospermum, and Scedosporium prolificans, while Pseudallescheria minutispora, Petriellopsis desertorum, and Scedosporium dehoogii are exceptional agents of disease. Three molecular methods targeting the partial ß-tubulin gene were developed and evaluated to identify six closely related species of the S. apiospermum complex using quantitative real-time PCR (qPCR), PCR-based reverse line blot (PCR-RLB), and loop-mediated isothermal amplification (LAMP). qPCR was not specific enough for the identification of all species but had the highest sensitivity. The PCR-RLB assay was efficient for the identification of five species. LAMP distinguished all six species unambiguously. The analytical sensitivities of qPCR, PCR-RLB, and LAMP combined with MagNAPure, CTAB (cetyltrimethylammonium bromide), and FTA filter (Whatman) extraction were 50, 5 × 10(3), and 5 × 10(2) cells/µl, respectively. When LAMP was combined with a simplified DNA extraction method using an FTA filter, identification to the species level was achieved within 2 h, including DNA extraction. The FTA-LAMP assay is therefore recommended as a cost-effective, simple, and rapid method for the identification of Scedosporium species.

[Utility of galactomannan enzyme immunoassay and (1,3)beta-D-glucan assay in invasive fungal infection].

OBJECTIVE: To evaluate the performance of the galactomannan enzyme immunoassay (GM test) and (1,3)beta-D-glucan assay (G test) for the diagnosis of invasive fungal infection (IFI). METHODS: A retrospective study was performed in 115 hospitalized patients at Peking University First Hospital who were at risk of IFI. Patients were diagnosed as IFI according to 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. The sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) were calculated at different cutoff values for two assays respectively. Two tests were combined to evaluate the changes of sensitivity, specificity, PPV and NPV. RESULTS: The best sensitivity (54.5%, 63.6%) and specificity (77.9%, 69.2%) were obtained with the cutoff values of 0.5 and 20 x 10(3) pg/L in GM test and G test respectively. The PPV were 20.7% and 17.9%, and the NPV were 94.2% and 94.7% respectively. The sensitivity increased to 90.9% and the specificity was 52.9% after a combined utility of two tests. CONCLUSION: The GM test and G tests are both useful in diagnosis of IFI with the cutoff values of 0.5 and 20 x 10(3) pg/L. A better sensitivity is acquired if there is a combined utility of two tests.

In vitro combined activity of amphotericin B, caspofungin and voriconazole against clinical isolates of Trichosporon asahii.

Disseminated infections caused by Trichosporon asahii are difficult to resolve. Combination regimens with synergistic drugs could provide additional options for treating trichosporonosis. The aim of this study was to evaluate the antifungal activities of voriconazole (VCZ), caspofungin (CAS) and amphotericin B (AMB) alone or in combination in vitro against clinical isolates of T. asahii. The combined antifungal activities of VCZ, CAS and AMB against 18 clinical isolates were assessed by a chequerboard microdilution method. CAS combined with AMB showed the highest percentage of synergistic effects (89%), much higher than those of the other two combinations (AMB/VCZ and CAS/VCZ both 17%). No antagonistic effect was observed in any case. This study demonstrates that the activity of two combined antifungals, especially the combination of CAS and AMB, against T. asahii is more effective than that of a drug alone against this fungus, suggesting that combined antifungal therapy may be a potential strategy for treating disseminated trichosporonosis.

Cyclophosphamide reduces dectin-1 expression in the lungs of naive and Aspergillus fumigatus-infected mice.

Aspergillus fumigatus is an important opportunistic fungal pathogen. Patients treated with chemotherapeutic agent such as cyclophosphamide are susceptible to invasive pulmonary aspergillosis. Dectin-1 is a pattern recognition receptor critically involved in immune responses to A. fumigatus. Therefore, we tested whether cyclophosphamide treatment could cause alterations in dectin-1 expression in the lung, which could contribute to invasive pulmonary Aspergillus infections in patients. We established a murine A. fumigatus infectious model to investigate the kinetics of dectin-1 expression in lung tissues in the presence or absence of cyclophosphamide treatment. During infection, dectin-1 expression was strikingly increased in immunocompetent mice infected with A. fumigatus as compared to those in a non-infected control group. In vitro macrophages stimulated with heat-inactivated A. fumigatus conidia expressed a significantly elevated level of dectin-1. Infected mice treated with cyclophosphamide showed decreased levels of dectin-1 and a higher fungal burden in the lung than the infected mice without cyclophosphamide treatment. These results suggest that dectin-1 is involved in host defense against A. fumigatus infection and that suppression of dectin-1 expression caused by cyclophosphamide may contribute to susceptibility to infections caused by this fungus in the immunocompromised host.