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Article ; Online: Epidemiology and drug susceptibility of nontuberculous mycobacteria (NTM) in Italy in 2016-2020.

Giannoni, Federico / Lanni, Alessio / Iacobino, Angelo / Fattorini, Lanfranco

Annali dell'Istituto superiore di sanita

2023 Volume 59, Issue 2, Page(s) 132–138

Abstract: Introduction: Nontuberculous mycobacteria (NTM) are environmental mycobacteria which may cause pulmonary and extrapulmonary diseases. These organisms are difficult to treat due to their intrinsic drug-resistance. In Italy, no major nationwide study on ... ...

Abstract	Introduction: Nontuberculous mycobacteria (NTM) are environmental mycobacteria which may cause pulmonary and extrapulmonary diseases. These organisms are difficult to treat due to their intrinsic drug-resistance. In Italy, no major nationwide study on NTM epidemiology and drug susceptibility was performed. Methods: Data on the epidemiology of 7,469 NTM clinical isolates identified in Italy in 2016-2020 and on the minimum inhibitory concentrations (MICs) of 1,506 of these strains were analysed. Results: Overall, 63 species were identified in 42 hospital laboratories located in 16 out of 20 regions, with Mycobacterium avium complex (MAC) being the most frequently isolated, followed by M. gordonae, M. xenopi, M. abscessus. The MICs of 12 drugs for MAC, M. xenopi, M. kansasii, M. abscessus, M. fortuitum and M. chelonae were interpreted for clinical significance (susceptible, intermediate, resistant) based on the guidelines published by the Clinical and Laboratory Standards Institute in November 2018. Conclusions: Our data are in line with other nationwide studies and may be of value for further update of microbiological and clinical guidelines.
MeSH term(s)	Humans ; Nontuberculous Mycobacteria ; Mycobacterium Infections, Nontuberculous/epidemiology ; Mycobacterium Infections, Nontuberculous/microbiology ; Microbial Sensitivity Tests ; Italy/epidemiology
Language	English
Publishing date	2023-02-17
Publishing country	Italy
Document type	Journal Article
ZDB-ID	950344-4
ISSN	2384-8553 ; 0021-2571
ISSN (online)	2384-8553
ISSN	0021-2571
DOI	10.4415/ANN_23_02_06
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Article: Eradication of Drug-Tolerant

Lanni, Alessio / Iacobino, Angelo / Fattorini, Lanfranco / Giannoni, Federico

Microorganisms

2023 Volume 11, Issue 6

Abstract: The lungs of tuberculosis (TB) patients contain a spectrum of granulomatous lesions, ranging from solid and well-vascularized cellular granulomas to avascular caseous granulomas. In solid granulomas, current therapy kills actively replicating (AR) ... ...

Abstract	The lungs of tuberculosis (TB) patients contain a spectrum of granulomatous lesions, ranging from solid and well-vascularized cellular granulomas to avascular caseous granulomas. In solid granulomas, current therapy kills actively replicating (AR) intracellular bacilli, while in low-vascularized caseous granulomas the low-oxygen tension stimulates aerobic and microaerophilic AR bacilli to transit into non-replicating (NR), drug-tolerant and extracellular stages. These stages, which do not have genetic mutations and are often referred to as persisters, are difficult to eradicate due to low drug penetration inside the caseum and mycobacterial cell walls. The sputum of TB patients also contains viable bacilli called differentially detectable (DD) cells that, unlike persisters, grow in liquid, but not in solid media. This review provides a comprehensive update on drug combinations killing in vitro AR and drug-tolerant bacilli (persisters and DD cells), and sterilizing
Language	English
Publishing date	2023-06-06
Publishing country	Switzerland
Document type	Journal Article ; Review
ZDB-ID	2720891-6
ISSN	2076-2607
ISSN	2076-2607
DOI	10.3390/microorganisms11061511
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Article: Activity of Drug Combinations against Mycobacterium abscessus Grown in Aerobic and Hypoxic Conditions

Lanni, Alessio / Borroni, Emanuele / Iacobino, Angelo / Russo, Cristina / Gentile, Leonarda / Fattorini, Lanfranco / Giannoni, Federico

Microorganisms. 2022 July 14, v. 10, no. 7

2022

Abstract: Infections caused by Mycobacterium abscessus (Mab), an environmental non-tuberculous mycobacterium, are difficult to eradicate from patients with pulmonary diseases such as cystic fibrosis and bronchiectasis even after years of antibiotic treatments. In ... ...

Abstract	Infections caused by Mycobacterium abscessus (Mab), an environmental non-tuberculous mycobacterium, are difficult to eradicate from patients with pulmonary diseases such as cystic fibrosis and bronchiectasis even after years of antibiotic treatments. In these people, the low oxygen pressure in mucus and biofilm may restrict Mab growth from actively replicating aerobic (A) to non-replicating hypoxic (H) stages, which are known to be extremely drug-tolerant. After the exposure of Mab A and H cells to drugs, killing was monitored by measuring colony-forming units (CFU) and regrowth in liquid medium (MGIT 960) of 1-day-old A cells (A1) and 5-day-old H cells (H5). Mab killing was defined as a lack of regrowth of drug-exposed cells in MGIT tubes after >50 days of incubation. Out of 18 drugs tested, 14-day treatments with bedaquiline-amikacin (BDQ-AMK)-containing three-drug combinations were very active against A1 + H5 cells. However, drug-tolerant cells (persisters) were not killed, as shown by CFU curves with typical bimodal trends. Instead, 56-day treatments with the nitrocompounds containing combinations BDQ-AMK-rifabutin-clarithromycin-nimorazole and BDQ-AMK-rifabutin-clarithromycin-metronidazole-colistin killed all A1 + H5 Mab cells in 42 and 56 days, respectively, as shown by lack of regrowth in agar and MGIT medium. Overall, these data indicated that Mab persisters may be killed by appropriate drug combinations.
Keywords	Mycobacterium abscessus ; agar ; antibiotics ; biofilm ; cystic fibrosis ; liquids ; mucus ; oxygen ; people ; regrowth
Language	English
Dates of publication	2022-0714
Publishing place	Multidisciplinary Digital Publishing Institute
Document type	Article
ZDB-ID	2720891-6
ISSN	2076-2607
ISSN	2076-2607
DOI	10.3390/microorganisms10071421
Database	NAL-Catalogue (AGRICOLA)

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Article: Moxifloxacin Activates the SOS Response in

Iacobino, Angelo / Piccaro, Giovanni / Pardini, Manuela / Fattorini, Lanfranco / Giannoni, Federico

Microorganisms

2021 Volume 9, Issue 2

Abstract: Previous studies ... ...

Abstract	Previous studies on
Language	English
Publishing date	2021-01-27
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2720891-6
ISSN	2076-2607
ISSN	2076-2607
DOI	10.3390/microorganisms9020255
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Article: Activity of Drug Combinations against

Lanni, Alessio / Borroni, Emanuele / Iacobino, Angelo / Russo, Cristina / Gentile, Leonarda / Fattorini, Lanfranco / Giannoni, Federico

Microorganisms

2022 Volume 10, Issue 7

Abstract	Infections caused by Mycobacterium abscessus (Mab), an environmental non-tuberculous mycobacterium, are difficult to eradicate from patients with pulmonary diseases such as cystic fibrosis and bronchiectasis even after years of antibiotic treatments. In these people, the low oxygen pressure in mucus and biofilm may restrict Mab growth from actively replicating aerobic (A) to non-replicating hypoxic (H) stages, which are known to be extremely drug-tolerant. After the exposure of Mab A and H cells to drugs, killing was monitored by measuring colony-forming units (CFU) and regrowth in liquid medium (MGIT 960) of 1-day-old A cells (A1) and 5-day-old H cells (H5). Mab killing was defined as a lack of regrowth of drug-exposed cells in MGIT tubes after >50 days of incubation. Out of 18 drugs tested, 14-day treatments with bedaquiline-amikacin (BDQ-AMK)-containing three-drug combinations were very active against A1 + H5 cells. However, drug-tolerant cells (persisters) were not killed, as shown by CFU curves with typical bimodal trends. Instead, 56-day treatments with the nitrocompounds containing combinations BDQ-AMK-rifabutin-clarithromycin-nimorazole and BDQ-AMK-rifabutin-clarithromycin-metronidazole-colistin killed all A1 + H5 Mab cells in 42 and 56 days, respectively, as shown by lack of regrowth in agar and MGIT medium. Overall, these data indicated that Mab persisters may be killed by appropriate drug combinations.
Language	English
Publishing date	2022-07-14
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2720891-6
ISSN	2076-2607
ISSN	2076-2607
DOI	10.3390/microorganisms10071421
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Article: Moxifloxacin Activates the SOS Response in Mycobacterium tuberculosis in a Dose- and Time-Dependent Manner

Iacobino, Angelo / Piccaro, Giovanni / Pardini, Manuela / Fattorini, Lanfranco / Giannoni, Federico

Microorganisms. 2021 Jan. 27, v. 9, no. 2

2021

Abstract: Previous studies on Escherichia coli demonstrated that sub-minimum inhibitory concentration (MIC) of fluoroquinolones induced the SOS response, increasing drug tolerance. We characterized the transcriptional response to moxifloxacin in Mycobacterium ... ...

Abstract	Previous studies on Escherichia coli demonstrated that sub-minimum inhibitory concentration (MIC) of fluoroquinolones induced the SOS response, increasing drug tolerance. We characterized the transcriptional response to moxifloxacin in Mycobacterium tuberculosis. Reference strain H37Rv was treated with moxifloxacin and gene expression studied by qRT-PCR. Five SOS regulon genes, recA, lexA, dnaE2, Rv3074 and Rv3776, were induced in a dose- and time-dependent manner. A range of moxifloxacin concentrations induced recA, with a peak observed at 2 × MIC (0.25 μg/mL) after 16 h. Another seven SOS responses and three DNA repair genes were significantly induced by moxifloxacin. Induction of recA by moxifloxacin was higher in log-phase than in early- and stationary-phase cells, and absent in dormant bacilli. Furthermore, in an H37Rv fluoroquinolone-resistant mutant carrying the D94G mutation in the gyrA gene, the SOS response was induced at drug concentrations higher than the mutant MIC value. The 2 × MIC of moxifloxacin determined no significant changes in gene expression in a panel of 32 genes, except for up-regulation of the relK toxin and of Rv3290c and Rv2517c, two persistence-related genes. Overall, our data show that activation of the SOS response by moxifloxacin, a likely link to increased mutation rate and persister formation, is time, dose, physiological state and, possibly, MIC dependent.
Keywords	DNA repair ; Escherichia coli ; Mycobacterium tuberculosis ; drug resistance ; drugs ; gene expression ; moxifloxacin ; mutants ; mutation ; mutation rate ; physiological state ; regulon ; toxins ; transcription (genetics)
Language	English
Dates of publication	2021-0127
Publishing place	Multidisciplinary Digital Publishing Institute
Document type	Article
ZDB-ID	2720891-6
ISSN	2076-2607
ISSN	2076-2607
DOI	10.3390/microorganisms9020255
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Article: Activity of DNA-targeted C8-linked pyrrolobenzodiazepine-heterocyclic polyamide conjugates against aerobically and hypoxically grown Mycobacterium tuberculosis under acidic and neutral conditions.

Iacobino, Angelo / Giannoni, Federico / Fattorini, Lanfranco / Brucoli, Federico

The Journal of antibiotics

2018 Volume 71, Issue 9, Page(s) 831–834

Abstract: Mycobacterium tuberculosis (Mtb) is the aetiological agent of tuberculosis, the leading cause of death worldwide from a single infectious agent. Mtb is a highly adaptable human pathogen that might enter a dormant non-replicating (NR), drug-tolerant stage. ...

Abstract	Mycobacterium tuberculosis (Mtb) is the aetiological agent of tuberculosis, the leading cause of death worldwide from a single infectious agent. Mtb is a highly adaptable human pathogen that might enter a dormant non-replicating (NR), drug-tolerant stage. Reactivation of dormant Mtb can lead to active disease. Antibiotic treatments of active and latent tuberculosis are long, complex and may fail to fully eradicate the infection. Therefore, it is imperative to identify novel compounds with new mechanisms of action active against NR bacilli. Dormant Mtb habitat is mostly thought to be the pH-neutral and hypoxic caseous granuloma. We have used the Wayne culture model to reproduce this environment and tested the activities of two DNA-targeted agents, C8-linked-pyrrolobenzodiazepine(PBD)-polyamide conjugates 1 and 2, against Mtb grown in aerobic and hypoxic conditions in both acidic and pH-neutral media. PBD 2 showed growth inhibitory activity at 5.1 µg/ml against 19-day-old hypoxic NR Mtb cultures with 1.8 log
MeSH term(s)	Anaerobiosis ; Antitubercular Agents/pharmacology ; Benzodiazepines/chemistry ; Benzodiazepines/pharmacology ; Cell Line ; Humans ; Isoniazid/pharmacology ; Mycobacterium tuberculosis/drug effects ; Mycobacterium tuberculosis/growth & development ; Nylons/chemistry ; Nylons/pharmacology ; Pyrroles/chemistry ; Pyrroles/pharmacology ; Rifampin/pharmacology ; Tuberculosis, Pulmonary/drug therapy
Chemical Substances	Antitubercular Agents ; Nylons ; Pyrroles ; pyrrolo(2,1-c)(1,4)benzodiazepine ; Benzodiazepines (12794-10-4) ; Isoniazid (V83O1VOZ8L) ; Rifampin (VJT6J7R4TR)
Language	English
Publishing date	2018-05-24
Publishing country	Japan
Document type	Journal Article
ZDB-ID	390800-8
ISSN	1881-1469 ; 0021-8820 ; 0368-3532
ISSN (online)	1881-1469
ISSN	0021-8820 ; 0368-3532
DOI	10.1038/s41429-018-0068-5
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Article ; Online: The Combination Rifampin-Nitazoxanide, but Not Rifampin-Isoniazid-Pyrazinamide-Ethambutol, Kills Dormant Mycobacterium tuberculosis in Hypoxia at Neutral pH.

Iacobino, Angelo / Giannoni, Federico / Pardini, Manuela / Piccaro, Giovanni / Fattorini, Lanfranco

Antimicrobial agents and chemotherapy

2019 Volume 63, Issue 7

Abstract: The activities of rifampin, nitazoxanide, PA-824, and sutezolid were tested against ... ...

Abstract	The activities of rifampin, nitazoxanide, PA-824, and sutezolid were tested against dormant
MeSH term(s)	Antitubercular Agents/pharmacology ; Drug Combinations ; Drug Therapy, Combination ; Ethambutol/pharmacology ; Humans ; Hydrogen-Ion Concentration ; Hypoxia ; Isoniazid/pharmacology ; Microbial Sensitivity Tests ; Mycobacterium tuberculosis/drug effects ; Nitroimidazoles/pharmacology ; Oxazolidinones/pharmacology ; Pyrazinamide/pharmacology ; Rifampin/pharmacology ; Tuberculosis/microbiology
Chemical Substances	Antitubercular Agents ; Drug Combinations ; Nitroimidazoles ; Oxazolidinones ; PNU-100480 ; isoniazid, pyrazinamide, rifampin drug combination ; pretomanid ; Pyrazinamide (2KNI5N06TI) ; Ethambutol (8G167061QZ) ; Isoniazid (V83O1VOZ8L) ; Rifampin (VJT6J7R4TR)
Language	English
Publishing date	2019-06-24
Publishing country	United States
Document type	Journal Article
ZDB-ID	217602-6
ISSN	1098-6596 ; 0066-4804
ISSN (online)	1098-6596
ISSN	0066-4804
DOI	10.1128/AAC.00273-19
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Zs.A 809: Show issues			Location: Je nach Verfügbarkeit (siehe Angabe bei Bestand) bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular Jg. 1995 - 2021: Lesesall (1.OG) ab Jg. 2022: Lesesaal (EG)
Einzelsignatur: Show issues

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Article ; Online: Revisiting problems and solutions to decrease Mycobacterium tuberculosis pyrazinamide false resistance when using the Bactec MGIT 960 system.

Mustazzolu, Alessandro / Piersimoni, Claudio / Iacobino, Angelo / Giannoni, Federico / Chirullo, Barbara / Fattorini, Lanfranco

Annali dell'Istituto superiore di sanita

2019 Volume 55, Issue 1, Page(s) 51–54

Abstract: Pyrazinamide (PZA) is a first-line key drug used in combination with other agents for the treatment of tuberculosis (TB). Phenotypic and molecular assays for testing susceptibility of Mycobacterium tuberculosis (Mtb) to PZA have been developed, with the ... ...

Abstract	Pyrazinamide (PZA) is a first-line key drug used in combination with other agents for the treatment of tuberculosis (TB). Phenotypic and molecular assays for testing susceptibility of Mycobacterium tuberculosis (Mtb) to PZA have been developed, with the assay in liquid medium at acidic pH in the Bactec MGIT 960 (M960) system being routinely used in the mycobacteriology laboratories. However, false resistance to PZA by this method was reported to occur by several investigators, mostly due to high Mtb inoculum, which may impair drug activity by increasing the pH of the medium. In this study, a revision of the literature on the issue of false resistance in the M960 PZA assay was performed. In the reports examined, all improvements of the M960 test proposed to decrease false resistant results were based on the use of reduced inoculum densities of Mtb cells, to be easily translated into laboratory practice.
MeSH term(s)	Antitubercular Agents/pharmacology ; Culture Media/chemistry ; Drug Resistance, Bacterial ; False Positive Reactions ; Humans ; Hydrogen-Ion Concentration ; Microbial Sensitivity Tests/methods ; Mycobacterium Infections/microbiology ; Mycobacterium tuberculosis/drug effects ; Pyrazinamide/pharmacology ; Reproducibility of Results ; Tuberculosis, Multidrug-Resistant/microbiology
Chemical Substances	Antitubercular Agents ; Culture Media ; Pyrazinamide (2KNI5N06TI)
Language	English
Publishing date	2019-02-22
Publishing country	Italy
Document type	Journal Article
ZDB-ID	950344-4
ISSN	2384-8553 ; 0021-2571
ISSN (online)	2384-8553
ISSN	0021-2571
DOI	10.4415/ANN_19_01_09
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Zs.A 231: Show issues

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Article ; Online: Fighting tuberculosis by drugs targeting nonreplicating

Iacobino, Angelo / Piccaro, Giovanni / Giannoni, Federico / Mustazzolu, Alessandro / Fattorini, Lanfranco

International journal of mycobacteriology

2017 Volume 6, Issue 3, Page(s) 213–221

Abstract: Current tuberculosis (TB) treatment requires 6 months of combination therapy with isoniazid (INH), rifampin (RIF), pyrazinamide (PZA), and ethambutol for active TB and 9 months of INH or 3 months of rifapentine (RFP) + INH for latent TB. The lungs of ... ...

Abstract	Current tuberculosis (TB) treatment requires 6 months of combination therapy with isoniazid (INH), rifampin (RIF), pyrazinamide (PZA), and ethambutol for active TB and 9 months of INH or 3 months of rifapentine (RFP) + INH for latent TB. The lungs of patients with active and latent TB contain heterogeneous mixtures of cellular and caseous granulomas harboring Mycobacterium tuberculosis bacilli ranging from actively replicating (AR) to nonreplicating (NR), phenotypically drug-resistant stages. Several in vitro models to obtain NR cells were reported, including exposure to hypoxia, nutrient starvation, acid + nitric oxide, and stationary phase. Overall, these models showed that RIF, RFP, PA-824 (PA), metronidazole (MZ), bedaquiline (BQ), and fluoroquinolones were the most active drugs against NR M. tuberculosis. In hypoxia at pH 5.8, some combinations killed AR plus NR cells, as shown by lack of regrowth in liquid media, whereas in hypoxia at pH 7.3 (the pH of the caseum), only RIF and RFP efficiently killed NR bacilli while several other drugs showed little effect. In conventional mouse models, combinations containing RFP, BQ, PA, PZA, moxifloxacin, sutezolid, linezolid, and clofazimine sterilized animals in ≤2 months, as shown by lack of viable bacilli in lung homogenates after 3 months without therapy. Drugs were less effective in C3HeB/FeJ mice forming caseous granulomas. Overall, in vitro observations and in vivo studies suggest that the search for new TB drugs could be addressed to low lipophilic molecules (e.g., new rpoB inhibitors with clogP < 3) killing NR M. tuberculosis in hypoxia at neutral pH and reaching high rates of unbound drug in the caseum.
Language	English
Publishing date	2017-07
Publishing country	Netherlands
Document type	Journal Article ; Review
ISSN	2212-554X
ISSN (online)	2212-554X
DOI	10.4103/ijmy.ijmy_85_17
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