Journal: Biomolecules

Article Title: Hybrid Imaging of Aspergillus fumigatus Pulmonary Infection with Fluorescent, 68 Ga-Labelled Siderophores

doi: 10.3390/biom10020168

Figure Lengend Snippet: Growth of A. fumigatus mutant strain Δ sidA/ Δ ftrA after 48 h incubation at 37 °C on iron-depleted Aspergillus minimal medium agar with different iron containing fluorophore conjugates. Growth is reflected by whitish-mycelia, while sporulation is reflected by the green colour, which arises from the green conidial-specific pigment. The last row shows controls of agar without siderophores: W = sterile water; S = Spores.

Article Snippet: A. fumigatus (ATCC 46645) was cultivated for 14 h and A. terreus (ATCC 3633) for 48 h to obtain well developed germlings and young hyphae without extensive germling or vegetative hyphal fusion.

Techniques: Mutagenesis, Incubation, Sterility

Journal: Biomolecules

Article Title: Hybrid Imaging of Aspergillus fumigatus Pulmonary Infection with Fluorescent, 68 Ga-Labelled Siderophores

doi: 10.3390/biom10020168

Figure Lengend Snippet: Fluorescence microscopy of [Fe]DAFC-Cy5 and [Fe]DAFC-SulfoCy5 in A. fumigatus and A. terreus . ( A ) [Fe]DAFC-Cy5 labels what appear to be tubular vacuoles with a clear concentration in hyphal tips. ( B ) Incubation with Cy5 carboxylic acid “dye alone” results in a very similar labelling pattern as [Fe]DAFC-Cy5. ( C , D ) In contrast, fluorescence microscopy does not visualize any uptake of [Fe]DAFC-SulfoCy5 and -SulfoCy5 “dye alone” by A. fumigatus . ( E ) A. terreus , which lacks a MirB homologous transporter and consequently TAFC uptake, does not show uptake of [Fe]DAFC-SulfoCy5, while it internalizes Cy5 carboxylic acid “dye alone” ( F ). ( G , H ) A. terreus does not internalize [Fe]DAFC-SulfoCy5 or -SulfoCy5 “dye alone”. Scale bars, 10 μm.

Article Snippet: A. fumigatus (ATCC 46645) was cultivated for 14 h and A. terreus (ATCC 3633) for 48 h to obtain well developed germlings and young hyphae without extensive germling or vegetative hyphal fusion.

Techniques: Fluorescence, Microscopy, Concentration Assay, Incubation

Journal: Biomolecules

Article Title: Hybrid Imaging of Aspergillus fumigatus Pulmonary Infection with Fluorescent, 68 Ga-Labelled Siderophores

doi: 10.3390/biom10020168

Figure Lengend Snippet: Coronal µPET/CT slices of A. fumigatus infected (top row) and non-infected animals (bottom row) 45 min p.i. in immunocompromised Lewis rats (approx. 5–10 MBq injected dose).

Article Snippet: A. fumigatus (ATCC 46645) was cultivated for 14 h and A. terreus (ATCC 3633) for 48 h to obtain well developed germlings and young hyphae without extensive germling or vegetative hyphal fusion.

Techniques: Infection, Injection

Journal: Biomolecules

Article Title: Hybrid Imaging of Aspergillus fumigatus Pulmonary Infection with Fluorescent, 68 Ga-Labelled Siderophores

doi: 10.3390/biom10020168

Figure Lengend Snippet: Fluorescence images of lungs excised from A. fumigatus infected and non-infected Lewis rats 1.5 h after the injection of different 68 Ga-labelled fluorophore conjugates.

Article Snippet: A. fumigatus (ATCC 46645) was cultivated for 14 h and A. terreus (ATCC 3633) for 48 h to obtain well developed germlings and young hyphae without extensive germling or vegetative hyphal fusion.

Techniques: Fluorescence, Infection, Injection

Journal: Frontiers in Immunology

Article Title: Neutrophil-Specific Knockdown of β2 Integrins Impairs Antifungal Effector Functions and Aggravates the Course of Invasive Pulmonal Aspergillosis

doi: 10.3389/fimmu.2022.823121

Figure Lengend Snippet: Infection with IPA caused an impaired survival (A) and an aggravated course of the disease (B) in CD18 Ly6G cKO as compared to CD18 fl/fl mice. CD18 fl/fl and CD18 Ly6G cKO mice were infected i.t. with A fumigatus (each 10 7 conidia/mouse) in 2 independent experiments. (A) Survival was monitored daily for 2 weeks and is presented in a Kaplan-Meier survival curve. In parallel settings PMN were depleted in some mice via injection of an anti-Gr-1 antibody one day before infection. Data show the cumulative results of two independent experiments with a total of 12 (CD18 fl/fl ) and 13 (CD18 Ly6G cKO) mice/group. 5 mice/group received an anti-Gr-1 antibody in order to deplete PMN in these mice. All Gr-1 depleted mice died within the first days after IPA infection, whereas all non-depleted CD18 fl/fl mice survived. By contrast, some non-depleted CD18 Ly6G cKO mice (n=2) deceased within the first week after IPA infection. (B) The clinical course of IPA of monitoring was assessed in CD18 fl/fl (n=7) and CD18 Ly6G cKO mice (n=8) for 14 days. Parameters comprised breathing, reaction to pain overall appearance, hypothermia, strong weight loss, motoric disabilities and apathy (each 0-2).

Article Snippet: Mice were anesthetized with 14.5% Ketamin (50mg/ml)/5.7% Xylazin (0.2%) and were subsequently challenged with 10 7 A. fumigatus conidia (strain ATCC 46645) applied intratracheally as described ( , ).

Techniques: Infection, Injection

Journal: Frontiers in Immunology

Article Title: Neutrophil-Specific Knockdown of β2 Integrins Impairs Antifungal Effector Functions and Aggravates the Course of Invasive Pulmonal Aspergillosis

doi: 10.3389/fimmu.2022.823121

Figure Lengend Snippet: CD18 Ly6G cKO mice show a higher pulmonary fungal burden. Histopathological analysis of H&E and Grocott-stained lungs derived from IPA-infected mice 24h upon A.fumigatus inoculation revealed a higher fungal burden and a stronger lung damage (i.e., hyaline membranes, fibrin-exudate within the alveoli) in CD18 Ly6G cKO mice (A) . Cellular inflammation did not show significant genotype-dependent differences. Representative examples of histological analysis are shown in (B) (Magnification 10x). Data in (A) denote results of histopathological analysis of n=9-10 mice/genotype. We further observed higher CFU counts in serial dilutions of lung homogenates (1:500) after incubation for 24h on Sabouraud-4% Glucose agar plates (C) . Data show the mean ± SEM of 6 mice/group. Statistically significant differences between groups are indicated (*p<0.05, **p<0.005, ***p<0.001).

Article Snippet: Mice were anesthetized with 14.5% Ketamin (50mg/ml)/5.7% Xylazin (0.2%) and were subsequently challenged with 10 7 A. fumigatus conidia (strain ATCC 46645) applied intratracheally as described ( , ).

Techniques: Staining, Derivative Assay, Infection, Incubation

Journal: Frontiers in Immunology

Article Title: Neutrophil-Specific Knockdown of β2 Integrins Impairs Antifungal Effector Functions and Aggravates the Course of Invasive Pulmonal Aspergillosis

doi: 10.3389/fimmu.2022.823121

Figure Lengend Snippet: The BAL fluid of A. fumigatus infected CD18 Ly6G cKO mice contains lower levels of proinflammatory cytokines. CD18 fl/fl and CD18 Ly6G cKO mice were infected i.t. with A. fumigatus . On the next day, mice were euthanized, and cytokines in BAL fluid were analyzed. Data denote the mean ± SEM of 6-10 samples analyzed per group. Statistically significant differences between groups are indicated (* p < 0.05).

Article Snippet: Mice were anesthetized with 14.5% Ketamin (50mg/ml)/5.7% Xylazin (0.2%) and were subsequently challenged with 10 7 A. fumigatus conidia (strain ATCC 46645) applied intratracheally as described ( , ).

Techniques: Infection

Journal: Frontiers in Immunology

Article Title: Neutrophil-Specific Knockdown of β2 Integrins Impairs Antifungal Effector Functions and Aggravates the Course of Invasive Pulmonal Aspergillosis

doi: 10.3389/fimmu.2022.823121

Figure Lengend Snippet: CD18 Ly6G cKO mice infected with A. fumigatus are characterized by elevated lung infiltration of PMN, but not in spleens and blood (n=10/genotype) (A) . Elevated PMN counts have also been found in Cytospins (bars depict the mean ± SEM of n=8 cytospins/genotype; cell infiltration has been assessed using a scoring system; 0=missing - 3=strongest infiltration) (B) . Here, we could additionally observe higher numbers of mononuclear cells (B) . This is consistent with the finding of higher macrophage counts in the BALF of CD18 fl/fl mice as compared to CD18 Ly6G cKO mice observed in FACS-analysis (B) . Assessment of PMN apoptosis revealed a stronger expression of apoptosis marker Annexin V in PMN derived from BALF and blood of CD18 Ly6G cKO mice (C) . Bars depict the mean ± SEM of the relative cell counts found in n=8 cytospins/genotype. Legend in A applies to all panels. Statistically significant differences between groups are indicated (*p<0.05)

Article Snippet: Mice were anesthetized with 14.5% Ketamin (50mg/ml)/5.7% Xylazin (0.2%) and were subsequently challenged with 10 7 A. fumigatus conidia (strain ATCC 46645) applied intratracheally as described ( , ).

Techniques: Infection, Expressing, Marker, Derivative Assay

Journal: Frontiers in Immunology

Article Title: Neutrophil-Specific Knockdown of β2 Integrins Impairs Antifungal Effector Functions and Aggravates the Course of Invasive Pulmonal Aspergillosis

doi: 10.3389/fimmu.2022.823121

Figure Lengend Snippet: Phagocytosis of inert particles and fungal conidia is less effective in PMN of CD18 Ly6G cKO mice. Freshly isolated PMN were co-incubated either with nanoparticles (NP), microBeads (A) or with GFP-fluorescent A. fumigatus conidia (AFC) at 37°C with indicated ratios (B) . Simultaneous co-incubation at 4°C served to differentiate mere adhesion from temperature-dependent binding. After 30 min and 60 min the frequency of either Cy5 positive NP, PE-positive microBeads (A) , or GFP-positive PMN (B) was determined by flow cytometry. Data represent the mean ± SEM of 3 samples analyzed/group. Exemplary flow cytometry data depicting the diminished uptake of GFP-fluorescent conidia by PMN from CD18 Ly6G cKO mice are shown in (C) . Statistically significant differences between groups are indicated (*p<0.05, **p<0.005).

Article Snippet: Mice were anesthetized with 14.5% Ketamin (50mg/ml)/5.7% Xylazin (0.2%) and were subsequently challenged with 10 7 A. fumigatus conidia (strain ATCC 46645) applied intratracheally as described ( , ).

Techniques: Isolation, Incubation, Binding Assay, Flow Cytometry

Journal: Frontiers in Immunology

Article Title: Neutrophil-Specific Knockdown of β2 Integrins Impairs Antifungal Effector Functions and Aggravates the Course of Invasive Pulmonal Aspergillosis

doi: 10.3389/fimmu.2022.823121

Figure Lengend Snippet: Impaired oxidative and non-oxidative effector functions of CD18-deficient PMN. We could observe that the formation of NET after 3h of incubation (A) and ROS-generation (B) were significantly lower in PMN derived from CD18 Ly6G cKO mice, particularly after stimulation with A. fumigatus conidia. Data represent the mean ± SEM of 3 samples analyzed/group. Statistically significant differences between groups are indicated (*p<0.05)

Article Snippet: Mice were anesthetized with 14.5% Ketamin (50mg/ml)/5.7% Xylazin (0.2%) and were subsequently challenged with 10 7 A. fumigatus conidia (strain ATCC 46645) applied intratracheally as described ( , ).

Techniques: Incubation, Derivative Assay

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: New Claims for Wild Carrot ( Daucus carota subsp. carota ) Essential Oil

doi: 10.1155/2016/9045196

Figure Lengend Snippet: Antifungal activity (MIC and MLC) of Daucus carota subsp. carota essential oil for Candida spp., Cryptococcus neoformans , dermatophyte, and Aspergillus strains.

Article Snippet: The antifungal properties of the essential oil were tested against three Candida reference strains ( C. albicans ATCC 10231, C. tropicalis ATCC 13803, and C. parapsilosis ATCC 90018) and two clinical strains ( C. krusei H9 and C. guilliermondii MAT23); one Cryptococcus neoformans reference strain ( C. neoformans CECT 1078); four dermatophyte strains ( Trichophyton rubrum CECT 2794, T. mentagrophytes var. interdigitale CECT 2958, T. verrucosum CECT 2992, and Microsporum gypseum CECT 2908); the remaining dermatophytes were clinically isolated ( T. mentagrophytes FF7, M. canis FF1, and Epidermophyton floccosum FF9); two reference Aspergillus strains ( A. niger ATCC 16404 and A. fumigatus ATCC 46645); and one Aspergillus strain was from a clinical origin ( A. flavus F44).

Techniques: Activity Assay

Journal:

Article Title: Molecular Basis for Fungal Selectivity of Novel Antimitotic Compounds

doi: 10.1128/AAC.47.7.2273-2282.2003

Figure Lengend Snippet: Strains used in this study

Article Snippet: Yeast transformation and plasmid selection were performed using standard methods ( 1 ). table ft1 table-wrap mode="anchored" t5 TABLE 1. caption a7 Strain a Genotype Origin (reference) AF46645 Wild type 46645, American Type Culture Collection SC5314 Wild type SC5314 (7) SC760 MAT a ura3-52 lys2 trp1 Δ1 F760, Gerald Fink, Whitehead Institute SC1210 MAT α ura3-52 lys2-801 ade2-101 his3Δ200 leu2Δ1 YP102, Phil Hieter, Univ.

Techniques: Mutagenesis, Selection

Journal:

Article Title: Molecular Basis for Fungal Selectivity of Novel Antimitotic Compounds

doi: 10.1128/AAC.47.7.2273-2282.2003

Figure Lengend Snippet: MC-305,904 analog thiazoline ring substituent effects on activity toward pathogenic fungal species

Article Snippet: Yeast transformation and plasmid selection were performed using standard methods ( 1 ). table ft1 table-wrap mode="anchored" t5 TABLE 1. caption a7 Strain a Genotype Origin (reference) AF46645 Wild type 46645, American Type Culture Collection SC5314 Wild type SC5314 (7) SC760 MAT a ura3-52 lys2 trp1 Δ1 F760, Gerald Fink, Whitehead Institute SC1210 MAT α ura3-52 lys2-801 ade2-101 his3Δ200 leu2Δ1 YP102, Phil Hieter, Univ.

Techniques: Activity Assay, Mutagenesis