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  • 1
    Language: English
    In: Journal of bacteriology, November 2012, Vol.194(21), pp.5864-74
    Description: Hfq is an RNA-binding protein known to regulate a variety of cellular processes by interacting with small RNAs (sRNAs) and mRNAs in prokaryotes. Stenotrophomonas maltophilia is an important opportunistic pathogen affecting primarily hospitalized and immunocompromised hosts. We constructed an hfq deletion mutant (Δhfq) of S. maltophilia and compared the behaviors of wild-type and Δhfq S. maltophilia cells in a variety of assays. This revealed that S. maltophilia Hfq plays a role in biofilm formation and cell motility, as well as susceptibility to antimicrobial agents. Moreover, Hfq is crucial for adhesion to bronchial epithelial cells and is required for the replication of S. maltophilia in macrophages. Differential RNA sequencing analysis (dRNA-seq) of RNA isolated from S. maltophilia wild-type and Δhfq strains showed that Hfq regulates the expression of genes encoding flagellar and fimbrial components, transmembrane proteins, and enzymes involved in different metabolic pathways. Moreover, we analyzed the expression of several sRNAs identified by dRNA-seq in wild-type and Δhfq S. maltophilia cells grown in different conditions on Northern blots. The accumulation of two sRNAs was strongly reduced in the absence of Hfq. Furthermore, based on our dRNA-seq analysis we provide a genome-wide map of transcriptional start sites in S. maltophilia.
    Keywords: Host Factor 1 Protein -- Metabolism ; Molecular Chaperones -- Metabolism ; RNA, Bacterial -- Metabolism ; Stenotrophomonas Maltophilia -- Physiology
    ISSN: 00219193
    E-ISSN: 1098-5530
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  • 2
    Language: English
    In: The Journal of Pediatrics, 2009, Vol.154(6), pp.854-858
    Description: To determine the prevalence of and explore possible differences in the risk for and symptoms of infection between patients with and without inflammatory bowel disease (IBD). Stool specimens from subjects with and without IBD were evaluated for the presence of toxins. Demographic information, diagnosis, anatomic location, disease activity, IBD therapy, hospitalizations, and antibiotic and proton pump inhibitor (PPI) exposures were recorded. A total of 193 specimens were collected from 81 patients with IBD and 112 patients without IBD. The prevalence of infection was significantly greater in the patients with IBD than in those without IBD ( = .004; χ = 0.003; odds ratio = 3.3; 95% confidence interval = 1.5 to 7.6). In the patients with IBD, the prevalence of active disease was significantly greater in the --infected patients than in the uninfected patients ( 〈 .0001). Colonic involvement was found in all patients with IBD. The specific type of IBD, IBD therapy, and antibiotic and PPI exposures that predisposed patients with IBD to infection were not identified, whereas hospitalization was significantly more frequent in the patients without IBD ( = .025). Our findings indicate that in children, IBD is associated with an increased prevalence of infection. The specific risk factors reported in adults were not identified in these children, suggesting the possible involvement of other mechanisms for acquiring the pathogen.
    Keywords: Medicine
    ISSN: 0022-3476
    E-ISSN: 1097-6833
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  • 3
    Language: English
    In: Scientific reports, 04 December 2018, Vol.8(1), pp.17570
    Description: Candida species cause cutaneous and systemic infections with a high mortality rate, especially in immunocompromised patients. The emergence of resistance to the most common antifungal drugs, also due to biofilm formation, requires the development of alternative antifungal agents. The antimicrobial peptide VLL-28, isolated from an archaeal transcription factor, shows comparable antifungal activity against 10 clinical isolates of Candida spp. Using a fluoresceinated derivative of this peptide, we found that VLL-28 binds to the surface of planktonic cells. This observation suggested that it could exert its antifungal activity by damaging the cell wall. In addition, analyses performed on biofilms via confocal microscopy revealed that VLL-28 is differentially active on all the strains tested, with C. albicans and C. parapsilosis being the most sensitive ones. Notably, VLL-28 is the first example of an archaeal antimicrobial peptide that is active towards Candida spp. Thus, this points to archaeal microorganisms as a possible reservoir of novel antifungal agents.
    Keywords: Antifungal Agents -- Pharmacology ; Antimicrobial Cationic Peptides -- Pharmacology ; Archaeal Proteins -- Pharmacology ; Biofilms -- Drug Effects ; Candida -- Drug Effects
    E-ISSN: 2045-2322
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  • 4
    Language: English
    In: BMC genomics, 14 November 2015, Vol.16, pp.933
    Description: A giant protein called BAP (biofilm-associated protein) plays a role in biofilm formation and adhesion to host cells in A. baumannii. Most of the protein is made by arrays of 80-110 aa modules featuring immunoglobulin-like (Ig-like) motifs. The survey of 541 A. baumannii sequenced strains belonging to 108 STs (sequence types) revealed that BAP is highly polymorphic, distinguishable in three main types for changes both in the repetitive and the COOH region. Analyzing the different STs, we found that 29 % feature type-1, 40 % type-2 BAP, 11 % type-3 BAP, 20 % lack BAP. The type-3 variant is restricted to A. baumannii, type-1 and type-2 BAP have been identified also in other species of the Acinetobacter calcoaceticus-baumannii (ACB) complex. A. calcoaceticus and A. pittii also encode BAP-like proteins in which Ig-like repeats are replaced by long tracts of alternating serine and aspartic acid residues. We have identified in species of the ACB complex two additional proteins, BLP1 and BLP2 (BAP-like proteins 1 and 2) which feature Ig-like repeats, share with BAP a sequence motif at the NH2 terminus, and are similarly expressed in stationary growth phase. The knock-out of either BLP1 or BLP2 genes of the A. baumannii ST1 AYE strain severely affected biofilm formation, as measured by comparing biofilm biomass and thickness, and adherence to epithelial cells. BLP1 is missing in the majority of type-3 BAP strains. BLP2 is largely conserved, but is frequently missing in BAP-negative cells. Multiple proteins sharing Ig-like repeats seem to be involved in biofilm formation. The uneven distribution of the different BAP types, BLP1, and BLP2 is highly indicative that alternative protein complexes involved in biofilm formation are assembled in different A. baumannii strains.
    Keywords: Acinetobacter Baumannii -- Genetics ; Bacterial Proteins -- Genetics
    E-ISSN: 1471-2164
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  • 5
    Language: English
    In: International Journal of Molecular Sciences, 01 August 2015, Vol.16(9), pp.20375-20391
    Description: Photofrin/photodynamic therapy (PDT) at sub-lethal doses induced a transient stall in proteasome activity in surviving A549 (p53+/+) and H1299 (p53−/−) cells as indicated by the time-dependent decline/recovery of chymotrypsin-like activity. Indeed, within 3 h of incubation, Photofrin invaded the cytoplasm and localized preferentially within the mitochondria. Its light activation determined a decrease in mitochondrial membrane potential and a reversible arrest in proteasomal activity. A similar result is obtained by treating cells with Antimycin and Rotenone, indicating, as a common denominator of this effect, the ATP decrease. Both inhibitors, however, were more toxic to cells as the recovery of proteasomal activity was incomplete. We evaluated whether combining PDT (which is a treatment for killing tumor cells, per se, and inducing proteasome arrest in the surviving ones) with Bortezomib doses capable of sustaining the stall would protract the arrest with sufficient time to induce apoptosis in remaining cells. The evaluation of the mitochondrial membrane depolarization, residual proteasome and mitochondrial enzymatic activities, colony-forming capabilities, and changes in protein expression profiles in A549 and H1299 cells under a combined therapeutic regimen gave results consistent with our hypothesis.
    Keywords: Photodynamic Therapy ; Photofrin ; Bortezomib ; Combination Therapy ; Proteasome ; Biology
    E-ISSN: 1422-0067
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  • 6
    In: APMIS, January 2014, Vol.122(1), pp.42-46
    Description: Strains of producing ‐carbapenemase have emerged as one of the most important multidrug‐resistant Gram‐negative nosocomial pathogens. Here, we report the first isolation and subsequent dissemination of a 512 producing ‐3 carbapenemase in a hospital in southern Italy. Isolates were obtained from blood, throat swabs, sputum, catheters, and urine of patients admitted to different hospital wards. Antimicrobial s were determined for all isolates by automated systems and confirmed by Etest. Carbapenemase production was confirmed by the modified Hodge test and by a disc synergy test, and carbapenemase genes were investigated by . All isolates were characterized by pulse‐field gel electrophoresis () and multilocus sequence typing () analysis. Most isolates were multidrug resistant with exception of some isolates intermediately susceptible to gentamicin, tigecycline, and trimethoprim‐sulfamethoxazole. analysis showed that isolates harbored the bla gene associated with bla and bla. and showed that all isolates belonged to the same 512 clone recently described in Israel.
    Keywords: Klebsiella Pneumoniae 512 ; Bla ; Multidrug Resistance ; Outbreak ; Bla
    ISSN: 0903-4641
    E-ISSN: 1600-0463
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  • 7
    Language: English
    In: International Journal of Molecular Sciences, 01 August 2015, Vol.16(9), pp.20417-20430
    Description: Although photodynamic therapy (PDT), a therapeutic approach that involves a photosensitizer, light and O2, has been principally considered for the treatment of specific types of cancers, other applications exist, including the treatment of infections. Unfortunately, PDT does not always guarantee full success since it exerts lethal effects only in cells that have taken up a sufficient amount of photosensitizer and have been exposed to adequate light doses, conditions that are not always achieved. Based on our previous experience on the combination PDT/chemotherapy, we have explored the possibility of fighting bacteria that commonly crowd infected surfaces by combining PDT with an antibiotic, which normally does not harm the strain at low concentrations. To this purpose, we employed 5-aminolevulinic acid (5-ALA), a pro-drug that, once absorbed by proliferating bacteria, is converted into the natural photosensitizer Protoporphyrin IX (PpIX), followed by Gentamicin. Photoactivation generates reactive oxygen species (ROS) which damage or kill the cell, while Gentamicin, even at low doses, ends the work. Our experiments, in combination, have been highly successful against biofilms produced by several Gram positive bacteria (i.e., Staphylococcus aureus, Staphylococcus epidermidis, etc.). This original approach points to potentially new and wide applications in the therapy of infections of superficial wounds and sores.
    Keywords: Photodynamic Therapy ; 5-Aminolevulinic Acid ; Gentamicin ; Combination Therapy ; Biology
    E-ISSN: 1422-0067
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  • 8
    Language: English
    In: Frontiers in microbiology, 2015, Vol.6, pp.723
    Description: Stenotrophomonas maltophilia is increasingly identified as an opportunistic pathogen in immunocompromised, cancer and cystic fibrosis (CF) patients. Knowledge on innate immune responses to S. maltophilia and its potential modulation is poor. The present work investigated the ability of 12 clinical S. maltophilia strains (five from CF patients, seven from non-CF patients) and one environmental strain to survive inside human monocyte-derived dendritic cells (DCs). The effects of the bacteria on maturation of and cytokine secretion by DCs were also measured. S. maltophilia strains presented a high degree of heterogeneity in internalization and intracellular replication efficiencies as well as in the ability of S. maltophilia to interfere with normal DCs maturation. By contrast, all S. maltophilia strains were able to activate DCs, as measured by increase in the expression of surface maturation markers and proinflammatory cytokines secretion.
    Keywords: DC Maturation Markers ; Cystic Fibrosis ; Cytokines Secretion ; Innate Immune Response ; Opportunistic Pathogen
    ISSN: 1664-302X
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  • 9
    In: APMIS, June 2016, Vol.124(6), pp.516-521
    Description: has recently emerged as an important hospital pathogen. In this study, we showed the emergence of isolates carrying the gene in patients colonized by carbapenem‐resistant strains. Two multiresistant isolates were recovered from bronchial aspirates of two patients hospitalized in the Intensive Care Unit at the “Santa Maria della Scaletta” Hospital, Imola. The antimicrobial susceptibility test showed the high resistance to carbapenems and double‐disk synergy test confirmed the phenotype of and AmpC production. Other investigation revealed that and genes were carried on the conjugative plasmid. This is a relevant report in Italy that describes a nosocomial infection due to the production of beta‐lactamases by an isolate in patients previously colonized by carbapenem‐resistant. In conclusion, it's necessary a continuous monitoring of multidrug‐resistant strains for the detection of any ‐producing bacteria that could expand the circulation of carbapenem‐resistant pathogens.
    Keywords: Kpc Gene ; Carbapenem Resistence ; Nosocomial Infection ; Surveillance
    ISSN: 0903-4641
    E-ISSN: 1600-0463
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  • 10
    Language: English
    In: The new microbiologica, April 2015, Vol.38(2), pp.251-7
    Description: Acinetobacter baumannii is a multidrug-resistant pathogen associated with severe infections in hospitalized patients, including pneumonia, urinary and bloodstream infections. Rapid detection of A. baumannii infection is crucial for timely treatment of septicemic patients. The aim of the present study was to develop a specific marker for a quantitative polymerase chain reaction (PCR) assay for the detection of A. baumannii. The target gene chosen is the biofilm-associated protein (bap) gene, encoding a cell surface protein involved in biofilm formation. The assay is specific for A. baumannii, allowing its discrimination from different species of Acinetobacter and other clinically relevant bacterial pathogens. The assay is able to detect one genomic copy of A. baumannii, corresponding to 4 fg of purified DNA, and 20 colony-forming units/ml using DNA extracted from spiked whole blood samples.
    Keywords: Bacterial Bloodstream Infection ; Blood Culture ; Molecular Diagnostics ; Sepsis ; Taqman Real-Time Pcr ; Acinetobacter Infections -- Microbiology ; Acinetobacter Baumannii -- Isolation & Purification ; Blood -- Microbiology ; Real-Time Polymerase Chain Reaction -- Methods
    ISSN: 1121-7138
    Source: MEDLINE/PubMed (U.S. National Library of Medicine)
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