Journal: Journal of Clinical Microbiology

Article Title: Design and Validation of Transcription-Mediated-Amplification Nucleic Acid Amplification Tests for Mycoplasma genitalium

doi: 10.1128/JCM.00264-19

Figure Lengend Snippet: Analytical specificity (cross-reactivity) of TMA assays for Mycoplasma genitalium

Article Snippet: All tests were negative for all microorganisms, including closely related species Mycoplasma pneumoniae and Mycoplasma hominis . table ft1 table-wrap mode="anchored" t5 TABLE 2 caption a7 Cross-reactivity panel ATCC or Hologic lot no. Microorganism AMG IVD Alt TMA-1 Alt TMA-2 Alt TMA-3 % pos a Avg S/CO b % pos Avg S/CO % pos Avg S/CO % pos Avg S/CO 1 15309 Acinetobacter lwoffii 0 0 0 0.10 0 0.06 0 0.05 12102 Actinomyces israelii 8750 Alcaligenes faecalis BAA-55 Atopobium vaginae 2 25285 Bacteroides fragilis 0 0 0 0.07 0 0.07 0 0.06 15703 Bifidobacterium adolescentis 33560 Campylobacter jejuni 18804 Candida albicans 3 VR-878 Chlamydia trachomatis 0 0 0 0.04 0 0.06 0 0.09 9689 Clostridium difficile 33030 Corynebacterium genitalium 32045 Cryptococcus neoformans 4 VR-2356 Cytomegalovirus 0 0 0 0.05 0 0.09 0 0.05 13047 Enterobacter cloacae 19433 Enterococcus faecalis 11775 Escherichia coli 5 25586 Fusobacterium nucleatum 0 0 0 0.03 0 0.07 0 0.05 14018 Gardnerella vaginalis 33940 Haemophilus ducreyi 6 VR-539 Herpes simplex virus I 0 0 0 0.05 0 0.09 0 0.05 VR-540 Herpes simplex virus II 115500 HIV-1 7 23357 Klebsiella pneumoniae 0 0 0 0.07 0 0.05 0 0.05 4356 Lactobacillus acidophilus 33820 Lactobacillus crispatus 14201 Leptotrichia buccalis 8 15313 Listeria monocytogenes 0 0 0 0.04 0 0.07 0 0.05 35241 Mobiluncus curtisii 19424 Neisseria gonorrhoeae 14955 Finegoldia magna 9 29303 Prevotella bivia 0 0 0 0.05 0 0.04 0 0.04 6919 Propionibacterium acnes 8427 Proteus vulgaris 10145 Pseudomonas aeruginosa 10 10145 Staphylococcus aureus 0 0 0 0.07 0 0.07 0 0.05 14990 Staphylococcus epidermis 13813 Streptococcus agalactiae 12344 Streptococcus pyogenes 11 30236 Trichomonas vaginalis 0 0 0 0.09 0 0.07 0 0.04 27813 Ureaplasma parvum 27618 Ureaplasma urealyticum 12 23114 Mycoplasma hominis 0 0 0 0.06 0 0.06 0 0.05 13 29342 Mycoplasma pneumoniae 0 0.26 0 0.53 0 0.13 0 0.06 14 NA c HPV 6 0 0 0 0.06 0 0.07 0 0.05 NA HPV 11 HTB-35 HPV 16 (SiHa cells) CCL-2 HPV 18 (HeLa cells) NA HPV 58 NA HPV 39 NA HPV 51 15 29666 Chromobacterium violaceum 0 0 0 0.04 0 0.08 0 0.05 13253 Elizabethkingia meningoseptica 16 30000 Pentatrichomonas hominis 0 0 0 0.10 0 0.09 0 0.05 NA Megasphaera type 1 17 NA No microorganism control (matrix only) 0 0 0 0.04 0 0.04 0 0.05 18 43867 Staphylococcus saprophyticus 0 0 0 0.05 0 0.06 0 0.06 DSM 25311 Mobiluncus mulieris Open in a separate window a pos, positive. b S/CO, signal-to-cutoff ratio. c NA, not applicable.

Techniques:

Journal: Antimicrobial Agents and Chemotherapy

Article Title: Fungicidal Potency and Mechanisms of θ-Defensins against Multidrug-Resistant Candida Species

doi: 10.1128/AAC.00111-18

Figure Lengend Snippet: Fungicidal activities of RTD-1 and Hst 5 against drug-sensitive and MDR C. albicans. Drug-sensitive (ATCC 18804) and MDR (ATCC 64124) C. albicans strains were incubated with indicated concentrations of RTD-1 or 100 μg/ml Hst 5 (H) for 60 min in PG buffer for 0 to 60 min. Fungicidal activity was quantified by CFU quantitation after 24 h of incubation. Experiments were repeated twice using three biological replicates, and error bars are shown.

Article Snippet: In the presence of divalent cation-containing media, killing of ATCC 18804 required higher RTD concentrations than with PG buffer, consistent with a previous study employing C. albicans laboratory strain 16820 ( 39 ).

Techniques: Incubation, Activity Assay, Quantitation Assay

Journal: Antimicrobial Agents and Chemotherapy

Article Title: Fungicidal Potency and Mechanisms of θ-Defensins against Multidrug-Resistant Candida Species

doi: 10.1128/AAC.00111-18

Figure Lengend Snippet: RTD-1 rapidly permeabilizes C. albicans. PI uptake by C. albicans strains ATCC 18804 (18804) (A) and ATCC 64124 (64124) (B) was monitored by confocal microscopy after the addition of 5 μg/ml RTD-1 or 94 μg/ml Hst 5 for 20 min (left). Graphs on the right show the time course of mean PI fluorescence intensity. Data are representative of the results from an individual experiment that was performed three times.

Article Snippet: In the presence of divalent cation-containing media, killing of ATCC 18804 required higher RTD concentrations than with PG buffer, consistent with a previous study employing C. albicans laboratory strain 16820 ( 39 ).

Techniques: Confocal Microscopy, Fluorescence

Journal: Antimicrobial Agents and Chemotherapy

Article Title: Fungicidal Potency and Mechanisms of θ-Defensins against Multidrug-Resistant Candida Species

doi: 10.1128/AAC.00111-18

Figure Lengend Snippet: RTD-1 induces rapid release of ATP from C. albicans. (A) C. albicans ATCC 18804 cells were incubated with 0.5 μg/ml RTD-1 or 94 μg/ml histatin-5 in a 0.1-ml reaction mixture volume and monitored for ATP release. Incubations were carried out in the absence or presence of 1 mM sodium azide. (B) Replicates of the incubation mixtures in panel A were analyzed for fungicidal activity, demonstrating that the killing of RTD-1 is rapid and not ATP dependent. Data are representative of an individual experiment repeated twice. (C) RTD-1 (0.5 μg/ml) causes rapid ATP release from MDR C. albicans ATCC 64124 cells, whereas 94 μg/ml Hst 5 causes much more gradual ATP release. The reduction of ATP levels observed at 90 min in RTD-1 containing incubations was dose-dependently reversed by ATPase inhibitor concanamycin A (see the text). The experiments shown in panels A and C were repeated twice using three biological replicates, and error bars are shown.

Article Snippet: In the presence of divalent cation-containing media, killing of ATCC 18804 required higher RTD concentrations than with PG buffer, consistent with a previous study employing C. albicans laboratory strain 16820 ( 39 ).

Techniques: Incubation, Activity Assay

Journal: Antimicrobial Agents and Chemotherapy

Article Title: Fungicidal Potency and Mechanisms of θ-Defensins against Multidrug-Resistant Candida Species

doi: 10.1128/AAC.00111-18

Figure Lengend Snippet: RTD-1 rapidly induces intracellular ROS in C. albicans. DCFH-DA fluorescence was measured in drug-sensitive C. albicans ATCC 18804 and MDR ATCC 64124 after incubation with 6.25 μg/ml RTD-1 for the indicated intervals. The significance of the effect of RTD-1 treatment at all time points was a P value of <0.0001 (*; 2-tailed Student's t test). Experiments were performed using three biological replicates and repeated three times.

Article Snippet: In the presence of divalent cation-containing media, killing of ATCC 18804 required higher RTD concentrations than with PG buffer, consistent with a previous study employing C. albicans laboratory strain 16820 ( 39 ).

Techniques: Fluorescence, Incubation

Journal: Antimicrobial Agents and Chemotherapy

Article Title: Fungicidal Potency and Mechanisms of θ-Defensins against Multidrug-Resistant Candida Species

doi: 10.1128/AAC.00111-18

Figure Lengend Snippet: Comparative stability of RTD-1 and Hst 5 in C. albicans cells lysates. Five micrograms of RTD-1 or Hst 5 was incubated with lysates of C. albicans cells (strains ATCC 64124, ATCC 18804, and ATCC MYA-2876) and analyzed on acetic acid-urea (AU)-PAGE at time zero and after 2 h of incubation. One-fifth of each incubation mixture (1.0 μg input) was loaded in each well; control (ctrl) lanes were loaded with 1.0 μg of untreated RTD-1 or Hst 5. The gel was stained with formalin-Coomassie blue (49). Data are representative of experiments repeated twice.

Article Snippet: In the presence of divalent cation-containing media, killing of ATCC 18804 required higher RTD concentrations than with PG buffer, consistent with a previous study employing C. albicans laboratory strain 16820 ( 39 ).

Techniques: Incubation, Staining

Journal: Antimicrobial Agents and Chemotherapy

Article Title: Fungicidal Potency and Mechanisms of θ-Defensins against Multidrug-Resistant Candida Species

doi: 10.1128/AAC.00111-18

Figure Lengend Snippet: MICs of θ-defensins and clinical antifungals a

Article Snippet: In the presence of divalent cation-containing media, killing of ATCC 18804 required higher RTD concentrations than with PG buffer, consistent with a previous study employing C. albicans laboratory strain 16820 ( 39 ).

Techniques:

Journal: Antimicrobial Agents and Chemotherapy

Article Title: Fungicidal Potency and Mechanisms of θ-Defensins against Multidrug-Resistant Candida Species

doi: 10.1128/AAC.00111-18

Figure Lengend Snippet: MFCs of θ-defensins and clinical antifungals a

Article Snippet: In the presence of divalent cation-containing media, killing of ATCC 18804 required higher RTD concentrations than with PG buffer, consistent with a previous study employing C. albicans laboratory strain 16820 ( 39 ).

Techniques: